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USER MANUAL

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1. Enter the second wavelength as above and repeat for the a number of wavelengths selected up to 5 Step 3 Press OK O to enter the results screen oox eN Press Cancel to return to the Applications Folder Multi wavelength Step 4 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 5 Insert sample and press DJ Repeat step 5 for all samples Results A scan plot covering the range of wavelengths selected with cursors at the relevant wavelengths and a table of values is displayed Press to return to the Applications Folder Press lt gt to display available Options which are described below Parameters Print Print Graph Save Method Auto Print We O o GO E sample Mumber o Version 5 4 18 4 2011 Page 28 Picodrop Ltd 7 Absorbance Ratio This makes simple absorbance ratio measurements on samples measuring the amount of light that has passed through a sample relative to a blank this can be air at two wavelengths The procedure is as follows Step 1 Absorbance Ratio Wavelengths Enter the first wavelength by using the keypad numbers or the left and right arrows wavelength 1 Press the down arrow Step 2 Enter the second wavelength as above wavelength 7 Press the down arrow 280 nm Step 3 Select whether a background correction is applied to both A wavelengths 1 and 2 using the left and right arrows Step 4 If bac
2. If the instrument with this accessory fitted is used in a manner not specified or in environmental conditions not appropriate for safe operation then the protection provided may be impaired and instrument warranty withdrawn There are no user serviceable parts inside this accessory Version 5 4 18 4 2011 Page 68 Picodrop Ltd Fitting SD memory card accessory Step 1 Remove the power cable from the instrument Turn the instrument over and place onto a soft surface for example a folded up towel remove cap head screws from the positions indicated using the Allen key provided Step 2 Turn the instrument back over and lift the accessory cover vertically upwards to remove Remove the tie wrap from the cable Step 3 Plug the accessory cable into the SD memory card PCB module Step 4 Note the slots in the base of the case The two lugs on the SD memory card PCB module plug into these d Version 5 4 18 4 2011 Page 69 Picodrop Ltd Step 5 Note the slots in the accessory cover supplied these are designed to engage with the SD memory card PCB module Step 6 Lower the accessory cover vertically downwards onto the instrument engaging the PCB in the slots Step 7 Invert the instrument and replace the cap head screws using the Allen key provided The accessory is now ready for use Version 5 4 18 4 2011 Page 70 Picodrop Ltd OPERATION SD memory cards are inserted into the accessory with th
3. Do not proof test Strength or proof testing is done by the fibre manufacturers It is a carefully controlled process because it has been found that the more one stresses fibre near its strength limit reduces it overall life time expectation to continue to resist failures Therefore it is specifically recommended that no fibre be proof tested or strength tested except for sample testing in order to gather nominal strength statistics Any overall proof or strength testing is one at the risk of damaging the fibre Specifically proof testing a fibre at or below its bend radius minimum or in any way uncontrolled or unapproved by Picodrop Ltd will void the warranty Bend Radius Do not bend the fibre past the minimum bend radius The rule of thumb regarding bend radius is that the Minimum bending radius for a particular fibre is 300x the cladding diameter assuming the cladding is also glass Twisting Do not coil or twist the cable when spooling un spooling coiling or uncoiling Cables must be handled ina hand over hand fashion at all times Fibre cables should not be handled like rope and coiled or uncoiled by twisting or untwisting one loop at a time Fibre should always be handled in a hand over hand fashion making coils or unrolling coils by moving the fibre in a circle one hand over the other To do otherwise is to induce a twisting stress in the cable and hence in the fibre within the cable Very high stresses can be achie
4. WEEE 2002 95 EC ROHS directive Standards to which conformity is declared where relevant are as follows EN61010 1 2010 Safety requirements for electrical equipment for measurement control and laboratory use General requirements EN61326 1 2006 Electrical equipment for measurement control and laboratory use EMC Requirements EN 12100 1 2 2003 A1 2009 Safety of machinery Basic concepts general principles for design EN 14121 1 2007 Safety of machinery Risk assessment P200BT and P200BTE also conform to the requirements of the following Directives 1999 5 EC Radio and Telecommunications terminal equipment directive for Bluetooth devices This equipment has been tested and found to comply with the limits fora CLASS A digital device pursuant to part 15 of the FCC Rules Version 5 4 18 4 2011 Page 4 Picodrop Ltd ESSENTIAL SAFETY NOTES Caution symbol There are a number of warning labels and symbols on your instrument These are there to inform you where potential danger exists or particular caution is required Before commencing installation please take time to familiarise yourself with these symbols and their meaning There are no bio hazardous materials within the unit however this unit could be used with bio hazardous samples Before using the instrument the customer should have in place decontamination procedures designed to protect laboratory workers from occupationally acquired infections In the event of a spillage
5. back into the same instrument at a later date There is the option to Backup Load either a single folder of methods or to Backup Load all folders on the instrument To backup the METHOD Folder go to methods and press OPTIONS followed by SD memory Card r kas w ie hth ee ths E Stns O W de R as as as Methods 1 Methods 2 Methods 3 a ce en ce EA ons Hite ect by pe Eii by pe ae he Methods 4 Methods E Methods E Folder Hames E PA Lock Folder E a E Unlock Folder iy SO Memor Card Methods 3 Operation q Backup All Folders GA Cancel E BB Cancel Alternatively inserting an SD memory card whilst the method screen is displayed will also bring up the same screen Folder or folders will then be written to the SD memory Card the card must not be removed when the LED is lit otherwise the Methods on the card will be corrupted Methods are stored on the SD memory card in the Serial no BACKUP Directory All folder names are also stored on the card Different instruments can be backed up to the same card as methods will be stored under different sub directories due to the different instrument serial numbers Version 5 4 18 4 2011 Page 74 Picodrop Ltd Restoring method folders This allows a previous backup of a method folder or all method folders to be restored to the original instrument Methods Methods Operation Operation hi Restore Folder Z g 4 Restore All Folders d ml Folder F
6. the last digit entered Step 10 Press Next to enter the Calibration screen If there are duplicate or non monotonic increasing entries the unit will beep and highlight the incorrect entry OR Press Back to return to the Parameter screen Page 44 Picodrop Ltd Calibration Screen replicates off This shows the calibration values and allows standards to be Bradford Calibration measured Step 11 3 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result A ar ara Repeat step 12 for all standards A graph will display the results and the fitted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 When all standards are measured the OK box appears Press Bradford Calibration to accept the calibration an go to the Results screen see Standards below 0 70 0 058 A O R 0 40 0 1965 F E 0 60 0 3304 A Press Back to cancel selections and return to the Standards 0 80 0 4634 04 i screen 1 00 0 5392 4 i 1 40 0 7194 na e Calibration Screen replicates on y This shows the calibration values and allows standards to be oe 04 05 08 Lo La 14 measured risk Insert the reference Press EE key Thi
7. Absorbance Ratio Insert sample and press DJ Repeat step 11 for all samples The absorbance at selected wavelengths is measured and the ratio between wavelengths 1 and 2 is calculated both corrected by the background wavelength value if this was selected Concentration 0 26 pal ml Press to return to the Applications Folder Press C Options d to display available Options which are described below Options select using key pad numbers Return to parameters screen step 1 above 2 Print result via selected method UD Parameters 3 Toggle graph on off Graph shows a wavescan plot across Print the selected wavelengths in place of the individual gah wavelength 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder E Sample Number to store in Favourites Methods 1 9 press the down arrow E Save Method and enter name E Auto Print we 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 30 Picodrop Ltd THE LIFE SCIENCE FOLDER Life Science Folder This contains three sub folders Nucleic Acids Protein and Cell Count Contents of these sub folders are detailed below 1 Nucleic Acids Concentration and purity check for DNA samples SCA RNA Ci SCSCSCSCSCSCSCSCC Concentration and purity check for RNA samples 1
8. FOLDER Utilities o O ate and Time Regional Printer o0 eof Contrast Folder Mames Summary Function Keypad number Description de 1 Set correct time and date Date and Time 2 Select preferred language and number format Regional 3 Printer output options Frinter 4 Select screen layout themes and history Preferences 5 Adjust screen contrast amp brightness Contrast O al 6 Re name Method folders Folder Hames i 7 Serial number and software version o al 8 Spectro Blocks Sudoku Version 5 4 18 4 2011 Page 56 Picodrop Ltd Utilities 1 Date and Time The procedure is as follows 2 Regional Enter the day using the keypad numbers or left and right arrows Press the down arrow Enter the month as above Day Hour Press the down arrow Enter the year Press the down arrow va a Enter the hour A Enter the minute Seconds are zeroed when OK is pressed TER Press OK to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the time Date and Time Sets Language and Number Format The procedure is as follows Regional Select a language Options are French English or Spanish Language German and Italian will be released in the near future Press the down arrow Set the decimal point style Options are or Seer e Press OK to store the settings and return to the Utilities folder Press Cance
9. Page 71 Picodrop Ltd When a method is being stored the LED next to the card will light up the card MUST not be removed whilst the light is on otherwise the stored method will be corrupted Loading methods from SD memory card Selecting the SD memory card by pressing the relevant number on the home page shows the methods stored on the card SD Memory Card ofA eli eff Single wavelength Concentration Wi aVescan of ef eff Wr aWescang Wr aWescanad Wi aWVescanadg ei ef eff We aWescanadgm Wi avescanadgmag VW avescanadgmam The required method can be loaded by pressing the relevant number on the keyboard and run in the same way as methods stored in any of the method folders on the instrument Saving data to SD memory card Data from all applications on your instrument can be stored onto the SD memory card To enable data to be stored on the card the SD memory card must be selected as the output device to do this select utilities printer and under printer select SD memory card and ensure Auto Print is selected Printer Auto Print On Printer 4 SD Memory Card P Version 5 4 18 4 2011 Page 72 Picodrop Ltd Start an application or load a method in the usual way Note that when SD memory card and Auto Print are both selected in utilities an SD icon appears in the top right hand corner of the display Concentration A260 4 230 For applications that print continuously such as Single Wavelength
10. Return to parameters screen step 1 above 2 Print result via selected method EP Parameters 3 Toggle graph on off Displays the calibration graph cursors E Frin give values for last measured sample Y Graph 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow EA Sample Number and enter name EP save Method 9 Auto print toggles auto print on off Auto Print Exit options by pressing or wait Version 5 4 18 4 2011 Page 43 Picodrop Ltd 3 Bradford The procedure is as follows Bradford Parameters wavelength 535 nm Standards Bradford Parameters wavelength 555 nm Standards Units wat mol Bradford Standards Std 2 0 40 pas rl Std 3 0 60 pa rl Version 5 4 18 4 2011 Curve Fit Regression Calibration Standards Replicates Curve Fit Calibration Replicates Std 4 4 20 was mil Std 5 1 00 pas rl Std 6 1 40 pa mil Step 1 Press 3 to select Bradford method Step 2 Wavelength for this stored method is pre set to 595 nm Step 3 Enter the number of standard concentration points 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 Units The user can enter a text string up to 8 characte
11. Slope Az result calculated from the selected parameter dA final A or oom 0 006 0 005 oro om oxsss Slope Sample 1 Time 00 00 10 Abs 0 231 o ba Use the left and right arrows to move the cursor and display the time and absorbance value at measured data points Use the up and down arrows to zoom in or out Press Cancel to return to the Applications Folder Press lt gt to display available Options which are described below Version 5 4 18 4 2011 Page 22 Picodrop Ltd 1 Parameters 3 Print E Print Data C Set tO At Cursor E Set At Cursor Options select using key pad numbers O slope 1 Return to parameter 1 screen step 1 above E Sample Number 2 Print data on the results screen via selected method E Save Wlethod 3 Print all the data O suto Print A 4 Set the t position starting point for the slope and dA calculation at the current cursor position Value is retained for subsequent samples 5 Set the t position finishing point for the slope and dA calculation at the current cursor position Value is retained for subsequent samples 6 Toggle the calculated slope line on and off Note if any data points enclosed by t and t are beyond the range of the instrument gt 2 5A or lt 0 3A then this option is greyed out 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use t
12. Step 3 calculate dilution factor Press to enter the dilution factor screen Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press O to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Select the units of measurement using the left and right arrows Options ug ml ng ul ug ul and pmol ul If pmol ul is selected the factor changes to a selection table denoting the ratios of the 4 bases in the structure Press the down arrow Step 6 units not pmol l a Enter the factor using the keypad numbers Default value is 33 mm range is 0 01 to 9999 OR Dilution Factor Step 6 units pmol ul ee Enter the proportions of bases present using the keypad M Oligo Parameters numbers and up and down arrows to move between boxes Background Default is 10 for each range is 0 to 9999 Step 7 Press OK D to enter the Results screen OR Cancel to return to the Nucleic Acids folder Version 5 4 18 4 2011 Page 36 Picodrop Ltd Results Screen Step 8 Insert the reference Press GE Key This will be used for all subsequent samples until changed Step 9 Insert sample and
13. Step 9 Lowry Standards ees E Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears E o the last digit entered Alepo Press Next to enter the Calibration screen If there are duplicate or non monotonic increasing entries the unit will beep and highlight the incorrect entry OR Press Back to return to the Parameter screen Version 5 4 18 4 2011 Page 47 Picodrop Ltd Calibration Screen replicates off This shows the calibration values and allows standards to be Lowry Calibration measured s Pp Toa Insert the reference Press GEM key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result TH 0 4 0 6 0 8 1 0 L 2 1 34 i a Repeat step 12 for all standards A graph will display the results and the fitted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 Lowry Calibration When all standards are measured the OK box appears Press Standards to accept the calibration an go to the Results screen see 0 0174 cd below 0 0564 mas af OR 0 095 A 0 1324 oni 0 1694 0 205 4 ru Press Back to c
14. Within each application the user has the possibility to select various options that define the way results are treated If not using a stored method it is advisable to check that these Options have been appropriately set for your experiment when coming to the instrument Note that setting the History parameter to on see Preferences later will cause the instrument to store its last settings If the History parameter is turned off all parameters and options will return to their default settings when you leave that application Unless it has been saved as a method Version 5 4 18 4 2011 Page 13 Picodrop Ltd 1 Single Wavelength Abs and T This makes simple absorbance A and transmission T measurements on samples measuring the amount of light that has passed through a sample relative to a reference this can be air The procedure is as follows Single Wavelength Parameters Step 1 Set wavelength by using keypad numbers or left and right wavelength arrows Press the down arrow key Step 2 Select the mode Absorbance or T using the left and right arrows Step 3 To enter the results screen with the selected parameters press OK OR Cancel the selections and return to the Applications Folder by pressing Cancel Single wavelength Step 4 Insert the reference Press GE key This will be used for all subsequent samples until changed Step 5 Insert sample and press DJ O Repeat step 5 for
15. all samples Single wavelength Results The result at the selected wavelength is displayed on screen Use the left and right arrows to move the cursor and display the value at the cursor position 15nm from set wavelength Press Cancel to return to the Applications Folder Press ME to display available Options which are described below Version 5 4 18 4 2011 Page 14 Picodrop Ltd Options select using key pad numbers d Parameters O Fin 1 Return to parameters screen step 1 above O 2o T 2 Print result via selected method GB Print Graph 3 Toggle between Absorbance and I mode 4 Print graph greyed out if no data are available 7 Sample number add a prefix to the sample number and gt aes reset the incrementing number to the desired value Save Method 8 Save method use the left and right arrows to select a folder EE autori o vi to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 15 Picodrop Ltd 2 Concentration This makes simple concentration measurements on samples by measuring the amount of light that has passed through a sample relative to a reference this can be air Concentration is obtained by multiplying the measured absorbance at a specific wavelength by a factor The factor may be known in advance or may be calculated by the instrument
16. enter a text string up to 8 characters long To access a list of pre defined units press the Options key and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters or Cancel Y Kinetics Parameters 2 Mode Step 8 Set the Factor by which the result is multiplied to give the amount in the chosen range using the left and right arrows Range of 0 01 to 9999 Factor Step 9 Press Next O to enter the Results screen E OK E Back OR Press Cancel to return to the Parameters 1 screen Results Insert the reference and press the 0A 100 T key Kinetics i A Insert the sample and press to start the run Time min is displayed at the bottom of the screen and A absorbance data are plotted on the graph as testing proceeds A The table below the graph gives absorbance values at T start ad o of calculation T finish of calculation change in absorbance a a 0 6 slope regression parameter RS of the calculated slope and the 0 0 0 4 08 10 Ag An dA
17. parameters or Cancel Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin interpolated or cubic spline Press the down arrow Step 6 Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates using the left and right arrows This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2 or 3 Step 8 Press Next O to enter the Standards screen OR Press Cancel to cancel selections and return to the Protein folder Standards Screen Step 9 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears the last digit entered Step 10 Press Next to enter the Calibration screen If there are duplicate or non monotonic increasing entries the unit will beep and highlight the incorrect entry OR Press Back to return to the Parameter screen Page 41 Picodrop Ltd BCA Calibration 2 5 i la 0 4 0 6 08 Lo l2 1 4 BCA Calibration Standards 0 057 A 0 177 A 0 238 A 0 404 4 05164 A 0 527A B y qe n 0 4 0 6 08 Lo Le 14 oja m Ad a o mu h 4 4 qe o oO BCA Calibration Replicates 1 0 057 A Fl 0 057
18. pressing Cancel Step 5 Insert the reference Press GE key This will be used for all subsequent samples until changed Step 6 Insert sample and press Repeat step 6 for all samples Results A graph of the wavescan is displayed along with a table of Absorbance T at each peak Use the left and right arrows to move the cursor along the graph When it reaches a peak the peak height and width of the peak is displayed at the top of the screen To zoom in on the wavelength scale use the up arrow This auto scales on the Absorbance T scale dependent on the Graph Scale option and this is retained for subsequent measurements To zoom out again use the down arrow Press to return to the Applications Folder Press lt gt to display available Options which are described next Page 18 Picodrop Ltd Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method Ud Parameters 3 Toggle between Absorbance and T mode S Print 4 Displays Peak Detection Parameter Screen See description Abs T below Peak Detection 5 Manually adds a peak position to the peak table in the GP Add Peak results screen at the position set by the cursor If the cursor P Graph Scale is returned to this position the legend User Defined Peak is EY sample Number displayed at the top of the scan and this option changes to E Save Method Delete Peak E sute Pri
19. that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters or Cancel Step 4 Select the type of curve fit using the left and right arrows Options straight line regression a zero regression this forces Standard Curve Parameters A Curve Fit the straight line through the origin interpolated or cubic spline 430 nm 4 Zero Regression PF Step 5 Select the calibration mode either Standards measure prepared Standards Calibration standards or Manual keypad data entry Press the down arrow Step 6 if standards has been selected in step 5 Replicates Select the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2 or 3 Step 7 Press Next to enter the Standards screen OR Press Cancel to cancel selections and return to the Applications Folder Version 5 4 18 4 2011 Page 24 Picodrop Ltd Standard Curve Standards Standards screen Step 8 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 Step 9 Press Next to enter the Calibration screen If any duplicate or non monotonic increasing entries are present the unit will beep and highlight the incorrect entry OR Press Back to return to the Par
20. the area should be disinfected rinsed with distilled water and then allowed to dry The exterior may be wiped with a suitable disinfectant cleaning wipe e Decontamination Equipment returned for repair should include an appropriate decontamination certificate e Itis the responsibility of the customer to ensure that the user of the equipment is trained in the techniques used and is provided with a safe working environment e Any chemicals used in Analyses should be used stored and disposed of in accordance with manufacturer s guidelines and local safety regulations e Toxic Fumes Efficient laboratory ventilation must be provided when working with volatile solvents or toxic substances e Waste disposal Disposal of some solvents and chemicals may be classed as hazardous waste and must be dealt with in accordance with local regulatory practice e Personal protective equipment This is not required to operate the unit but the samples measured may require PPE A local risk assessment should be carried out The lamp source within the unit is flashed to produce UV and visible spectral energy through the sample The energy level is low confined to the optical path and not a hazard in normal use Do not operate the unit with the optical fibre connections disconnected as prolonged exposure to the beam could be hazardous to eyes and skin Unpacking Positioning and Installation e Check the contents of the pack against the packing list If any shorta
21. to clear the previous reading Step 13 When all standards are measured the OK box appears Press Biuret Calibration to accept the calibration an go to the Results screen see below OR Press Back to cancel selections and return to the Standards screen Calibration Screen replicates on This shows the calibration values and allows standards to be measured Step 11 Insert the reference Press EE key This will be used for all subsequent samples until changed Step 12 n 0 4 D G 0 8 1o lg 1 4 Press to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press to measure the standard and Biuret Calibration store the result Standards 0 044 4 a 0 1484 0 250 A 0 349 A 0 446 4 0 542 A Repeat for all replicates and standards j 4 4 ba a A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading cit E pio o mia e sjaj la n 0 4 DE DE Lo l2 1 4 Step 13 D Back EEC Press to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Version 5 4 18 4 2011 Page 51 Picodrop Ltd Biuret Calibration Standards 0 20 0 40 0 60 0 30 1 00 1 40 0 044 4 0 148 A 0 249 A 0 349 4 0
22. validation checks be carried out using traceable standards and that when measuring batches of samples that known standards be included for comparison and verification of result values e Please read through this user manual prior to use e Please contact your original supplier in the first instance if you experience technical or sample handling difficulties If this equipment is used in a manner not specified or in environmental conditions not appropriate for safe operation the protection provided by the equipment may be impaired and instrument warranty withdrawn WEEE The crossed out wheeled bin symbol indicates that the product is covered by the Waste Electrical and Electronic Equipment WEEE Directive and is not to be disposed of as unsorted municipal waste Any products marked with this symbol must be collected separately and in accordance with local regulatory practice Version 5 4 18 4 2011 Page 5 Picodrop Ltd INTRODUCTION Your spectrophotometer Your spectrophotometer is a simple to use UV Visible instrument with a CCD array detector 1024 pixels It has no moving parts which is the basis of the rapid scanning operating system The Picodrop is a microlitre spectrophotometer that provides the user with the facility to recover their sample after measurements have been taken It is a full spectrum 220 950nm spectrophotometer which allows for measurements of common laboratory samples such as DNA RNA and protein in small
23. 005 Abs or 1 of the reading whichever is the greater 546 nm Photometric reproducibility 0 003 Abs 0 to 0 5 Abs 0 007 Abs 0 5 1 0 Abs Stray light lt 0 5 at 220 nm and 340 nm using NaNOz Zero Stability 0 01 Abs hour after 20 min warm up 340 nm Noise Digital output Dimensions Weight Power input 0 005 pk to pk 0 002 RMS USB port standard Bluetooth option 260 x 390 x 100 mm lt 4 5 kg 90 250 V 50 60 Hz Max 30 VA Specifications are measured after the instrument has warmed up at a constant ambient temperature and are typical of a production unit As part of our policy of continuous development we reserve the right to alter specifications without notice Version 5 4 18 4 2011 Page 79 Picodrop Ltd WARRANTY e Your supplier guarantees that the product supplied has been thoroughly tested to ensure that it meets its published specification The warranty included in the conditions of supply is valid for 12 months only if the product has been used according to the instructions supplied The supplier can accept no liability for loss or damage however caused arising from the faulty or incorrect use of this product Fibre Optic Cable Handing and Limits of Warranty Scope The intent of this note is to define the recommended handling practices for fibre optic cables supplied by Picodrop Ltd and to comment on warranty issues Section 1 Handling Guidelines A B C D E Strength and Proof Testing
24. 1 1 Summary Function Keypad number Description o 1 Single wavelength Concentration Wavelength scan Kinetics Standard Applications Curve Multiple wavelengths and Ratio a 2 Saved User selected and configured methods Favourites 3 Sub folder selection for User selected and configured methods Methods o 4 Instrument set up date time language etc and Games Utilities o NT 5 Nucleic acids Proteins and Cell counting Life Science Version 5 4 18 4 2011 Page 12 Picodrop Ltd THE APPLICATIONS FOLDER Applications of om Single wavelength Concentration Ma avescamn oO eB o Kinetics Standard Cure Multi wavelength A o Absorbance Ratio SUMMARY Function Key pad number Description of al 1 Absorbance or T transmission at a single user defined Single wavelength wavelength N e Concentration measurement at a single wavelength based on a E simple Factor entered or calculated from a single standard o 3 Wavelength scan between two user defined wavelengths Range Wiavescan 220 950 nm with user configurable peak finding function oO 2 4 Absorbance versus time measurements either rate or end value Kinetics based o 5 Generation of calibration curve by measuring standards at a single E wavelength tandar ore 6 Absorbance or T transmission at up to 5 user defined A o wavelengths Multi wavelength Pla non O 7 Ratio of absorbance values at two user specified wavelengths Absorbance Ratio Options
25. 3 Oligo Concentration and purity check for oligo samples E A AAA AS E AS UV protein Christian Warburg Protein determination at 280nm ES po Bradford i Protein determination at 595nm po 4 Lowry i Protein determination at 750nm 22222222 5 Buret__ Protein determination a Stom o ao A ee AAA EZ 3 Cell Count OD600 Cell culture OD600 with correction factor EN A CAN DNA RNA and oligonucleotide characterization Nucleic Acid Quantification NAQ e Nucleic acids can be quantified at 260 nm because it is well established that a solution of DNA in a 10 mm pathlength cell with an optical density of 1 0 has a concentration of 50 ug ml or 40 ug ml in the case of RNA Oligonucleotides have a corresponding factor of 33 ug ml although this does vary with base composition this can be calculated if the base sequence is known Concentration Abs260 Factor e The instrument uses factors 50 40 and 33 as defaults for DNA RNA and oligonucleotides respectively and compensates for dilution and use of cells which do not have 10 mm pathlength dilution factor and cell tip pathlength can be entered Nucleic Acid Purity Checks e Nucleic acids extracted from cells are accompanied by protein and extensive purification is required to separate the protein impurity The 260 280 ratio gives an indication of purity it is only an indication however and not a definitive assessment Pure DNA and RNA preparations have expected ratios of gt 1 8 a
26. 446 4 0 542 4 wavelength 546 nm Absorbance 0 243 A Curve Fit Regression OOo coco Version 5 4 18 4 2011 Parameters Print Graph Sample Humber Save Method Anto Print 0 4 i p Ea ale F Fa fi E a n 04 DLE 08 10 Le 14 Biuret 4 Concentration pa ml Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Range 0 001 to 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes Press OK to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Results screen Step 14 Insert the reference and press the GE key This will be used for all subsequent samples until changed Step 15 Insert the sample and press The concentration of the sample is taken and displayed Repeat step 15 for all samples Press to return to the Protein Folder Press C Options d to display available Options which are described helaw Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Displays the calibration graph cursors give values for last measured sample 7 Sample numbe
27. 6 SD Card Accessory 80 3005 00 Spare printer paper 80 3004 07 Excel Electronic Pipette 0 5 10ul with adapter for use with Picodrop P3600L 10 Manual Gilson Pipetman Pipette 0 5 10ul with adapter for use with Picodrop F144802 Interchangeable 10mm Cuvette block with lenses fibres P200Cuvette PicoCal UV Vis 260nm Wavelength Standard fluid 750ul tube PCALO1 GLP MAINTENANCE After Sales Support Support agreements that help you to fulfil the demands of regulatory guidelines concerning GLP GMP are available gt Calibration certification using filters traceable to international standards s Certificated engineers and calibrated test equipment Approved to ISO 9001 standard Choice of agreement apart from break down coverage can include Preventative maintenance Certification When using calibration standard filters insert such that the flat surface is facing away from the spring end of the cell holder Observe all necessary precautions if dealing with hazardous samples or solvents Lamp Replacement The xenon lamp should not need replacement until after several years of use In the unlikely event that it does need replacing this should be undertaken by a service engineer from your supplier Cleaning and general care of the instrument External cleaning Switch off the instrument and disconnect the power cord Use a soft damp cloth Clean all external surfaces A mild liquid detergent may be used to remove stubborn marks Quic
28. A F E D4 0 6 08 Lo Le 1 4 Version 5 4 18 4 2011 Calibration Screen replicates off This shows the calibration values and allows standards to be measured Step 11 Insert the reference sample Press ED key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result Repeat step 12 for all standards A graph will display the results and the fitted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 When all standards are measured the OK box appears Press to accept the calibration an go to the Results screen see below OR Press Back to cancel selections and return to the Standards screen Calibration Screen replicates on This shows the calibration values and allows standards to be measured Step 11 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Press to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press to measure the standard and store the result Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arr
29. E Sample Number 8 Save method use the left and right arrows to select a folder E save Method to store in Favourites Methods 1 9 press the down arrow GP aute Print and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 40 Picodrop Ltd 2 BCA The procedure is as follows BCA Parameters Curve Fit Regression Calibration Replicates DP BCA Standards Version 5 4 18 4 201 Curve Fit Calibration Standards Replicatez 4 Step 1 Press 2 to select BCA mode Step 2 Wavelength for this stored method is pre set to 562nm Step 3 Enter the number of standard concentration points 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key lt gt and then use the left right arrows ug ml ug pl pmol ul mg dl mmol l umol g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen
30. Important if the fibres do not come away from the holder easily please seek assistance from Picodrop service team info picodrop com DO NOT PULL ON THE FIBRE OPTIC CABLES Check whether either lens at the end of each cable is wet This happens when excess sample is picked up on the outside of the tips Wash each lens with pure water and dry with tissue Use a cotton tip soaked in acetone to clean and dry the lenses Remove the round base from the holder by unscrewing Thoroughly rinse the metal base unit in pure water and then allow to air dry Re assemble unit and repeat tests as detailed above Lens Extraction tool Version 5 4 18 4 2011 Page 78 Picodrop Ltd SPECIFICATION AND WARRANTY Wavelength range Sample Containment Pathlength Recalibration Speed of sample measurement Nucleic Acid Detection Range cuvette Nucleic Acid Detection Range tip Protein Detection Range cuvette Protein Detection Range tip Monochromator Wavelength calibration Spectral bandwidth Wavelength accuracy Wavelength reproducibility Light sources Detector Photometric range Photometric linearity 220 950 nm Pipette Tip or 10mm Cuvette Not required as path length fixed by tip No moving parts 4 seconds 0 4 120 ng ul 3 1200 ng ul 0 02 2 5 mg ml 0 1 25 mg ml Flat grating Automatic upon switch on 5 nm 2 nm 1 nm Pulsed xenon lamp 1024 element CCD array 0 300 to 2 5004 0 to 199 T 0
31. Ltd 1 DNA The procedure is as follows Step 1 Press 1 to select DNA mode Step 2 Select path length using the left and right arrows Options are 1mm tip or 10 mm cuvette A Aarametsls Press the down arrow Step 3 dilution factor known Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace and clear the last digit entered Sie sm Step 3 calculate dilution factor r Press RED to enter the dilution factor screen Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow O o 2 Cancel Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press D to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Select the units of measurement using the left and right arrows Options ug ml ng ul ug pl Press the down arrow Step 6 Enter the factor using the keypad numbers Default value is 50 range is 0 01 to 9999 Step 7 Press OK D to enter the Results screen OR Cancel to return to the Nucleic Acids folder Results Screen Step 8 Insert the reference Press GE Key This will be used for all subsequent samples until changed Step 9 Insert sample and press Th
32. OK to store the chosen parameters or Cancel Step 5 To enter the results screen with the selected parameters press Cancel the selections and return to the Applications Folder by pressing Cancel Step 6 if using a Factor Insert the reference Press Blank key This will be used for all subsequent samples until changed STEP 7 Insert sample and press The concentration of the sample is displayed Results shown as indicate the concentration is out of range Repeat step 7 for all samples Press to return to the Applications folder Press lt gt to display available Options which are described below Page 16 Picodrop Ltd Concentration Step 6 if using standard mode Wavelength Insert the reference Press GE key This will be used for all 260 mm subsequent samples until changed Pr Press D to display the Run Standard screen ae Runihestandard by pressing ae OR Press cancel to return to the measure screen umol Concentration STEP 7 Insert the sample and press DJ The concentration of the sample is displayed Results shown as indicate the concentration is out of range wavelength 6 nm Z TT 300 Repeat step 7 for all samples Press to return to the Applications Folder Press C Options to display available Options which are described below Options select using key pad numbers Return to parameters screen step 1 above E Parameters 2 Print result via selected method P
33. Picodrop Ltd 5 Biuret The procedure is as follows Biuret Parameters Step 1 Press 5 to select Biuret method Curve Fit Step 2 546 nm Wavelength for this stored method is pre set to 546 nm Step 3 Standards Calibration Enter the number of standard concentration points 1 9 to be Standards used in the curve using the keypad numbers or left and right arrows Replicates Press the down arrow Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters or Cancel Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin Biuret Parameters interpolated or cubic spline Press the down arrow Curve Fit Step 6 546 nm Select the calibration mode either standards measure prepared standards or manual keypad data entry Standards Step 7 if standards selected Standards Select th
34. QPico20 USER MANUAL PO Box 188 Saffron Walden CB10 9BA United Kingdom Tel 44 0 131 2080522 info picodrop com www picodrop com All product and brand names used in the document are trademarks or registered trademarks of their respective holders Copyright 2011 Picodrop Ltd Registered in England no 05505772 Part no 41 56 1883 TABLE OF CONTENTS DECLARATION OF CONFORMITY oscar E saute 4 ESSENTIAL SAFETY ING Wa erate tae crc ds 5 Unpacking Positioning and Install lO uni nnmnnn nnna 5 INTRODUC TION zaira a a a a a io cid 6 OUF SPECTFODMOLOMCLOR Fae a aaa a a Eae Aaaa 6 Sample Nanding UDS a a a i 6 C rrectPipetino Procedure aria a a a a 7 Instrument S t OD zracima a 8 KeVOad and GIS play scne a Nea 10 SONWArE SIVIC cacian a a E a 12 THE APPLICATIONS FOLDER 0 ao 13 SUMMARY aiie E a E a E a 13 1 Single Wavelength Abs and T s sssnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnmnnn nnmnnn nnmnnn 14 2 CONGCCIATAUION vr aaa aaa aaa aaa aE it a A 16 De Wave SCI osora a aao la iaaa laa e aai A aaa 18 4 SIMPIC KNEG Su A A it 21 LAN AGUA A dt AE aSa 24 6 NIUITIDIC Wavelengths A A 28 T ADSORDANGCE RA Oia A di 29 TPE TIPE SCIENCE FOLDER ola a cous ucuagus saeeecumucaseeids 31 A eee ee ee eee ee eres eee 32 PARNA erent ene ae ee aE A eee 34 A O eee TL ee et et ee cena 36 Protein DET MINA ON ai 38 o A e 39 2 BO A eg a a ee ee ee ee ee eee eee 41 A ONG ai sates vatican acct g
35. ameter screen Standard Curve Calibration Calibration Screen replicates off This shows the calibration values and allows standards to be o wo IESS measured Step 10 Insert the reference Press Blanki key This will be used for all subsequent samples until changed Step 11 Insert the standard use C to clear previously stored results before measuring 1d 20 30 340 5O E Press D to measure the standard and store the result Repeat for all standards Standard Curve Calibration Standards e A graph will display the results and the fitted curve as the 10 0 0 0154 Pa measurements are input 70 0 0 049 A 31 0 0 0824 El Use the up and down arrows to select a standard to be repeated 62 0 mts E if a poor reading has been obtained Use C to clear the previous 0 04 E reading o 02 E a Step 12 nog 10 20 30 ug 50 El Press OK to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Version 5 4 18 4 2011 Page 25 Picodrop Ltd Version 5 4 18 4 2011 Calibration Screen replicates on This shows the calibration values and allows standards to be measured Step 10 Insert the reference Press cE key This will be used for all subsequent samples until changed Step 11 Press to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press Enter to measure the standard an
36. ancel selections and return to the Standards screen 0 20 0 40 0 60 0 60 1 00 1 40 Calibration Screen replicates on This shows the calibration values and allows standards to be measured Step 11 Insert the reference sample Press EE key This will be used for all subsequent samples until changed Step 12 is 04 DE 08 40 42 14 Press to display the replicate entry boxes Use C to clear previously stored results before measuring Lowry Calibration Insert the standard and press to measure the standard and store the result Replicates 3 a Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading 2 0 4 0 6 08 40 Le L4 Step 13 Press to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Version 5 4 18 4 2011 Page 48 Picodrop Ltd Calibration Manual entry Shows previously entered calibration values and allows values to o be entered via the keypad 0 017 A Y The highlighted box can be edited in order to enter an 0 056 4 _ ae fl absorbance value corresponding to a given concentration value 0 094 4 eS using the keypad numbers Range 0 001 to 9999 Use C to 0432A gin fimp backspace and clear the last digit entered and the up a
37. as much smaller optics than most conventional spectrophotometers and more light is transmitted through to the detector resulting in lower than expected OD 600 values Results obtained by comparing measured OD 600 with expected OD 600 see above indicate that a correction factor of 2 0 is required to make the data comparable to larger instruments this factor is included as a default value in set up The use of 10 mm pathlength disposable cells is recommended for optical density measurements of cell culture solutions to prevent the suspension settling too quickly and giving an OD that changes with time glycerol should be added to the sample The procedure is as follows OD 600 Parameters Step 1 Wavelength Select the wavelength Default value is 600 nm Press the down arrow Step 2 Correction Enter the factor to compensate for different optical configurations 2 00 between this and other instruments Default value is 2 Press the down arrow Step 3 Select the units Options are OD or cells ml If cells ml is selected two further parameters are displayed OD 600 Parameters Step 4 if cells ml selected Enter the factor using the keypad numbers Range 0 00 to 9999 VSEE Factor C button backspaces and clears the last digit entered Press the down arrow Step 5 if cells ml selected Correction Multiplier Select the multiplier using the left and right arrows Options are 1000 or 1 000 000 Step 6 Units D Press OK to e
38. asurement if allowed to rise more than 2mm up the tip It is preferable that the sample tip and pipette holder are allowed to equilibrate to room temperature for 5 minutes before commencing measurement The detection point of the tip is 1 5mm from the end of the tip so it is best procedure to try not to submerge the tip more than 1mm into your solutions otherwise tip wiping may be necessary It is optional whether you decide to use the same tip for both blank and sample or a different tip each time The special ultra low retention tips will ensure zero carryover of blank to your sample and so using the same tip is a safe operation Using different tips for blank and sample will also provide results which meet the published specification but does however introduce an additional variable which may become significant with very low concentration samples e lt 10ul ng DNA l Recommended pipetting procedure Version 5 4 18 4 2011 Page 7 Picodrop Ltd Instrument Set Up Hardware installation If not already pre assembled The pipette holder has been dismantled and disconnected for shipment Fix the pipette holder to its round base by simply screwing it on Connect the cables to the base of the pipette holder Push the connector into the socket and hand tighten the nut to hold the connector in place Do not use tools to tighten the fixing nut Connect the base plate to the underside of the unit with the 2 hex head screws provided V
39. avourites NOTE any methods currently on the instrument will be overwritten by the restore process and folder names will be changed to match those being restored Transfer of data to PC and file management Methods data and method folders stored on the SD card can be archived to PC the data is accessed on the PC using the Print Via Computer PVC software supplied with the instrument and it can then be printed or saved in a variety of formats graphics text or Excel Refer to the user manual included on the PVC CD for installation and operating instructions of PVC To transfer data to a PC the SD memory card should be inserted into the relevant SD memory card slot either on the PC or on an SD card reader connected to the PC PVC Viewer is started and data opened using the Load File menu navigating to the 1Serial no PVC directory on the SD memory card and selecting the relevant file Alternatively you can double click on the file Individual method files can be manipulated on the PC using Windows Explorer Files can be deleted renamed maximum 24 characters or moved between folders maximum 9 methods per folder NOTE Folder names and method names edited on the PC are not restored to the instrument Folders on the instrument should be renamed using options Folder Names Restoring method folders to multiple instruments cloning Using this function allows all instruments to be set with the same folder and method structure in a m
40. be stored in the folder of eG ea Methods 1 Methods 2 Methods 3 of 9G ea Methods 4 Methods E Methods E of eG ob hMethods Y Methods E hMethods 3 Methods These are further storage folders enclosed in the top level Methods folder Up to 9 Methods may be stored in each folder Operation is identical to the Favourites Folder Saved methods can be locked unlocked and deleted using the Options menu Select the method by pressing the relevant key pad number and then press the lt gt key Delete Method Press 1 to select delete method Select the method to be deleted using the left and right arrows Press D to delete the method Methods Methods 1 OR amp cancel to return to Favourites Methods folder 1 Lock Method O A Press 2 to select lock method Single wavelength Select the method to be locked using the left and right arrows Press the down arrow Select a pass code using the keypad numbers or left and right arrows Press O to lock the method 1 Delete Method E Lok Method OR S cancel to return to the Favourites Methods folder Unlock Method Unlock Method Press 3 to select unlock method select the method to be unlocked using the left and right arrows Press the down arrow Enter the pass code using the keypad numbers or left and right arrows Press O to unlock the method OR cancel to return to the Favourites Methods folder Version 5 4 18 4 2011 Page 55 Picodrop Ltd UTILITIES
41. bers or left and right arrows Replicates Press the down arrow Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol l g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters or Cancel Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin Curve Fit interpolated or cubic spline Press the down arrow Lowry Parameters Step 6 Standards Calibration Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates using the left and right arrows ee This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 nem 2 oF 3 Step 8 Press Next to enter the Standards screen OR Press Cancel to cancel selections and return to the Protein folder Standards Screen
42. bsorption value for a particular protein must be determined The presence of nucleic acid in the protein solution can have a significant effect due to strong nucleotide absorbance at 280 nm This can be compensated by measuring Abs 260 and applying the equation of Christian and Warburg for the protein crystalline yeast enolase Biochemische Zeitung 310 384 1941 Protein mg ml 1 55 Abs 280 0 76 Abs 260 or Protein conc Factor 1 Abs 280 Factor 2 Abs 260 This equation can be applied to other proteins if the corresponding factors are known The instrument can determine protein concentration at 280 nm and uses the above equation as default the factors can be changed and the use of background correction at 320 nm is optional To customise the equation for a particular protein the absorbance values at 260 and 280 nm should be determined at known protein concentrations to generate simple simultaneous equations solving these provides the two coefficients In cases where Factor 2 is found to be negative it should be set to zero since it means there is no contribution to the protein concentration due to absorbance at 260 nm Set Factor 2 0 00 for direct A280 UV protein measurement Factor 1 is based on the extinction coefficient of the protein If BSA bovine serum albumin is an acceptable standard setting Factor 1 1 115 will give linear results from 0 to 0 8 mg ml protein Protein mg ml 1 115 Abs 280 Rapid measurements su
43. by measuring a standard of known concentration The procedure is as follows Concentration Parameters wavelength Units Mode Factor lt P DK Concentration Parameters Concentration Version 5 4 18 4 2011 Step 1 Set wavelength by using keypad numbers or left and right arrows Press the down arrow key Step 2 Select the mode Factor user entered or Standard factor is calculated from a calibration sample using the left and right arrows Press the down arrow key Step 3 if Factor is selected Enter the Factor using the keypad numbers Range 0 001 to 9999 Use the C button to delete the last digit entered Press the down arrow key Step 3 if Standard is selected Enter the concentration using keypad numbers Range 0 01 9999 Use the C button to delete the last digit entered Press the down arrow key Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press
44. ch as this at Abs 280 are particularly useful after isolation of proteins and peptides from mixtures using spin and HiTrap columns by centrifuge and gravity respectively Protein Determination at 595 546 562 and 750 nm The Bradford method depends on quantitating the binding of a dye Coomassie Brilliant Blue to an unknown protein and comparing this binding to that of different known concentrations of a standard protein at 595 nm this is usually BSA bovine serum albumin The Biuret method depends on reaction between Cupric ions and peptide bonds in an alkali solution resulting in the formation of a complex absorbing at 546 nm The BCA method also depends on reaction between cupric ions and peptide bonds but in addition combines this reaction with the detection of cuprous ions using bicinchoninic acid BCA giving an absorbance maximum at 562 nm The BCA process is less sensitive to the presence of detergents used to break down cell walls The Lowry method depends on quantifying the colour obtained from the reaction of Folin Ciocalteu phenol reagent with the tylsryl residues of an unknown protein and comparing with those derived from a standard curve of a standard protein at 750 nm this is usually BSA bovine serum albumin Detailed protocols are supplied with these assay kits and must be closely followed to ensure accurate results are obtained The use of plastic disposable cells is recommended To use a zero concentration standard include it in the n
45. d store the result Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 12 Press to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Range 0 001 to 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes Press OK to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Results screen Step 13 Insert the reference and press the GE key This will be used for all subsequent samples until changed Step 14 Insert the sample and press DJ The concentration of the sample is taken and displayed Repeat step 14 for all samples Press to return to the Applications Folder Press C Options d to display available Options which are described below Page 26 Picodrop Ltd Standard Curve Calibration Standard Curve Calibratio Version 5 4 18 4 2011 oo es ss ss Pi
46. d y axes Zoom mode This sets up the operation of the Zoom keys up and down arrows x amp y axes expands the display around the cursor measurement point whilst the other options select the absorbance or wavelength axes respectively With x or y axis limits set to on Zooming out will only be permitted to the set limits x y axis limits Setting x or y axis limits to On activates the start and finish points of the desired graph to user defined specific wavelengths and or absorbance values Pressing Cancel ignores the selection pressing accepts them and displays the required graph Picodrop Ltd 4 Simple Kinetics Kinetics studies where the change in absorbance needs to be followed as a function of time at a fixed wavelength can be readily performed Reagent test kits are routinely used for the enzymatic determination of compounds in food beverage and clinical laboratories by measuring NAD NADH conversion at 340 nm The change in absorbance over a specified time period can be used to provide useful information when an appropriate factor defined in the reagent kit protocol is applied Reaction rate and enzyme activity can be calculated if the factor used takes account of the absorbance difference per unit time as opposed to the absorbance difference per se For this reason the change in absorbance per minute AA min concentration AA min x factor and correlation coefficient calculated from a bes
47. do a reference scan Enter Enter or confirm a selection Take a measurement Keypad and display The back lit liquid crystal display is very easy to navigate around using the alphanumeric entry and navigation arrow keys on the hard wearing spill proof membrane keypad On off Arrow keys O bG Escape Cancel A A Set reference View Enter selection take measurement Version 5 4 18 4 2011 Page 10 Picodrop Ltd Options select using key pad numbers E Parameters 1 View parameters for the experiments O Prin 2 Print the results Print Dats 3 4 5 6 Described in the application P seco At Cursor 7 Define the sample number you wish to start from GP Set tn At Cursor 8 save the parameters as a method to a defined folder C Slope name with a defined method name E Sample Number 9 Toggle auto print on off Default is off E Save Method i E to Prin a Exit options by pressing or wait Experienced operators can use the numeric keys as a shortcut to the option reauired without needina to enter the Ontions menu Version 5 4 18 4 2011 Page 11 Picodrop Ltd Software style The user interface is built around having folders of files which are displayed on the home page when the instrument is switched on Different folders are numbered and opened by using the associated number key on the keypad ol Applications Favourites Methods oQ el Utilities Life Science 41107
48. e contacts facing towards the user and the cut out corner on the right hand side i e downwards When a compatible SD card is inserted into the accessory the red light flashes momentarily and the SD memory card icon appears on the instrument home page Life Science Applications oQ eQ oQ Methods Utilities SO Memon Card Saving methods to SD memory card When an SD memory card is inserted into the accessory it is possible to save methods directly to the card Methods are stored on the card in a directory named Instrument Type Methods where instrument type will be dependant upon instrument being used this directory structure is evident when the SD card is connected to a PC To save a method to the SD memory card the instructions for the relevant application from the instrument user manual must be followed Typically press the Options button or relevant numerical short cut press Save Method use the right and left arrows to select the folder on the SD memory card to which you wish to save the method change the filename if required press Save Single Wavelength 2 5 Save Method 1 0 Folder q Methods 4 d Method Name Single wavelength Save ER Cancel NOTE a maximum of 9 methods can be stored in the SD memory card folder and in the nstrument Type Methods directory These stored methods can also be opened on different instruments and then stored into other method folders if required Version 5 4 18 4 2011
49. e number of replicates using the left and right arrows This determines the number of standards to be measured and Units Replicates averaged at each standard concentration point Can be OFF 1 wat rn 2 0r 3 Step 8 Press Next to enter the Standards screen OR Press Cancel to cancel selections and return to the Protein Biuret Standards folder Std 1 Std 4 a Standards Screen Std 2 Std 5 Step 9 aoe i Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears Std 3 Std 6 0 60 madi the last digit entered Step 10 Press Next to enter the Calibration screen OR Press Back to return to the Parameter screen Version 5 4 18 4 2011 Page 50 Picodrop Ltd Calibration Screen replicates off This shows the calibration values and allows standards to be measured Biuret Calibration Step 11 o Insert the reference Press GEM key ooa 3 This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result Repeat step 12 for all standards A graph will display the results and the fitted curve as the measurements are made i a 04 DE DE Lo le 1 4 Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C
50. ersion 5 4 18 4 2011 Page 8 Picodrop Ltd Place the holder on the base plate and connect fibre cables to the unit connectors Slide the provided PIPETTE GUIDE collar onto the pipette The easiest way to attach the collar is to place the collar in the tall pipette holder and then insert the pipette until the collar has located on the pipette Remove the pipette and the collar will remain attached to the pipette The collar can be easily removed for cleaning if necessary Version 5 4 18 4 2011 Page 9 Picodrop Ltd Key Action On off key Turns the instrument on off Arrow keys Use the four arrow keys to navigate around the display and select the required setting from the active highlighted option View Options lt gt View options for that application mode Some of these are common to all applications and described below Options unique to an application are described in the relevant section Alphanumeric keys Use these to enter parameters and to write text descriptions where appropriate or required Use repeated key presses to cycle through lower case number and upper case Leave for 1 second before entering next character Use C button to backspace and 1 to enter a space Escape Cancel Escape from a selection and return to the previous folder Stop making measurements Set Reference GE Set reference to 0 000 A or 100 T on a reference solution at the current wavelength in the mode selected When in scan mode
51. g methods from SD Memory Card viii andes eed a i 72 Saving datato SD memory cada iaa 72 Backup Ol method tolders ica ala 74 Restoring Method TOlderS cirio aaa 75 Transfer of data to PC and file ManageMent ooooncnncccccnccnnccccnnncnnnnnnnnnconnnonnnncnnnnnnnnnrnnnnnnnanrnnnnnnanarannns 75 Restoring method folders to multiple instruments ClONINQ ooocccnccccconcnnnonnnancnnncnnnnncnnnnonnnnennnnnnnos 75 PRINT VIA COMPUTER iccicocc ios As ae ae ee ores Ud 76 ACGESSORIES ette tdo nee nee ici 77 MAINTENANCE cuco asin a o o e e do eee ae 77 Aner Sales SUD WON aii cdi 77 tamp ROCA ia 77 Cleaning and general care of the instrument cccceeeeseeeeeeeenneeeeeeeenseeeseeeesseeeeeoeesseeeeeoensseeeseoensseeesees 77 SEFVice Glean PLrOCCQUEC susine ded iaaa i aa Aaaa aA aaa Aa A aaa aaa aS 78 SPECIFICATION AND WARRANTY ici tii 79 WARRAN sc li la 80 Fibre Optic Cable Handing and Limits of Warranty ooooconnnccnncccnnncnnnnccnoccnonncnnnnncnnnnnonnnnnnnncnnrnnnnnnnnnnnennnnnnas 80 Version 5 4 18 4 2011 Page 3 Picodrop Ltd DECLARATION OF CONFORMITY This is to certify that the following products conform to the requirements of the following Directives Pico200 Micro volume spectrophotometer part numbers P200 P200P P200BT P200SD P200E P200PE P200BTE P200SDE 2006 95 EC Low voltage equipment safety directive 2004 108 EC EMC directive 2002 96 EC amp 2003 108 EC EC Directive on Waste Electrical and Electronic Equipment
52. ges are discovered inform your supplier immediately e Inspect the instrument for any signs of damage caused in transit If any damage is discovered inform your supplier immediately e Ensure your proposed installation site conforms to the environmental conditions for safe operation e Indoor use only e Temperature range 5 C to 35 C Note that if you use the instrument in a room subjected to extremes of temperature change during the day it may be necessary to recalibrate by switching off and then on again once thermal equilibrium has been established 2 3 hours e Maximum relative humidity of 80 up to 31 C decreasing linearly to 50 at 40 C e The instrument must be placed on a stable level bench or table that can take its weight lt 4 5 kg so that air can circulate freely around the instrument e This equipment must be connected to the power supply with the power supply unit and cord supplied It can be used on 90 240 V 50 60 Hz supplies e Ifthe instrument has just been unpacked or has been stored in a cold environment it should be allowed to come to thermal equilibrium for 2 3 hours in the laboratory before switching on This will prevent calibration failure as a result of internal condensation e Switch on the instrument via the keypad after it has been plugged in The instrument will perform a series of self diagnostic checks e In accordance with good laboratory practice it is recommended that periodic performance
53. he left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 23 Picodrop Ltd 5 Standard Curve The construction of a multi point calibration curve from standards of known concentration to quantify unknown samples is a fundamental use of a spectrophotometer this instrument has the advantage of being able to store this curve as a method using up to 9 standards To include a zero concentration standard include this in the number of standards to be entered and enter 0 00 for concentration use a reagent blank when required to enter the zero standard The procedure is as follows Standard Curve Parameters wavelength Curve Fit Step 1 i Select the wavelength using the keypad numbers or left and right arrows EEFE Press the down arrow eee Enter the number of standard concentration points to be used in the curve 1 9 Press the down arrow Replicates Step 3 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note
54. he up and down arrows to move between boxes Press OK to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Results screen Step 14 Insert the reference and press the GE key This will be used for all subsequent samples until changed Step 15 Insert the sample and press DJ The concentration of the sample is taken and displayed Repeat step 15 for all samples Press to return to the Protein Folder Press C Options d to display available Options which are described helaw Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Displays the calibration graph cursors give values for last measured sample 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 46 Picodrop Ltd 4 Lowry The procedure is as follows Lowry Parameters Step 1 Press 4 to select Lowry method Curve Fit Step 2 250 nm Wavelength for this stored method is pre set to 750 nm Step 3 Standards asa Enter the number of standard concentration points 1 9 to be Standards used in the curve using the keypad num
55. ia a io m m o so Er Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Displays calibration graph cursors give values for last measured sample 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 4 Print graph using selected method Grayed out if no data are available 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 27 Picodrop Ltd 6 Multiple Wavelength This makes up to 5 absorbance measurements on the same sample The procedure is as follows Multi wavelength Parameters Step 1 Select the number of wavelengths Wavelengths a3 ki Step 2 Enter the first wavelength using either the number keys or the Press the down arrow
56. in RNA purification and which absorbs over the 230 260 nm range A wavelength scan of the nucleic acid is particularly useful for RNA samples e The instrument can display 260 280 and 260 230 ratios and compensates for dilution and use of cells that do not have 10 mm pathlength dilution factor and cell pathlength can be entered Use of Background Correction e Background correction at a wavelength totally separate from the nucleic acid and protein peaks at 260 and 280 nm respectively is sometimes used to compensate for the effects of background absorbance The wavelength used is 320 nm and it can allow for the effects of turbidity high absorbance buffer solution and the use of reduced aperture cells The instrument can use background correction e _ If it is used there will be different results from those when unused because Abs320 is subtracted from Abs260 and Abs280 prior to use in equations Concentration Abs 260 Abs 320 Factor Abs ratio Abs 260 Abs 320 Abs 280 Abs 320 Abs ratio Abs 260 Abs 320 Abs 230 Abs 320 e f your laboratory has not used background correction before set this option to NO e The use of background correction can remove variability due to handling effects of low volume disposable cells Note e absorbance maximum near 260 nm and absorbance minimum near 230 nm e flat peak near 260 nm and steep slope at 280 nm e very little absorbance at 320 nm Version 5 4 18 4 2011 Page 31 Picodrop
57. is measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 absorbencies are calculated both corrected by the background wavelength value if selected Gives concentration based on absorbance at wavelength 1 Repeat step 9 for all samples Press to return to the Nucleic acid folder Press lt gt to display available Options which are described below Version 5 4 18 4 2011 Page 32 Picodrop Ltd Options select using key pad numbers 1 Return to parameters screen step 1 above dh Parameters 2 Print result via selected method O Pin 3 Toggle graph on off The graph shows a wavescan plot Graph across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if background correction selected 320 nm 7 Sample number add a prefix to the sample number and O Sample Number reset the incrementing number to the desired value O Save Method 8 Save method use the left and right arrows to select a folder a autori 7 to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 33 Picodrop Ltd 2 RNA The procedure is as follows Step 1 Press 2 to select RNA mode Step 2 Select path length using the left and right arrows Options are 1mm tip or 10 mm cuvette Press the down arrow Pathlength Units Step 3 dilution fact
58. k Clean procedure for sample holder In the event that sample leaks from a pipette tip or dust reduces the light transfer through the pipette holder simply unscrew the silver cable fibres from each side of the silver pipette holder no tools required silver screws should be only hand tight Unscrew the circular base from the tube section Unscrew the single screw on the tube to release the main tube from the bottom tip holder Either soak the holder in hot water with detergent for 30mins and air or drip dry or alternatively simply wash with an ethanol or similar solvent Reassemble and re test instrument If this quick clean procedure does not improve the results please follow the Service Clean procedure described in the following section Important Please note that as the lenses get dirty with use then the stability will decrease and this will accentuate any background noise and small variations between tips Version 5 4 18 4 2011 Page 77 Picodrop Ltd Service Clean Procedure Detach the Silver fibres by unscrewing from each side of the silver pipette holder Use the 1 5mm Allen key as supplied to loosen the 2 fibres by turning the two sunken screws in the front of the bottom of part of the pipette holder and then unscrew the two fibres away from the holder and attach the LENS EXTRACTION TOOL pictured below as supplied to each lens in place of the fibre and pull firmly on the lens extraction tool to pull out lenses
59. k saa Se atin de sade a oe ae cee kee engl es cee ge alae 44 AEN SOWIE add dia daa 47 A eee eee eee nea ee e ee eee ee 50 Bacterial Cell Culture Measurement ODG600 cccceceeeseeeesecennseeeeeeenseeeeseoessseeeseoeaseeeesooanseeessoonneees 53 FAVOURITES AND METHODS FOLDERS scsiicianiina as 55 FAVOR ia 55 An nn E E E 55 UTILITIES FOLDER mcin ni 56 AA O 57 Date and TIME arsena ias 57 ZAC GLO i ell ae a a cin het bana ceantsina Sedu mecneveunnna tiara nates encase 57 A e e eee eee 58 real a 22 A A E A ee ee mee ee ee ee ene 59 A een cae eee cae heen tate ae ce Seatac mmrae eae aaa aaa 59 FO NAMES cias 59 A Pea Ee Se Emm ere Nn ee a ee ene eee eee ee 60 GANIC Siete stp ae estate o eal Nl od do Salut e 60 ACCESSORIES INSTALLATION iii a aa aaa sud a a dai venwdesaste lain E ids 62 Printer ASANO A A 62 Loading changing the printer paper cid 64 Changing to the Bluetooth Accessory sssssensnunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnmnnn nnmnnn nnna 65 SD MEMORY CARD ACCESSORY sorsas es a Ea aa aea aa ae S o a aG 68 AU ARPA aaa a aaa aaraa E a aaa aa eTa aS aE 68 A a A a a ech huis attic ala Ue 68 SA OLY sierra tie sat ie tual ae Rn A eS a lien eit bea cee ot alia oan ec ial ce aie 68 FITUAG SD memory card ACCESSO iini een ieee alee ee 69 Version 5 4 18 4 2011 Page 2 Picodrop Ltd OPERATION iii A antic da sais A A A a vandaag laces 71 saving methods to sD iMeMOry Cal io 71 Loadin
60. kground correction is On Enter the third wavelength from which the background correction will be obtained Step 5 Press Next O to enter the Parameters screen OR Press Cancel to return to the Applications Folder Absorbance Ratio Parameters Absorbance Ratio Parameters Screen Pathlength Factor Step 6 Select the pathlength 5 or 10 mm using the left and right arrows Dilution Factor Press the down arrow 1 00 Step 7 Dilution Factor known Enter a dilution factor by using the keypad numbers within the range 1 00 9999 OR Step 7 Calculate Dilution Factor Press the options key Enter the volume of the sample range 0 01 9999 using the keypad numbers Press the down arrow Enter the volume of diluent range 0 01 9999 by using the keypad numbers Dilution Factor Press OK O to calculate the dilution factor and return to the Parameters screen or press Back to cancel selections Step 8 Select units of measurement using left and right arrows Options are ug ml ng ul ug pl Press the down arrow Yolume Diluent Step 9 Enter the factor using the keypad numbers Range 0 001 to 9999 Press OK O to enter the results screen or Cancel Y to return to the Applications Folder gt Cancel gt o F Version 5 4 18 4 2011 Page 29 Picodrop Ltd Results Screen Step 10 Insert the reference Press GE key This will be used for all subsequent samples until changed Step 11
61. l to return to the Utilities folder without storing the settings Version 5 4 18 4 2011 Page 57 Picodrop Ltd 3 Printer Sets up printing options Select whether auto print is on or off using the left and right Printer arrows When auto print is on the results are automatically printed after a measurement is taken When it is off printing has to be initiated manually This can also be set using the Options Auto Print key GED in cach application or method The default is OFF Press the down arrow Select how the data are sent Options are Built in internal printer or to a computer via USB port or Bluetooth Printer Press OK to store the settings and return to the Utilities OR OR Press Cancel to return to the Utilities folder without storing the settings Cancel The procedure is as follows Version 5 4 18 4 2011 Page 58 Picodrop Ltd 4 Preferences Sets user preferences The procedure is as follows 5 Contrast Preferences Auto Standby o History Select games function This determines whether the games folder is displayed or not Options are yes or no Press the down arrow Define the screen layout of folders Options are either a grid format default or a list Press the down arrow Select whether to use previously entered parameters on switch on or use defaults Press the down arrow Select whether to use a standby mode after defined periods Options are 1 hour 2 hours at night or
62. n be used is approx 10ul in a Quartz ultra micro cell e 12mm test tubes may be used e g for cell cultures however they are not recommended as higher quality data is produced by using disposable cuvettes for the analysis If used align the indicator line on 12 mm test tubes in the same direction to ensure reproducible positioning of the tube Note that test tubes do not last forever and that the surface becomes scratched and blemished through repetitive use if this is the case they should be replaced Version 5 4 18 4 2011 Page 6 Picodrop Ltd Correct Pipetting Procedure Vortex sample briefly 5 20secs Spin down samples briefly 10 15secs in a microfuge Using a P10 pipette and a UVpette tip pipette your sample A minimum volume of 2 5ul is recommended To minimise solution on the outside of the tip avoid submerging the tip too far below the sample meniscus If necessary Wipe off excess liquid from outside of tip with a dry lint free tissue this is particularly important if using viscous protein solutions Be careful not to touch the bottom of the tip as the sample may be drawn out by the tissue Do not place UVpette tips or your sample too close to heaters or the fan of the PC as heating the tips or sample may result in a rapid contraction in volume once the tip is placed the cooler pipette holder This sample contraction will result in a space or bubble being visible at the bottom of the tip This space may interfere with sample me
63. nd gt 2 0 respectively deviations from this indicate the presence of impurity in the sample but care must be taken in interpretation of results e The 260 nm reading is taken near the top of a broad peak in the absorbance spectrum for nucleic acids whereas the 280 nm reading is taken on a steep slope i e small changes in wavelength cause large changes in absorbance Consequently small variations in wavelength at 280 nm will have a greater effect on the 260 280 ratio than variations will at 260 nm Thus different instruments of the same and different types may give slightly different ratios due to variations in wavelength accuracy But each instrument will give consistent results within itself e Concentration also affects 260 280 readings If a solution is too dilute the readings will be at the instrument s detection limit and results may vary as there is less distinction of the 260 peak and 280 slope from the background absorbance This is one reason why the Abs260 value should be greater than 0 1 for accurate measurements e An elevated absorbance at 230 nm can indicate the presence of impurities as well 230 nm is near the absorbance maximum of peptide bonds and also indicates buffer contamination since This EDTA and other buffer salts absorb at this wavelength When measuring RNA samples the 260 230 ratio should be gt 2 0 a ratio lower than this is generally indicative of contamination with guanidinium thiocyanate a reagent commonly used
64. nd down 0 369 A f arrows to move between boxes e 0 205 A F ri Press OK O to accept the calibration and go to the Results se 0 4 0 6 D B co La Ll4 screen see below OR Press Back to return to the Standards screen Lowry Calibration Standards 4 4 E mA es a a E 0 2054 0 05 Results screen Step 14 Insert the reference and press the GE key This will be used for all subsequent samples until changed Absorbance Step 15 0 095 A wavelength T50 nm Insert the sample and press The concentration of the sample is taken and displayed Repeat step 15 for all samples Curve Fit Interpolation Press to return to the Protein Folder Press lt gt to display available Options which are described helaw Options select using key pad numbers 1 Return to parameters screen step 1 above E Parameters 2 Print result via selected method Pin 3 Toggle graph on off Displays the calibration graph cursors EY Graph give values for last measured sample 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder EY Sample Number to store in Favourites Methods 1 9 press the down arrow Save Method and enter name O Auto Print 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 49
65. nt 6 Displays Graph Scale Parameter Screen See description below 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Peak Detection Shortcut button 4 Peak Detection AutoDetect Peaks Turns on and off the automatic peak AutoDetect Peaks Peak Detect on Zoom detection The following options determine how peaks are detected Minimum peak height Minimum height the peak has to be TE ieee A BON naan above the higher of the two adjacent minima for the peak to be detected Minimum peak width Minimum width of the peak as determined by the difference in wavelength between the higher of the two adjacent minima and the opposing intersection of that AA higher minimum level and the peak profile See the screen gt ox S cance displayed below Peak Detect on Zoom Determines whether peaks are re 0 5 assessed and tabulated when the user zooms into a region of the wavescan If off leaves the peak detection as determined on the un zoomed display 0 4 l i 0 3 ll Sort peaks by Determines the sequence that peaks are reported by Can be wavelength peak height or peak width og 440 p 480 o Draw Peaks Switches display of peak cu
66. nter the Results screen OR Press Cancel to cancel selections and return to the Life Science folder Version 5 4 18 4 2011 Page 53 Picodrop Ltd Results Screen Step 8 wavelength Insert the reference and press the 0A 100 T key This will be 600 nm used for all subsequent samples until changed Step 9 Absorbance Insert the sample and press DJ a The wavelength absorbance and OD600 value is displayed Repeat step 9 for all samples Press to return to the Life Science Folder Press lt gt to display available Options which are described below Options select using key pad numbers oO dels Return to parameters screen step 1 above Pin 2 Print result via selected method 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow E Sample Mumber and enter name EY Save Method 9 Auto print toggles auto print on off E Awuto Print y Exit options by pressing or wait Version 5 4 18 4 2011 Page 54 Picodrop Ltd FAVOURITES AND METHODS FOLDERS These folders are the storage locations for any user modified Applications Methods that are saved in the Options menu Both are accessible from the home folders page Favourites This folder enables the user to quickly select any frequently used Methods Up to 9 Methods may
67. ntly used configured methods nine methods per folder Utilities Instrument set up date time language etc and games Life Science Standard Life Science methods such as nucleic acid assays protein assays and cell counting The instrument is supplied with a program PVC Print via Computer on the accompanying CD When used with a USB cable to connect to a PC onto which the software has been installed it enables the user to print through the PC directly to the printer that is connected to it The data may also be stored as an Excel spreadsheet as an EMF graphics file a comma delimited csv data file a tab delimited txt data file or in native PVC format for later access Alternatively results may be sent to the PC via a Bluetooth accessory this can either be supplied pre installed or is available as an optional accessory if the need for its use arises after installation of the product PVC works in a similar way A printer is available for the instrument this may either be supplied pre installed or is available as an optional accessory if the need for its use arises after installation of the product Sample handling tips e Note that the light beam is directed from RIGHT to LEFT through the tip or cell chamber therefore please ensure the cell is inserted in the correct alignment e The optional cell holder accepts standard 10 mm pathlength quartz glass or plastic cells e The optical height is 15 mm and the minimum volume that ca
68. off Press OK to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the settings Ambient temperature can affect the display This function can optimise the display for local conditions The procedure is as follows Contrast Contrast Brightness 6 Folder Names Adjust the contrast using the left and right arrows Press the down arrow Adjust the brightness using the left and right arrows Press the down arrow Press OK to store the settings and return to the Utilities folder This folder allows you to rename the method or favourite folders Folder Hames Folder Hew Hame Te OF SS Cancel Version 5 4 18 4 2011 Select the folder you wish to rename using the left and right arrows Press the down arrow Input the new name for the folder Press OK to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the settings Page 59 Picodrop Ltd 7 About O Serial Number 54321 Version 4110 7 1 Displays the instrument serial number and software version Press OK O to close the window and return to the Utilities folder 8 Games om eff Spectra Blacks Sudoku 1 Spectroblocks 1 Hew Game 2 Options 3 Exit Classic block dropping game Follow the instructions Press Cancel to return to the Utilities folder without storing the set
69. omputer Bluetooth Version 5 4 18 4 2011 Page 67 Picodrop Ltd SD MEMORY CARD ACCESSORY The SD memory card accessory can be fitted to a Pico200 spectrophotometer for the following operations To export data from the instrument for loading into a PC To save methods To create a backup of all instrument methods for future restore To backup a specific instrument and restore these methods to other instruments in order to allow cloning The SD memory card accessory is compatible with the following types of storage media e SD memory cards e SDHC memory cards Data is stored on the SD memory card in a proprietary format with a pvc file extension and this data can only be accessed the Print Via Computer PVC software supplied with the accessory This software should be installed onto a PC as detailed in the PVC user manual stored on the PVC CD Installation Unpacking e Remove the accessory from its packaging and inspect it for signs of damage e Within the SD memory card accessory you will have the following e SD memory card PCB Module e New accessory cover e SD memory card e PVC Print Via Computer software CD e Allen key e lfthere are any signs of damage to the accessory of if any of the above components are missing please contact your supplier immediately Safety Read the safety instructions in the relevant instrument user manual Disconnect all power from the instrument before fitting the SD memory card accessory
70. or Nucleic Acids the LED next to the SD memory card will stay on continuously until the complete set of results is finished To close the results file the application should be exited using the ESC key in the usual way Removing the card whilst the LED is on will corrupt the collected data set All results are stored as an individual file in a directory called Serial no PVC on the SD memory card this directory structure is evident when the SD card is connected to a PC Results are identified by the format of the filename application type followed by an incrementing file ID For example DNA A001 PVC for a DNA file BCA001 PVC for a BCA Protein file For applications that print whole documents in one go such as Wavescan or Kinetics the LED next to the SD memory card will go off after saving each measurement In this case the card can be removed when the light is off without leaving the application Each result is stored as an individual file in a directory called Serial no PVC on the SD memory Card this directory structure is evident when the SD card is connected to a PC Results are identified by the format of the filename application type followed by an incrementing file ID For example WAVE 001 PVC for a Wavelength scan KINET001 PVC for a Kinetics file Version 5 4 18 4 2011 Page 73 Picodrop Ltd Backup of method folders This function allows the user to make a copy of installed methods on an instrument these methods can be restored
71. or known Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace and clear the last digit Dilution Factor Factor entered OR Step 3 calculate dilution factor Press EXP to enter the dilution factor screen a Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 gt OK Press the down arrow Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 RHA Parameters Press to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Select the units of measurement using the left and right arrows Options ug ml ng ul ug pl Press the down arrow Step 6 Enter the factor using the keypad numbers Default value is 40 range is 0 01 to 9999 Step 7 Press OK to enter the Results screen OR Cancel to return to the Nucleic Acids folder Results Screen Step 8 Insert the reference Press GE Key This will be used for all subsequent samples until changed Step 9 Insert sample and press This measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 absorbencies are calculated both corrected by the background wavelength value if selected Gives concentration based on abs
72. orbance at wavelength 1 Repeat step 9 for all samples Press to return to the Nucleic acid folder Press lt gt to display available Options which are described below Version 5 4 18 4 2011 Page 34 Picodrop Ltd Options select using key pad numbers Eh Parameters 1 Return to parameters screen step 1 above O Prin 2 Print result via selected method O suo 3 Toggle graph on off The graph shows a wavescan plot across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if background correction selected 320 nm 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off E Sample Mumber E Save Method tu Anuto Print RNA Exit options by pressing or wait Version 5 4 18 4 2011 Page 35 Picodrop Ltd 3 Oligo The procedure is as follows Step 1 Press 3 to select Oligo mode Step 2 Select path length using the left and right arrows Options are 1mm tip or 10 mm cuvette Press the down arrow Step 3 dilution factor known Oligo Parameters Pathlength Units Dilution Factor Pore Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace and clear the last digit entered Background OR
73. ows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 Press to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Page 42 Picodrop Ltd BLA FER SHER Calibration Manual entry Shows previously entered calibration values and allows values to Standards T be entered via the keypad 0 051 A de The highlighted box can be edited in order to enter an 01714 A Y absorbance value corresponding to a given concentration value 0 288 A A Jaca using the keypad numbers Range 0 001 to 9999 Use C to Aa vA backspace and clear the last digit entered and the up and down 0 4 0 516 A P arrows to move between boxes 0 5274 BE get a alice socio 2 ah ejoj joejolo Press OK to accept the calibration and go to the Results n a 0 4 E 0 8 10 12 2 4 screen see below OR Press Back to return to the Standards screen Results screen Step 14 Insert the reference and press the GE key This will be used for all subsequent samples until changed wavelength 562 nm Step 15 Absorbance Insert the sample and press 0 288 A The concentration of the sample is taken and displayed Repeat step 15 for all samples Curve Fit Regression Press to return to the Protein Folder Press lt gt to display available Options which are described below Options select using key pad numbers 1
74. p Ltd ene Switch the instrument on and go to utilities instrument preferences and select the Built Auto Print in printer Printer Version 5 4 18 4 2011 Page 63 Picodrop Ltd Loading changing the printer paper 1 Lift off the paper cover Lock the platen and turn the knob to feed the paper 2 Feed in the paper Sometimes it helps if the platen lock is released 3 Paper gripped 4 Replace Cover Version 5 4 18 4 2011 Page 64 Picodrop Ltd Changing to the Bluetooth Accessory 1 REMOVE THE POWER CABLE FROM THE INSTRUMENT Turn the instrument over and remove the cap head screws from positions A and B using the Allen key provided 2 Turn the instrument back over and lift the accessory cover vertically upwards to remove Remove the tie wrap from the cable 3 Plug the accessory cable into the Bluetooth module 4 Note the slots in the base of the case The two lugs on the Bluetooth module plug into these Version 5 4 18 4 2011 Page 65 Picodrop Ltd 5 Note the slots in the accessory cover designed to engage with the Bluetooth accessory PCB 6 Lower the accessory cover vertically downwards onto the instrument engaging the PCB in the slots 7 Invert the instrument and replace the cap head screws at A and B Version 5 4 18 4 2011 Page 66 Picodrop Ltd F 9 Switch the instrument on and go to the rinter preferences page under utilities instrument and select the Printer 4C
75. press This measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 absorbencies are calculated both corrected by the background wavelength value if selected Gives concentration based on absorbance at wavelength 1 Repeat step 9 for all samples Press to return to the Nucleic acid folder Press lt gt to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method O A 3 Toggle graph on off The graph shows a wavescan plot G Frin across the range 220 nm to 320 nm with cursors denoting cramh 230 260 280 and if background correction selected 320 nm 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value Sample Number 8 Save method use the left and right arrows to select a folder EY Save Method to store in Favourites Methods 1 9 press the down arrow El Auto Print and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 37 Picodrop Ltd Protein Determination Protein Determination at 280 nm Protein can be determined in the near UV at 280 nm due to absorption by tyrosine tryptophan and phenylalanine amino acids Abs 280 varies greatly for different proteins due to their amino acid content and consequently the specific a
76. r add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favourites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 52 Picodrop Ltd Bacterial Cell Culture Measurement OD600 Bacterial cell cultures are routinely grown until the absorbance at 600 nm known as OD600 reaches approximately 0 4 prior to induction or harvesting A linear relationship exists between cell number density and OD 600 up to approx 0 6 It is important to note that for turbid samples such as cell cultures the absorbance measured is due to light scattering and not the result of molecular absorption The amount of scatter is affected by the optics of the system distance between the cell holder and instrument exit slit geometry of this slit and the monochromator optics Different spectrophotometer types therefore give different responses for the same turbid sample to compare results they must be normalised using calibration curves A calibration curve can be determined by comparing measured OD 600 to expected OD 600 Expected OD 600 is determined by counting cell number using an alternative technique for example microscope slide method and converting to OD 600 using the rule of thumb that 1 OD 600 8 x 10 cells ml for E Coli Your Piccodrop instrument h
77. rint 3 Toggles on off displaying a graph of wavescan 20 nm Graph from selected wavelength Run Standard 4 Return to Run Standard screen 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value E Sample Number 8 Save method use the left and right arrows to select a folder E Save Method to store in Favourites Methods 1 9 press the down arrow E Auto Print wf and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 5 4 18 4 2011 Page 17 Picodrop Ltd 3 Wavescan An absorption spectrum can be obtained from your instrument enabling simple identification of peak height and position The procedure is as follows Wavescan Parameters Start wavelength End wavelength Mode E OF 0 30 0 25 0 20 0 15 0 10 0 05 400 420 440 460 480 500 a nz as asjaj abs 0 190 0 308 0 088 014 T Sample 1 a 446nm Abs 0 345 4 Version 5 4 18 4 2011 Step 1 Set start wavelength by using keypad numbers or left and right arrows Press the down arrow key Step 2 Set end wavelength by using keypad numbers or left and right arrows Press the down arrow key Step 3 select the mode Absorbance or T using the left and right arrows Step 4 To enter the measurements screen with the selected parameters press OK OR Cancel the selections and return to the Applications Folder by
78. row Step 5 Enter co efficient 1 280 nm using the keypad numbers Default value is 1 55 range is 1 00 to 9999 Press the down arrow Step 6 Enter co efficient 2 260 nm using the keypad numbers Default value is 0 76 Range is 1 00 to 9999 Press the down arrow Step 7 Select the units of measurement using the left and right arrows Options ug ml ng ul and ug pl Step 8 Press OK D to enter the Results screen OR Cancel JM return to the Protein folder Version 5 4 18 4 2011 Page 39 Picodrop Ltd Results Screen Step 9 Insert the reference Press GE Key This will be used for all subsequent samples until changed Protein Uv Step 10 Insert sample and press DJ This measures at both 260 and 280 nm wavelengths and displays the result Protein concentration is calculated corrected by background wavelength value if selected Repeat step 10 for all samples Press to return to the Protein folder Press C Options d to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method Farameters 3 Toggle graph on off The graph shows a wavescan plot EP Frin across the range 250 nm to 330 nm with cursors denoting El Graph 230 260 280 and if background correction selected 320 nm 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value
79. rs long To access a list of pre defined units press the Options key and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters or Cancel Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin interpolated or cubic spline Press the down arrow Step 6 Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates using the left and right arrows This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2 0r 3 Step 8 Press Next O to enter the Standards screen OR Press Cancel to cancel selections and return to the Protein folder Standards Screen Step 9 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears
80. rsors on and off These La 42 996 ass aer show vertical dashed lines displaying the measured peak height Abs 0 282 0 517 0131 0210 1 and horizontal dashed lines showing the peak width Sample 2 A 496nom Abs 0 020 4 Pressing Cancel ignores the selection pressing accepts them Version 5 4 18 4 2011 Page 19 Picodrop Ltd af aVesScan User Defined Peak a T Farameters Print Abs ST Delete Peak Graph Scale Save Method A ubo Frint 0009000009 Graph Scale zoom Mode HW ares H akis limits Peak Detection Sample Mumber A 4396nm Abs 0 020 A Wan escan User Defined Feak 460 480 Soo A496nm Abs 0 020 4 1 EEG 020 3 aby Version 5 4 18 4 2011 Page 20 Add Peak Shortcut button 5 Adds a used defined peak at the current cursor position The entry is then displayed in inverse colouring to discriminate between user defined peaks and auto detect peaks When the cursor is positioned over the user defined peak a legend User Defined Peak appears at the top of the graph The option then changes to Delete Peak to enable the user to remove the peak Note Storing a method at this stage will save these user defined wavelengths each time method is run Absorbance value at these wavelengths is reported Graph Scale This enables the user to set up a defined graph by defining the limits in either or both of the x an
81. s will be used for all subsequent samples until changed Step 12 Press to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press to measure the standard and store the result Bradford Calibration Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading ge 0 2 A Da 04 05 08 LO LE LY Step 13 Press to accept the calibration and go to the Results screen see below OR Press Back to return to the Standards screen Version 5 4 18 4 2011 Page 45 Picodrop Ltd Bradford Calibration Standards 0 20 0 058 A 0 40 0 196 A 0 60 0 330 A 0 830 0 463 A ae 0 4 0 7194 ma DL p d i D2 04 0 56 0 8 LO La 14 Bradford wavelength 535 nm 1 0 3371 4 Curve Fit Regression Farameters Print Graph OOo Sample Humber Save Method Anto Print DoS Version 5 4 18 4 2011 Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Range 0 001 to 9999 Use C to backspace and clear the last digit entered and t
82. t fit of the data points are displayed They may not be relevant for simple kinetics experiments The procedure to define a new method is as follows Kinetics Parameter 1 Screen Step 1 Wavelength Ines E stameters 2 Enter all numerical values using the keypad numbers or the left and right arrows Use the up and down arrow keys to move tice Ui between boxes Step 2 Delay time Enter the delay time in seconds before measurements are taken This can be a maximum of 600 seconds 10 minutes a Step 3 Duration Enter the time in minutes over which measurements are taken This can be a maximum of 60 minutes Step 4 Interval Enter the interval time in seconds between measurements using wavelength Duration Interval 10 Seconds the left and right arrows Options are 5 10 20 30 or 60 seconds Step 5 Press Next to go to the next parameters screen OR Kinetics Parameters 2 Press Cancel to return to the Applications Folder Mode Delta A Kinetics Parameters 2 Screen Step 6 Select the measurement mode using the left and right arrows Partir Delta A change in absorbance over the measurement duration or selected period Final A absorbance at the end of the measurement duration or e o Bak selected time Slope rate of change of absorbance over the measurement duration or selected period Version 5 4 18 4 2011 Page 21 Picodrop Ltd Kinetics Parameters 2 Step 7 Units The user can
83. tings Version 5 4 18 4 2011 Page 60 Picodrop Ltd 2 Su Doku Can be set up as Computer mode 50 preset games or User mode enter your own pattern Use the cursors to select the square and the key pad to enter a number Invalid numbers cannot be entered Cells can be locked or unlocked by using the decimal point Unlocked cells can be cleared using the C key see also option key below The user mode starts with a blank grid Sudoku Setup Sudoku Game 1 Mode Computer Options Press Cc to display the options menu Return to the set up screen The instrument solves the game for you Clear all entries Save the game Use the left and right arrows to select a folder to store the game in Favourites Methods 1 9 press the down arrow and enter name Se Press Cancel to return to the Utilities folder Version 5 4 18 4 2011 Page 61 Picodrop Ltd ACCESSORIES INSTALLATION Printer installation Version 5 4 18 4 2011 Page 62 1 REMOVE THE POWER CABLE FROM THE INSTRUMENT Turn the instrument over and remove cap head screws from positions A and B using the Allen key provided 2 Turn the instrument back over and lift the accessory cover vertically upwards to remove Remove the tie wrap from the cable 3 Invert the instrument and replace the cap head screws at A and B 4 Plug the accessory cable into the printer en Lower the printer onto the locating bosses and push down firmly Picodro
84. ulti instrument lab environment Back up all methods as detailed in section 3 4 Take the SD Card from the instrument and load into a PC with SD memory card Reader Use Windows Explorer to locate the Serial no BACKUP file for the instrument you have just backed up Rename Serial no BACKUP to instrument type BACKUP where instrument type depends upon the instrument you are using e This card can then be restored into any instrument of the same type as per section 3 5 Version 5 4 18 4 2011 Page 75 Picodrop Ltd PRINT VIA COMPUTER PVC Print Via Computer is a small application running under Windows 2000 Windows XP Vista or Windows 7 to enable a Pico200 to transfer data into a PC environment From there the user has a selection of choices the data can be both printed or saved in a variety of formats PVC is capable of supporting several instruments simultaneously limited only by hardware and the speed of the host system PVC can operate via USB and Bluetooth simultaneously PVC can store data either to a common directory or be configured to save to independent directories by both file format and connection PVC can save data in graphics format text format or as an Excel file Installation See the manual included on the PVC CDROM for installation and operating instructions Version 5 4 18 4 2011 Page 76 Picodrop Ltd ACCESSORIES USB cable source locally Built in printer accessory 80 3003 84 Bluetooth accessory 80 3003 9
85. umber of standards to be entered and enter 0 00 for concentration use this when required to enter standard 1 A linear regression analysis of the calibration standard data points is calculated the result together with the correlation coefficient can be printed out A correlation coefficient of between 0 95 and 1 00 indicates a good straight line Version 5 4 18 4 2011 Page 38 Picodrop Ltd 1 Protein UV This is the Christian and Warburg assay discussed previously The procedure is as follows Protein UW Parameters Step 1 Press 1 to select Protein UV mode Pathlength Coeff 1 Step 2 Select path length using the left and right arrows Options are 1mm tip or 10 mm cuvette Dilution Factor Coeff 2 Press the down arrow Step 3 dilution factor known Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace and clear the last digit a entered OR Step 3 calculate dilution factor Press to enter the dilution factor screen Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press O to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down ar
86. ved if a cable is uncoiled incorrectly and damage or weakening can be the result and this type of handling will void the warranty Pulling Do not pull fibre cables by their end fittings And be careful of pulling in general unless your specific cable has been rated to withstand this stress Fibres are usually very strong in direct tension relative to the cross section but when fibres are small it is very easy to break them Other Handling Comments Optical fibre is not wire or rope and can t be handled as such Any fibre optic cable constructed with quality techniques and materials should survive as long as care is taken when handling it The most precarious moments in a fibre cables life occur during inspection testing and installation It is very easy to exceed the bend radius guideline especially when working with large core cables and performing these tasks Version 5 4 18 4 2011 Page 80 Picodrop Ltd
87. volumes with a high degree of accuracy and precision Samples are contained within patented UVpette pipette tips There is no possibility for cross contamination or carry over on a sample platform Precious samples can be handled within a sterile environment and are completely recoverable A 2 5ul sample is drawn up directly into the UVpette using a P10 pipette The pipette is placed into the holder which positions the tip through a light beam emitted from a fibre optic cable connected to the tip holder The light source is a pulsed xenon lamp and the light path through the tip is 0 9mm The default path length is set in all modes to Picotip 0 9mm which is designed for use with UVpette tips as supplied by Picodrop Ltd Resulting data displayed in reports and graphs are calculated to display the equivalent 10mm absorbance values ONLY USE PIPETTE TIPS MANUFACTURED BY PICODROP LIMITED Consumable re order details Ref UVTIPB P10 UVpette pipette 96 boxed tips UV transparent to 230nm Ref UVTIPG P10 UVpette pipette tips 2 000 loose bagged UV transparent to 230nm The user interface is built around folders which are displayed on the home page when the instrument is switched on After switch on and calibration the default home page is Pico200 offering the choice of Applications General spectroscopic methods Favourites A folder to store your more frequently used configured methods Methods Contains nine folders that can store less freque

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