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DropSense User Manual

1. A The user can also browse the available standard curves by clicking the view std curves button This opens a window with all standard curves available for use View Standard Curves View stored Standard Curves linear y Saved standard curves Method linear A562 4 79 C 0 00326 R 0 9880 10mm Absorbance 562nm Concentration g l linear y SELECT C Once the user selected a pre defined standard curve it is not possible to add new ie standard reference samples in the experiment 33 Samples definition Input via graphical user interface trinean Plate Layout EXPORT IMPORT PLATE LAYOUT PLATE LAYOUT nan i used for all unknown samples in the experiment It is not possible to define multiple standard curves in one experiment e An experiment can contain multiple plates the reference samples can be on any location on one or multiple plates e The reference samples can be defined on any plate inside the experiment However if the measurement is stopped by customer interaction instrument failure power breakdown etc before the reference samples are measured no concentrations will be calculated K e There is one set of reference samples to define one standard curve which is al It is not possible to define extra standard curve samples if a pre defined curve is selected It is recommended to define replicates of the references samples to improve the overall accuracy of the
2. Gewijzigd op Gisteren 9 augustus 2011 18 01 DISPOSABLE Disposable type id Name type fial ie eee 5 Number of disposables Dispol DropPlate 1 MATERIAL Group name piat Acids Material name RNA PUMP PROFILE pumping meas pump 14 Can I correct errors in definition of samples after the measurement No this cannot be done in the DropQuant software it is not possible to edit the experiment definition afterwards type of plate type of material position of the blanks etc have several red labeled results Reasons for this may be various e user errors during pipetting e hardware problems on the pumps or manifolds e viscous samples use viscous mode Get the measurement open it and check the pump profiles in the debug tool 15 Spectrum looks skewed part of the spectrum is negative This occurs when a labeled DNA sample is measured without selecting the label in the experiment definition Similar situations occur then for example potassium dichromate using DNA mode Solution is straightforward set the background to no background or single wavelength background correction selecting a wavelength at which sample is not absorbing Theory A DNA RNA concentration and purity determination Nucleic acid samples can be checked for concentration and quality using the DropSense96 spectrophotometer Nucleic acids have an absorption maximum at 260 nm A260 and the spectrographic reading at
3. minimal functionality can perform measurements no password required e User this is the default user level offering the full functionality of the software to start measurements and analyze the results including using formats e Administrator in older versions lab manager has more rights for changing the settings such as the user database and the location of the measurement database Adding more administrator level users requires service login e Service login level for servicing and troubleshooting the DropSense instrument This level requires a service dongle available to trained service engineers only Guest guest None User user1 None Administrator administrator JC7uD7E default password Service service Non available for users At startup a number of default users are created It is recommended to add more administrator level users during the installation process and to change the default password Don t forget to write it down on a secure place The user is the owner of his own measurements this has an impact on the visibility of the experiments By default experiments are visible only to the owner of the measurement and are thus not visible to other users This protects the results of the experiment There are two exceptions e Users can always open and analyze measurements owned by a user of a lower level user For example everyone can analyze measurements done by guest service engineers can open an
4. Set default location Import and export settings The DropQuant program stores a number of user settings in ini files When installing the DropQuant software on a new PC the user needs to transfer the settings from the old PC to the new PC The settings and tool section includes tools to transfer these settings K The import window also allows importing a disposable database This in case IN Trinean updates the disposable database for example when new types of disposables are launched As the user cannot edit the disposable database there is no method to export that information The edit formats settings window The use of formats is not available for all DropQuant users Each user defines his formats and these are by default not visible to other DropQuant users However the administrator can make one or more of his formats visible to all DropQuant users License Some software extensions to the DropQuant software require a USB dongle and activation code The dongle and code are shipped when ordering the software extensions such as the API of cDrop software DropQuant stores the activation code information so that the user doesn t need to input the activation code each time the software add on is started inserting the dongle is always required The DropQuant can store multiple activation codes and auto detects if one of the activation codes matches the dongle s inserted 55 Data viewer extra functions trinean DropQ
5. trinean DropSense 96 and DropQuant v1 5 User manual June 2015 Contents Contents Trinean warranty statement Introduction Warning Safety Information Electrical safety Biological safety Chemical safety Mechanical Specs for DS96 Chapter 1 DropSense96 at a glance DropSense96 Overview Consumables External Characteristics Chapter 2 Software DropQuant Computer requirements Installation Updates Login Home screen New Measurement Plate Layout Import Measurement Chapter 3 Applications Nucleic Acids Oligo measurements Proteins General UV Vis and Standard Curve Standard curves Chapter 4 Interpreting data DropQuant DataViewer DataViewer results window 96 Well Display Column Display Single Well Display Report Display Chapter 5 Exporting data Export Results Export Outliers Export Normalization O W COND AD WI N O 11 11 12 i 19 19 19 19 19 20 21 22 24 26 2 27 28 29 31 3l 38 38 os 40 40 41 42 44 44 45 45 Quick Export 47 Chapter 6 Software extra functionalities 48 Users 48 Bar code scanner 48 Available wells database 51 Chapter 7 Software advanced settings 52 The system info and settings window 52 System info 52 Edit Settings 53 Data viewer extra functions 56 Chapter 8 Diagnostic and calibration tools 57 Optical calibration 57 Potassium dichromate check 58 Quick Check 59 Generate status zip file 60 Chapter 9 Troubleshoo
6. 5500 Trp 1490 Tyr 125 Cys e the extinction coefficient number of Trp Tryptophan Tyr Tyrosines Cys Cysteines A very rough protein concentration can be obtained by making the assumption that the protein sample has an extinction coefficient of 1 so 1 OD 1 mg ml protein In combination with the DropPlate D consumable the DropSense96 performs a dual path measurement with path lengths of 0 7 and 0 1mm to enable quantification of proteins with a wide concentration range without the need for dilution Using the DropPlate S with a single micro cuvette 0 5mm path on the DropSense96 is ideal for quick analysis of protein samples within a more limited measurement range The absorbance data is archived on the database and displayed on the software screen after measurement In the protein A280 application the software chooses automatically which path length generates the best absorbance values and normalizes this measurement to a 1 0 cm 10 0 mm path 66 K Since the UV absorption of nucleic acids at 280 nm can be as much as 10 times that of a4 a protein a small percent of nucleic acids in the sample can greatly distort the protein quantification Therefore the protein sample purity must be determined using the A260 A280 ratio An A260 A280 ratio lt 1 indicates pure protein whereas a higher value indicates nucleic acid contamination An alternative method for protein quantification is a colorimetric protein ass
7. Blank average of blanks Name 6 A260 Concentration 164 48 ng ul A230 10mm 1 43 A260 10mm 3 29 A280 10mm 1 76 Raw A340 10mm 0 03 A260 A230 2 30 A260 A280 1 87 subtract 41 Input Well Selection as it is possible to select a single well to be visualized on this window it s also possible to select multiple wells using either the Ctrl or the Shift key Results REPORT DISPLAY SINGLE WELL DISPLAY COLUMN DISPLAY l 96 WELL DISPLAY Select InputWell show cursor OD 10mm autoscale 0 2mg ml 3 Concentration mg ml G1 0 4mg ml 20 B1 3 06 mg ml HL f PBS D1 5 98 mg ml a2 PBS B2 F1 11 87 mg ml 3 SMENT Fi 11 87mg ml H1 22 11 mg ml fez omeji 2 J 0 1mg ml B2 1 54 mg ml G 0 2mg ml D2 3 11 mg ml A3 50mg ml F2 6 04 mg ml OECD EERE T SS EEE eee POLI 2 blank U E I AEG 230 300 320 340 360 380 subtract ERE EERE Wavelenath EEEE B OOOO background correction OCC OLC EMERMA QUICK EXPORT TO XLS aii W single point 340 nm 10mm Absorbance BACK TO DATABASE EXPORT Blank When using a blank sample it is automatically subtracted from the sample absorbance before concentration calculation This feature can be turned off so the spectral shape and concentration calculation is shown without blank compensation In this mode the shape of the blank spectrum itself can be analyzed When the blank subtraction is turned off all other screens w
8. Performing wavelength calibration hardware parameters successfully updated report saved at Potassium dichromate check DropQuant software includes a performance validation procedure as an easy check up of the performances of the DropSense96 The check is based on the measurement of an aqueous potassium dichromate K2Cr207 solution with a verified concentration Trinean product number 301010 It is good practice to check the instrument s performance every four months depending on the amount of samples measured with a fresh vial of K2Cr207 3 Press Potassium Dichromate test button a new screen pops up with Potassium dichromate automatically entered as sample material 4 Enter the DropPlate of choice DropPlate D or DropPlate S and the target absorbance of the solution given by the manufacturer normalized for a 1mm path length this information can be found on the vial lable The DropQuant software will automatically calculate the expected OD values for all the path lengths used in the DropPlate 5 Load the blank and potassium dichromate solution on a DropPlate as indicated on the screen Use MQ water for this purpose The report display the DropQuant software will show the results of the validation procedure All OD values measured by the 4 optical systems in the DropSense96 are shown 58 For every optical system the mean of all measurements are calculated and compared with the expected OD value of
9. Thank you for purchasing the DropSense96 This is a unique system for the quantification of microliter samples consisting of DropPlate Disposables a DropSense96 instrument and DropQuant software This manual is intended for all DropSense users and it describes the installation operation and measurement procedures An electronic copy of this manual can be opened from the DropQuant program by clicking the Help button Warning Situations that can damage the instrument or your experiment Caution Situations that can potentially affect the results of your measurements K Note Notes usage tips or additional information Warning The DropSense system is designed for UV Vis quantification of small samples Using the system for other applications doesn t guarantee the user s safety and proper functioning of the instrument This instrument is intended to be used by persons having a basic knowledge of UV Vis spectroscopy and the handling of nucleic acids and protein samples Before starting the installation or use of the system read this manual carefully and make sure to understand the instructions to avoid safety issues Keep the manual close to the instrument for reference The content of this manual is subject to change without prior notice Safety Information Before using the DropSense 96 system it is essential that you read this user manual carefully and pay particular attention to the safety information The instructions a
10. The input well is connected to a meander shaped storage reservoir that uses capillary force to take up the sample from the input well This technique protects the samples from evaporation Nr of measurement reservoirs and path length 1 0 5mm 2 0 1 and 0 7mm Recommended sample quantity 2uL 2uL Accepted Variation Volume 30 20 Sample residence time 2h 2h Measurement range OD10mm 0 1 to 40 OD10mm 0 1 to 200 OD10mm Measurement reproducibility lt 2 OD 40 03 OD lt 2 OD 0 03 OD gt 2 ODE t 1 5 gt 2 OD 15 Measurement time for DP96 plate lt 5 minutes lt 8 minutes Software and firmware requirements All systems DropQuant 1 2 and DropSense 14 with firmware version 1 51 The DropPlate D The DropPlate D Dual New Generation has two measurement reservoirs for optical analysis The capillary storage chamber can store up to 2 ul sample By sequentially filling both optical chambers a dual path length measurement is done The combination of both path lengths results in a large OD measurement range and thereby avoids the need of sample dilution Meander shaped Input well reservoir Measurement wells Empty measurement Small path measurement Long path measurement y kes Single chamber measurement Dual chamber measurement The D plate has 2 measurement reservoirs allowing measurements at 0 1mm and 0 7mm path length offering a wider measurement range for higher concentration samples Sample size requirements For c
11. the customer can select to use the internal barcode scanner or not When this option is selected the DropQuant software will automatically read the barcode when the disposable is loaded and check if this agrees with the plate definition or add the barcode to the plate definition When defining the plates in the experiment the user must input the barcode label as name for the plate There are 3 options e Input via the keyboard double click on the default name and edit the name 49 e The internal barcode scanner click on the scan bar code button and the code will be scanned and automatically filled in the table If there is no plate in the instrument the tray opens automatically and the customer can load the plate e Use an external barcode scanner connected to the PC This is explained in the next paragraph trinean Plate Layout 120126w451689 Vie o0000000000 B8 f maY may a mar f ear o prp00r0r000 ppp 0 0t0000 20000000000 pp00 0 0000 prr p r0r00 orppprrngnan CANCEL OPEN TRAY naas lt PREVIOUS START TEST gt PLATE LAYOUT PLATE LAYOUT In case the barcode labels are not known at the definition of the experiment the customer can use the default names During the measurements the program will scan the code detect that the name does not correspond to the plate names defined in the experiment and ask the customer to overwrite the plate name with the code scanned Note that in this ca
12. check starts automatically when activated and takes about 30 seconds The Quick system check consists of 1 Spectrophotometer check the lamp spectrum is measured and compared to the lamp spectrum at calibration The report summarizes the measured amplitude and position of the wavelength peaks and gives a warning if these parameters deviate too much from the calibration values This indicates that the lamp or spectrophotometer may need service 2 Pump system check the output pressure of the pumps is measured using the internal pressure sensors and compared to the values at the calibration This allows checking for degradation of the pumps 3 Motor check the function of the motors and limit switches is checked 4 The operation of the internal background light sensor and the internal temperature sensor are checked After the check a pop up window shows the most important results An Excel report can be created using the create report button This report can be saved for future reference Upon instrument start up or account log in the DropSense96 automatically performs diagnostic checks to ensure that it is functioning within specifications When one of the parameters is out 39 of specification a warning screen appears Please contact your local distributor for a DropSense96 recalibration Generate status zip file This tool allows the user to zip and save a detailed hardware report Please keep in mind that this report
13. e Concentration the calculated concentration The syntax depends on the unit defined in the experiment definition phase For example if unit is ng uL then the name of the column should be Concentration ng uL Select Select elements to Export Export Results Available columns Selected columns Export as Source Plate ID Sample name Include Summary Conientranon Include 96 well plate layout A540 10mm DropPlate Position O xls open in existing Excel file v Pump pdf Raw A340 10mm Material direct print Date Time R square Preview name Concentration EXPORT RESULTS Typical applications The applications for the standard curve include the concentration measurement of proteins with unknown extinction coefficient readout of colorimetric assays etc 37 Interpreting data 4 DropQuant DataViewer To analyze previous measurements we have available a DropQuant DataViewer Search field filter trinean DropQuant DataViewer ID Date User Name Description S d 2010 10 08 Guest Experiment 2010 10 08 16 11 Purified Proteins BSA dilution series ave 2010 10 08 Guest Experiment 2010 10 08 12 14 General UV vis Fluorescein dilution series experiments 2010 10 08 Guest K2CrO7 2010 10 08 11 07 Potassium dichromate test Change user ZIP selected experiment Delete selected experiment Import Database REFRESH NEW SEARCH OPE
14. have an expiration date Don t use the DropPlate after that date as the self filling function is no longer guaranteed During the measurement the instrument simultaneously measures the UV Vis absorption through the measurement cuvettes during the pressure driven transport of the samples from the capillary storage channel to the measurement cuvettes This allows the instrument to monitor the filling behavior of the measurement cuvettes and analyze the spectral absorbance of the sample The microfluidic chip allows accurate and reproducible measurements due to fixed path lengths and elimination of sample evaporation in the input well There are 2 different approaches for our disposables organized in 2 different types of chips The plates 2 different approaches e DropPlate16 this small version of the DropPlate consumable has 16 input wells The DropPlate16 must be mounted on an aluminum carrier the DropFrame Up to 6 DropPlates can be mounted on a DropFrame allowing measuring 1 to 96 samples simultaneously The DropFrame has the same outer dimensions as a standard 96well plate assuring compatibility with liquid handling robots e DropPlate96 this 96well microtiter plate is designed for high throughput use This consumable is ideal for automated workflows as it is easy stackable and it includes a bar code for sample tracking The DropPlate96 is compatible with liquid handling robots SBS standards and it allows easy sample loading pr
15. installed Trinean software and the instrument connected to the PC a Settings trinean System Info and Settings Windows 7 Professional 7 T Trinean Support 5 exe 24 11 2014 FT245R USB FIFO AS00emlW 1432940281 oD e N So pa J z e P Move to safe position Diagnostics Status Check Chapter 8 Diagnostic tools IQOQ On the screen above you can find a Create IQOQ report The software automates the IQOQ as much as possible guiding the user through the validation and creating a report After pressing the button a screen will popup asking for the IQOQ code that can be purchased at your local distributor 52 trinean System Info and Settings The settings of the software are stored in a number of different ini files The administrator user can edit export and import these settings to keep the system up to date Accounts The edit user database allows adding new users and editing the settings of current users Only administrator level users can create new users The only information required is the user name the password and the option to give the new user access to all experiments When the access to all experiments is selected that user can open all experiments by user guest user and administrator levels Also reports can be created from these experiments Service level is required to create new administrator users Contact your technical support Mea
16. measurement and compares the optical transmission of the filled reservoir with the transmission of the empty reservoir This process fails if the reservoirs are already filled at the beginning of the measurement process 4 Can install software on iOS Linux others No software runs on Windows systems only 5 What are operating conditions temperature humidity Operating temperature range is 15 35deg C room temperature this is 59 95 deg F Humidity range is 10 80 no condensation The shipment weight of an instrument is about 30kg Power consumption the external power supply is a SOW type 24V 2 1A 6 Can TRINEAN provide a qualified and documented System Trinean has an add on software product which automates the IQOQ procedure as much as possible It guides the user through the validation process and creates a report The software allows validation of the system for next applications e Qualify the installed DropSense instrument and DropQuant software as generic UV vis spectrometer system for microliter sample quantification e Qualify the installed DropSense instrument with internal barcode scanner and DropQuant software as generic UV vis spectrometer system for microliter sample quantification The validation process includes e Check installation of instrument and software e Check self test of instrument e Check OD performance using potassium dichromate test 62 e Check of the pump profiles is part of the OD pe
17. method This is highly recommended if the OD values of the references are at the limits of the detection range 0 1 OD 34 Sample definition The import section is extended allowing definition of standard curve measurements There is one extra column for the definition of the reference value If this column contains a number then the sample is by default a reference If it is empty then this well is an unknown sample trinean Import Samples xis xlsx csv y Import format Default Nr of rows to skip ma header not included 5 S Blanking information Multiple blanks header F DropPlate ID column 3 Source Plate ID column 1 Position column 4 DropPlate ID of blank column 6 Source Postion column 2 Sample name f column 5 Blank Position column 7 Dilution factor column f lt not in file gt Reference value column 8 converted sample s selected sample s The settings of the standard curve are part of the format and are thus not part of the import file The results window Standard curve experiments can be selected from the measurement database and opened in the post processing window just as any other DropQuant measurement The results window shows the measured spectra of all samples references and unknown samples The concentration of the unknown samples is calculated and shown in the tables 35 Results Ss trinean Results REPORT DISPLAY SINGLE WELL DISPLAY COLUMN DISPLAY 96 WELL DISPLAY
18. the applicable operating and servicing manual and provided further that the products have not as determined solely by TRINEAN been subject to misuse or abuse by the Customer or other parties unrelated to TRINEAN TRINEAN makes no warranty expressed or implied as to the fitness of any product for any particular purposes other than those purposes described in the applicable operating and servicing manual nor does TRINEAN make any other warranty whether expressed or implied including merchantability other than those appearing on the face hereof Where TRINEAN guarantees any product whether under this Warranty or as a matter of law and there is a breach of such guarantee the Customer s only and exclusive remedy shall be the replacement of defective parts and materials as described above This shall be the limit of TRINEANS liability Trinean will provide spare parts of DS series instruments and related items will be made available at least 7 years directly or through third parties e g distributors Furthermore TRINEAN shall not be liable for incidental or consequential damages Failure of the Customer to notify TRINEAN of a claimed defect by registered mail within thirty days of the discovery thereof shall constitute a waiver of any claim for breach of warranty When a product is required by TRINEAN to be installed by a TRINEAN certified engineer the period of this Warranty shall begin on the date of such installation provided howev
19. the potassium dichromate solution Color indications are shown giving a clear overview o Green color indicates that the measurement is within the expected measurement variation The DropSense96 performs within specifications Red color indicates that the measurement does not fall within the expected variation Contact your local distributor for a systems check up A report txt xls odf can be created using the export button This report can be saved for future reference In case of red flags Inaccurate measurements can be caused by 1 The reference vial being open longer than 30 minutes leading to abnormal high OD values 2 Time between dispensing and measuring is longer than 2 hours leading to evaporation 3 Insufficient dispensed volume leading to incorrect filled micro cuvettes 4 Inappropriate blank 5 Obstruction of the optical path large particles finger print small air bubbles 6 The pumps performing out of specifications see color indications on the thumbnail view 7 The optical performance of the DropSense96 is out of specifications 8 When red values are shown repeat the validation test with the opened vial if repeat can be done within 30 minutes from the test or with a fresh vial If the result is consistent with the first test please contact your local service engineer Quick Check This tool performs a quick check of the spectrophotometer module and the pump module in the DropSense96 This quick
20. this wavelength is the most common method for detecting dsDNA ssDNA RNA and oligonucleotides on a solution The nucleic acid concentration is calculated using the Beer Lambert law see below which predicts a linear change in absorbance with concentration Using this equation with a 1 cm path length an A260 reading of 1 0 is equivalent to 50 ng ul dsDNA 33 ng ul or 40 ng ul single stranded RNA Beer Lambert Law A e c A absorbance at a particular wavelength e the extinction coefficient c concentration of nucleic acid path length of the spectrophotometer cuvette The absorbance data is archived on the database and displayed on the software screen after every measurement On the nucleic acid application the software chooses automatically which path length generates the best absorbance values and normalizes this measurement to a 1 0 cm 10 0 mm path These are displayed in the software for further data analysis DNA RNA purity determination Residual cellular contaminants like proteins or compounds used in the DNA RNA preparation frequently remain present in the DNA solution and often interfere with the measurement at 260 nm leading to incorrect results Both protein and DNA absorb UV light but have different absorbance curves The peak of light absorption for DNA is at 260 nm while proteins absorb strongly at 280 nm mainly due to tryptophan and tyrosine side chains Therefore the purity of a DNA sample can be calcu
21. using the spectral range of 400 600 nm or a single wavelength of choice DropPlate type Depending on the application or sample concentration range a choice of DropPlate S or DropPlate D can be made Both DropPlates are explained in Chapter 1 Alternatively a standard 96well microtiter plate can be read This selection requires additional information such as path length sample volume and plate brand Path length selection When selecting the DropPlate D the user can choose between a single or dual path length measurements When using a DropPlate S only a single path length measurement is possible This selection requires a sample volume of 2 ul and generates a measurement range of 0 1 40 OD for a 10mm path Save as Format When all the above selections have been made the user can save these in a new format for future use These formats are username dependent Only the user administrator has the possibility to make a format that is available for all users The formats can also be deleted and renamed Plate Layout After defining the experiment it is time to set up the plate settings On this screen it is possible among other options to set up multiple plates Despite multiple plate selection is possible only ONE 96 well plate can be loaded and measured at a time into the DropSense 96 Loading multiple plates will damage the instrument trinean Plate Layout Plate layout definition Sample name definition table G
22. ASUREMENT NEW DATABASE MEASUREMENT DataViewer Measurement Database Help Define and start anew measurement 20 Setting up a new measurement with DropQuant requires a few steps On the New Measurement window it is possible to define the following settings New measurement es trinean Star new measurement CANCEL Experiment definition Including 2 editable fields the user can define the experiment by name and include extra information on the description box A default name is generated for every experiment and it includes date and time for easy chronologic storing All the experiment information will be included in the final experiment report Sample definition Several pre configured applications have been included for quick and easy experiment definition e Nucleic acids e Purified Proteins e General UV VIS e Standard curve Each of those applications has multiple sub options that will be explained in the Applications chapter of this manual Solvent Despite not having any influence on the measured spectrum this option could influence pumping times of the samples High viscosity In some cases where the samples are rather viscous it might be necessary to activate this mode The high viscosity mode allows the pumps to work for some seconds longer than normal allowing the viscous sample to reach the measurement chambers 21 Background selection Choice between the default background correction
23. AlexaFtuorssa no so O omw oso neeru z0 sooo OO ooo O ooo AlexaFtuorss0 a0 a OO ooo O oo om Too o o y a O oS E a E a S a E G owes owo e O ow o S O G owes o o s O oo O oo G owwa o o s o oo O os x ST a i oms A ne Add Label Save Changes 2 Oligo measurements An oligo is defined by its sequence Its characteristics such as the molecular weight and molar extinction coefficient can be calculated from the sequence and they can be used in the concentration calculation These equations are well described in literature DropQuant allows defining the sequence of each sample and to use this information on the post processing Concentration is expressed as ng uL or pmol uL Conversion between both depends on the molecular weight which can be calculated from the sequence Nucleic acids such as DNA are characterized by a concentration factor which is the concentration as function of the absorption ng uL OD The extinction coefficient is the absorption as function of the mol pmol uL OD the concentration and the mol are related by the molecular weight g mol Defining the sequence 1 On the experiment definition screen select Sample Type gt Nucleic gt Sample Material gt Oligo 2 Select the unit of concentration ug ul or pmol l The selected value will be used on the post processing window Default value is ug ul Other options like solvent labels and background co
24. DNASOug u f ona soue Dna sou DnA 200 blame 32 Dna Sug onAsus CANCEL OPEN TRAY ee ee lt PREVIOUS START TEST gt ee ea PLATE LAYOUT PLATE LAYOUT DropPlate Name Barcode and Description It is possible to set up different plate names for each plate In case of a DropPlate 96 with a barcode it is possible to use an internal or an external barcode reader In case of an internal barcode reader available at Trinean is installed no manual scanning is necessary Scanning a bar code also has the advantage of having the system saving all the available wells on an plate into a database 22 Select click mode These two buttons allow the user to change between the insert mode where it is possible to add new wells or replace existing ones to view update mode where it is possible to navigate through pre defined wells and redefine information such as Well name and Type well Plate layout definition here you can define the plate layout being possible to select several options e Type well Sample Blank Reference for reference curves Note that on sample selection it is possible to define replicates E EJE x Blanking information is also available when selecting samples e Autoblank calculated by software e Blank Average of blanks or specific blank e Well name It will be reflected on the table at the bottom of the screen e Source plate ID In case of pipetting the samples from a stand
25. E DropPlate 1 o __ 19 eo yes os SSS Seas SE DropPlater 0 0 77 BSA o10 10 TP EI Incl lier KM include outliers EXPORT RESULTS W Group by replicates Selected columns All available columns Here it is possible to find several fields with the data available to be exported It is also possible to move the fields up and down in this way organizing the selected columns table above Export Settings It also possible to define some extra settings regarding the final file format It is possible to choose between 4 file types and direct printing to include a summary and the plate layout and some extra information related to the Excel file 44 Export Outliers It is also possible to export outliers from your experiment On this screen it is possible to define what to export and which are the ranges you want to use as a filter trinean Select elements to Export Export Results Export Outliers Export Normalization Export as F Measurement fail Fj Include Summary A260 Concentration E Raw A340 10mm direct print i W A260 A230 inrange pdf E A260 A280 _inrange y open in new Excel file txt ic nel GO GG S S ng ul Propriae 1 semper D SS CC DropPlate 1 DropPlate 1 Preview DropPlatei Samplel DropPlate 1 DropPlate 1 Export Normalization The last tab on the export window allows the customer to calculate the quantities of sample an
26. N DATA Experiment management settings The DataViewer front page shows all the measurements ran on your DropSense 96 It is important to note that some experiments might not be visible to certain users due to access rights For more information please read the Users section in this manual Search field filter To help the user searching for previous measurements based on a specific field it is possible to filter the desired measurements by specifying relevant data Saved Experiments All measured experiments accessible to the current user will be displayed here It is important to while setting up an experiment to input as much information as possible so it can be searched while filtering results 38 Experiment management settings This set of buttons allows the user to export and import experiments or databases e Change user Change the user assigned to that experiment e Delete selected experiment e Zip selected experiment Useful for diagnostics Whenever facing issues with the DropSense 96 it is important to send the experiments where the issue first occurred to your local distributor e Import Database As it is possible to use both DropQuant and DataViewer as standalone software you can also import experiments saved on a different computer DataViewer results window After a measurement two options are possible depending on the selected method on DropQuant s experiment definition As default Dro
27. STANDARD CURVE Select InputWell show cursor A OD 10mm autoscale Concentration 3 10mm Absorbance m e es gie 4 E blank 300 350 400 450 500 550 600 650 700 E subtract Wavelength gt background correction QUICK EXPORT TO XLS H linear W single point 340 nm BACK TO DATABASE EXPORT There is an extra tab showing the standard curve Results trinean Results REPORT DISPLAY SINGLE WELL DISPLAY COLUMN DISPLAY 96 WELL DISPLAY STANDARD CURVE linear y Method linear A540 0 331 C 0 466 R 0 9979 3 amp g Q 2 lt E E S SAVE STANDARD CURVE 30 35 40 45 50 55 60 70 Concentration pg l linear y BACK TO DATABASE EXPORT It is possible to change the method from the post processing window 36 Saving a standard curve for future use On the standard curve tab the user can click the save standard curve button to save the Standard curve for future use The user has to fill in a name for the curve It is important to keep in mind that for some types of experiments a new standard curve should always be used instead of a previously saved one Please enter the name Cancel Exporting the results The export data function includes a few extra columns in case of a standard curve experiment e A540 10mm if 540 is the wavelength chosen for building the standard curve the measured OD used for creating the standard curve
28. TODO ETODO TODO FTooo roDo sroco DONE Busy TODO TODO FToDo FroDo TODO TODO FroDo Frove Frovo Froco DONE Busy TODO FTODO FToDo FroDo TODO TODO TODO FTODo FroDo FTovo DONE susy TODORTODCOIRTODOITODO Measuring DropPlate 1 1 of 1 Instrument nr 2012 0178 2013 12 02 10 15 ease remove DropPlate from tray Applications 3 There are some important differences and extra options when selecting the 4 available Sample Types For extra information about DNA RNA Protein concentration and purity determination as well as General UV Vis spectrophotometry please consult Annex 1 Theory Nucleic Acids Labeled Measuring the labeling efficiency of fluorescent tagged probes before micro array hybridization eliminates potentially flawed samples and improves experiment effectiveness The full soectrum analysis with the DropSense96 allows measurement of nucleic acid absorption at 260 nm while detecting the fluorescent dye on their absorption peak In a single measurement the DropQuant software calculates the nucleic acid concentration expressed in ug ul and the dye concentrations in pmol ul Labels Here it is possible to choose between several types of predefined labels Selecting this option activates the Labels database screen where it is possible to add or remove labels Labels Database ae ETEM Nm AlexaFtuorsas awo seo O onoo ow y O S nearuorsss asooo ss ooo ooo
29. ample pnas DropPlate Blank Protocol Plate ID Position ID Position type ID of blank a CANCEL File type selection Select between the available file types It is also possible to select between spreadsheets on a particular file Plate Layout Sample Definition Sheet3 Import format settings After pressing the arrow next to Default it is possible for the user to define the format to be imported Please note that if errors are shown on the information displayed on the converted samples selected samples info they are most likely to be related to something being not correctly defined here Import format Default Nr of rows to skip rz header not included y Blanking information Multiple blanks y save header DropPlate ID e ID save as Source Plate ID Source Plate ID Position DropPlate Position DropPlate ID of blank DropPlate ID of blank Sample name Blank Position Position of blank y Source Postion Source Position Sample name Dilution factor column lt not in file gt cDrop Protocol 1 Source Plate ID 25 Measurement While measuring the plate DropQuant has a succession of windows with plate loading instructions and information that allow the user to see the measurement progression select ina list Please insert the next DropPlate and select from the list DropPlate 1 Cancel trinean TODO ETODO TODo sTroDo roo srono DONE susy TODO TODO FroDo grove ABORT TEST
30. ard 96 well plate to our plates it is possible to add a common identifier such as plate ID or a bar code e Source plate Position In case of pipetting the samples from a standard 96 well plate to our plates it is possible to add a common identifier that allows to track the position of this sample on the plate of origin Open results in for customers with cDrop it is possible to open the results after the measurement immediately with cDrop for content analysis For all the other customers the default setting will be DataViewer This option must be selected before defining the sample locations s on the plate It is also possible to select between multiple protocols by selecting the sample first and defining it on the plate layout and then the protocol Sample name definition table Despite being possible to define a plate layout and the sample name on the menus above it is also possible to define them here using a table format Simply input the desired name s on the table after the sample is defined above It is also possible to copy values from a chart in Excel to this one simply by dragging the table or copy pasting it This is quite useful when working with complex layouts Additional Settings Right clicking on the Plate Layout definition field it is possible to access a set of options that allow the user to fill plate positions in a serial mode 23 Fill from this position down Fill from this position right All 96well
31. art of the experiment Click on new experiment and on the start experiment window select standard curve 31 New measurement 53 trinean Start new measurement lt proportional linear interval 2nd order polynomial 2nd order polynomial through zero sigmoidal CANCEL Next input fields are available to define the parameters of the standard curve method e Sample material for information purposes only required field e nformation e Wavelength the wavelength at which the standard curve is calculated C This cannot be changed by the customer after the measurement is done sf Still keep in mind that the Method can be changed later e Unit the unit of the reference value e Method e Proportional e Linear OD A a e 2 order ae aoni A conc 2 B conc e 2 order Pa nom through zero OD A conc 2 B cor e sigmoid OD A 1 exp Conc B C e 4 parameter curve e fit OD A D 1 conc C B Case 2 use a pre existing standard curve in the experiment It this approach it is assumed the DropQuant user has already performed an experiment with a standard curve before and saved the standard curve of that experiment for future use 32 The standard curves available for selection are directly available when clicking the measure new standard curve button i Standard Curve pass lt Measure new standard curve water based solvents incl TE PBS RT buffer
32. asurements cannot open import a DropFile Can correct errors in definition of samples after the measurement Spectrum looks skewed part of the spectrum is negative 61 1 The disposables expiry date is overtime The expiry date is 12 months increased from 6 to 12 months in Q2 2011 It is not recommended to use the disposables after the expiry date The customer must use the disposable before the end of the month indicated on the expiry date 2 What if too much sample is dispensed DP S DP D Maximal sample quantity 2 2uL 2 5uL There are 2 effects if too much sample is dispensed e the excessive sample volume remains in the input well resulting in evaporation and a change in concentration e there isa risk that in some wells the measurement reservoir gets overfilled during pipetting This overfilling results in bad measurements The probability of overloading is quite small max few wells per 96 well plate but too large to guarantee error free operation of plates when loaded with too much sample volume 3 Can re use a disposable No it is not possible to re use a disposable the risk of cross sample contamination increases and the self loading properties disappear resulting in low yield of the sample pumping process It is neither possible to do a second measurement on the disposable with the same samples The measurement algorithm assumes the sample is in the meander and the measurement reservoir is empty at the start of the
33. atically at the cursor position Be sure to set the correct parameters for the scanner type of barcode type of language for the keyboard etc Available wells database In case the user uses DP96 format plates and an internal barcode scanner DropQuant is ready to keep track of the plates and wells used by the system DropQuant includes the software to keep track of the wells used After scanning a bar coded plate DropQuant will immediately lookup this barcode in the used well database and show a cross icon for the wells that are already used K This function is also available for manual barcode scanner operation Limitations e This function is available for DP96 plate only For customers using DP16 format plate on aluminum DropFrame there is no way for unique identification of the wells used e fcustomer uses multiple PCs connected to DropSense then these PCs must share the same available wells database Selecting the used well database is explained in next paragraph Managing the available wells database please check Chapter 7 51 Software advanced settings The system info and settings window Administrator accounts have access to the Systems info and Settings window On the main window there is a button on the left bottom corner This button opens a new window containing two tabs for system info diagnostic tools and software setting options System into The system info window gives information about the
34. ay such as a BCA Bradford and Lowry assay These are commonly used for quantification of protein solutions and cell lysates These types of assays can easily be performed with the DropSense96 using the general UV vis mode in the software Quick export of the data to excel allows fast generation of a standard curve and concentration determination General UV Vis spectrophotometry In the general UV Vis application the DropSense96 functions as a conventional spectrophotometer An absorbance scan from 230 to 750 nm of any liquid samples can be analyzed enabling simple identification of absorption peak heights and positions An unlimited number of wavelengths can be designated in advance for absorbance monitoring and inclusion in the report Note that the values reported are NOT normalized to a 10mm path length so if comparison is being made to other systems the appropriate factor should be used to convert them Principle of standard curve measurements In previous paragraphs the concentration measurements are based on direct quantification the OD10mm spectrum is measured and the concentration is calculated from the OD at a certain wavelength using the concentration factor which is derived from the Beer Lambert law This requires knowledge on the extinction coefficient of the substances The standard curve method offers an alternative approach The starting point is a set of reference sample with known concentration The DropSense syste
35. d It is dependent on both the characteristics of the sample pH ionic strength and the wavelength accuracy of the spectrophotometer used Since the DNA absorption peak shows a steep slope at 280 nm a slight shift in wavelength accuracy 1 nm can result in 0 2 change in the 260 280 ratio Protein concentration and purity determination Protein concentration like nucleic acids can be determined by measuring their UV absorbance at 280nm and calculating the concentration using the extinction coefficient of the protein in the Beer Lambert equation This method is suitable to quantify purified proteins However the 280nm absorption of proteins depends on the presence of aromatic amino acids Trp Tyr and Phe and Cys Cys disulfide bonds The UV absorption of proteins varies greatly and depends on the protein s particular amino acid concentration In addition buffer type ionic strength and pH affect the UV absorption and even pure protein solutions may have different conformations and modifications When performing protein A280 concentration measurements the best approach is to empirically derive the extinction coefficient for the protein of interest or search for published protein extinction coefficients examples in the Practical Handbook of Biochemistry and Molecular Biology Alternatively if the protein sequence of the protein to be measured is known the theoretical molar extinction coefficient can be calculated using the equation e
36. d buffer to be mixed to achieve normalization the goal is to dilute the samples to equalize the concentration of all samples to the same concentration and optionally also same volume 45 trinean Select elements to Export Export Results Export Outliers Export Normalization Export as Sample volume pl Fi Include Summary Concentration Normalize to concentration Eg ng ul Normalize to lowest concentration 225 89 ng ul pdf direct print Volume O xls open in new Excel file End volume Maximum sample volume Preview DropPlate 1 Sample7 dsDNA i q 375 39 Fj Include outliers EXPORT NORMALIZATION _save save asf rename There are two different operation modes Add buffer to the sample to achieve a target concentration It is assumed all samples are available in the same volume For this mode select maximum sample volume Note that normalization cannot be done if the sample concentration is smaller than the target concentration In that case the value of the added buffer volume on the table will appear as a dash To have a solution the normalized concentration must be smaller than the smallest concentration measured in the set of samples DropQuant includes the option to set the target concentration to the smallest concentration of the set of samples In that case the smallest sample needs no dilution Then the added buffer volume should be O Take aliquot of the sample and mix w
37. d analyze measurements done by guest standard user and administrator and so on e tis possible to give a user access to all measurements owned by all other users up to administrator level Bar code scanner The 96 well disposables all have a barcode with a unique label The DropSense instrument can be extended with an internal barcode scanner and the DropQuant software includes a lot of functions related to the barcode 48 The barcode on the DropPlate The type of barcode on the DropPlates is a universal code type CODE 128 2 dots and 14 characters This code can be read by all barcode scanners The syntax of the unique code is ABCCCDDEEFFFFF with A is the product number B is engineering revision number CCC is supplier batch number DD expiry date encodes as the number of weeks since 01 01 2010 EE is the coating run number FFFFF unique disposable number within lot Trinean defines this code there is no possibility to order DropPlates with user defined codes g AINIDI ai ni a DP S WT I TT TT exp date 12 2010 a 000000009000000000 The software reads the barcode and performs a number of checks e Type of the plate should match the experiment definition e Expiration date e Check if the plate has been used before and disable the wells that have been used before The position and size of the barcode on the frame is shown below The experiment definition When defining an experiment
38. despite useful should always be complemented with experiment Tiles 60 Troubleshooting 9 Solving an issue In case of issues with the instrument that are not included on our FAQ list please contact first your local distributor In order to have this process running smoothly please follow these steps Export and zip the experiments when the issue started to happen Include also at least one experiment right before the issue appeared for the first time In case of startup issues please generate a status zip file Write in detail what you were supposed to see on that experiment Some of the issues might be related to a specific assay and not to hardware On our website you can find on the support section a checklist to be sent to the distributor in case of issues FAQ list Please bear in mind that this list is being constantly updated For a full FAQ list please consult our website 1 The disposables expiry date is overtime 2 What if too much sample is dispensed 3 Can I re use a disposable 4 Can linstall software on iOS Linux others 5 What are operating conditions temperature humidity 6 Can TRINEAN provide a qualified and documented System 7 What software add ons are dongle protected 8 Can luse my dongle after an upgrade of the software add on program 9 What if lose the activation code 10 12 13 14 15 What if lose the dongle Can copy the dongle changed measurement database but cannot see the me
39. er that any use of the product prior to such installation shall at sole election of TRINEAN void this Warranty When installation by TRINEAN personnel is not required the period of this Warranty shall begin on the date of shipment from TRINEAN The period of this Warranty shall begin as described above whether or not the product has been installed pursuant to a purchase order and any trial period shall be deducted from the Warranty period that would otherwise apply under a subsequent placed purchase order for that product Limitation of Liability Notwithstanding anything to the contrary contained herein the Seller whether by reason of breach of warranty breach of contract tort or otherwise including without limitation under any indemnification provision contained herein shall be limited to replacement of goods returned to TRINEAN which are shown to TRINEANS reasonable satisfaction to have been nonconforming or to refund the purchase price or if not paid to a credit amount of the purchase price therefore THE FOREGOING WARRANTIES ARE EXCLUSIVE AND A RE GIVEN AND ACCEPTED IN LIEU OF ANY AND ALL OTHER WARRANTIES EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION THE IMPLIED WARRANTY OF MECHANTABILITY AND THE IMPLIED WARRANTY OF FITNESS FOR A PARTICULAR PURPOSE NEITHER PARTY SHALL BE LIABLE TO THE OTHER FOR ANY INCIDENTAL INDIRECT SPECIAL OR CONSEQUENTIAL DAMAGES Warranty statements may differ in specific countries Introduction
40. ers a complete system to the customer including a number of add ons S Contact your local distributor to get more information on these extensions to the DropSense system In order to assure an optimal functioning the DropSense96 uses our proprietary DropPlates described below Consumables The DropSense is the only platform that can read DropPlates a full 96 well plate can be read in less than 5 minutes The Trinean DropPlate consumables are made of advanced technical plastics allowing high optical transmission over the full UV VIS spectrum with a high accuracy The basic parts of the disposable are e The conical shaped input wells are positioned with a 9mm SBS standard pitch fit for multi channel pipettes or robotic sample loading e These input wells are connected with a capillary storage channel When dispensing a sample droplet into the input well it is immediately transferred into the storage channel through capillary forces in order to suppress sample evaporation This allows the user to perform the measurement on the DropSense96 within a time span of 2h after dispensing 12 e The capillary storage channel is further downstream connected with one or two measurement cuvettes for optical readout as shown below Those same cuvettes have a fixed path length e The microfluidic channel continues via a small channel towards the vent hole to which the DropSense96 instrument connects its vacuum pressure system All DropPlates
41. eservation and measurement of the droplet optical absorption C It is not necessary to fill all 16 input wells to perform a measurement A single IS sample measurement can be done and the unused positions on the DropPlates can be filled in another measurement until all input reservoirs have been filled DropPlates are single use disposables Don t try to re use a plate The meander s self filling behavior disappears and pipetting will result in air bubbles 13 The frames Up to 6 DropPlates16 can be placed on a DropFrame creating a full 96well plate with standard 96well plate dimensions SBS standard The DropFrame includes mechanical features defining a unique position of the DropPlates This plate can only be placed in one way on the DropFrame due to the alignment pins and orientation triangles on the frame It is recommended to put the DropPlates16 on the frame before loading the liquid samples The DropFrame is black colored to aid the visual inspection of loaded or empty input wells and associated sample reservoirs This specific design of the DropFrame minimizes human errors during sample loading DropPlate D and DropPlate S 2 different types for your needs Currently Trinean has available 2 types of plates the DropPlates S with one measurement cuvette and faster readout time and the DropPlate D with larger measurement range All DropPlate types have identical external dimensions and conical shaped input wells
42. he instrument C3 Do not use solvents or reagents containing acids alkalis or abrasives to clean the DropSense 96 Damage to the touchscreen and computer C4 Do not pour or spray liquids e g cleaning agents on to the DropSense 96 Use a tissue moistened with water only for cleaning gt gt D We recommend wiping the DropSense 96 with a damp cloth only The following disinfectants and detergents are recommended for cleaning the DropSense 96 Note If cleaning agents different from those recommended are used ensure that their compositions are similar to those described below General cleaning of the DropSense 96 e Mild detergents e 70 ethanol Maintenance General instructions Do not use spray bottles to spray cleaning or disinfectant liquids onto surfaces of the DropSense 96 If solvents or saline acidic or alkaline solutions are spilt on the DropSense 96 wipe the liquid away immediately Follow manufacturer s safety instructions for handling cleaning agents Follow manufacturer s instructions for soaking time and concentration of the cleaning agents exposure for longer that the recommended 69 Glossary X Sample size requirements Although the dispensed sample volume is not critical it is essential that a minimal amount of sample is dispensed for correct filling of the measurement chamber allowing precise measurements Extensive testing indicates that sample volumes of 2 ul are sufficient to ensure reprod
43. ill generate data without blank subtraction C These settings are not stored to the DropFiles next time the DropFile is opened the p data is shown using the original settings at time of defining the experiment It is also important that all blanks contain MQ or similar water only Background correction the background correction method can be selected on the Start new measurement pane while setting up a new experiment The background information is derived from the RAW OD spectrum measured On its a fit through the 400 600 nm region of the transmission spectrum is subtracted from the full soectrum On the mode the background level of a wavelength of choice is subtracted from the full spectrum 42 This window shows a small report including all the necessary information for a classic measurement Here it is also possible to change the experiment description Some extra information such as Measurement date DropPlate type used Performed by among others can also be found on this screen This information might also be important for diagnostic purposes when an issue appears t rin ean Results REPORT DISPLAY SINGLE WELL DISPLAY COLUMN DISPLAY 96 WELL DISPLAY MEASUREMENT DATE TEST PERFORMED BY 2010 10 08 16 12 Guest Name Concentration mg ml x A260 A280 DROPPLATE PATH LENGTH 0 05mg ml 0 04 mg ml DropPlate D Double path length 3 1mg ml 3 06 mg ml EXPERIMENT NAME INSTRUMENT ID 0 1mg ml Experime
44. ion value and unknown samples in the sample definition window The DropSense measures the plate and thus the optical absorption for each sample The software calculates the standard curve this is the relation between measured OD and input reference concentration The user can select the type of curve linear polynomial etc The standard curve is shown as OD f concentration Based on this standard curve the concentration of the other unknown samples is calculated from the measured OD value and the constructed standard curve Thus concentration f OD In the results window the standard curve is shown The standard curve is an analytical equation fitted to the measured OD of the reference samples The user can select from a number of models shown on the table below Standard curve equation N a0 CONC a_i CONC OD o Linear OD a0 al COnC ooo o OD D 2 order polynomial OD a0t a1 CONC a2 CONC 2 Po 2 order polynomial through OD a1l CONC a2 CONC42 0 0 OD a0 1 exp CONC a1 a2 OD A D 1 conc C B D Ooo 68 Maintenance B The following maintenance procedures and performance checks must be carried out to ensure reliable operation of the DropSense 96 Cleaning the DropSense 96 Important Switch the instrument off and disconnect the line power cord from the power outlet before cleaning Risk of electric shock W7 Do not open any panels on the DropSense 96 Damage to t
45. ith buffer to achieve normalized sample with target volume and target concentration The aliquot volume taken from the sample is here identified as the used sample volume the buffer volume to be added is the added buffer volume For this mode set the end volume to the desired value Here there is no solution if the target sample volume is too large and or if the target concentration is too large Once this is selected the required sample and buffer volumes will be automatically calculated 46 Quick Export For the user s convenience DropQuant DataViewer also allows to quickly export the data to xls format Please note that this function is only available on the single well display window and that in order to export more than one measurement well the left column has to have the desired measurements selected DropSense REPORT trinean TIT 2 E E OropP lat ne Faw Semplenseel E oe A220 eee Peet asg laoaead Renn ate ion H mm H mm H mm Position 10mm rigful o 188 DropPlate1 E4 Samplei 0 34991 307 FT 1 88 DropPlate1 F4 Sample6 0 amzer 24 545 0 l oo 0 oo 0 0 m m g T i T h k m DropPlate1 BS Sampled DropPlate DS Samplet h hk k k AZS 402 2 IETT 327 41 i OropFlate 47 Software extra functionalities 6 Users The functionality of the DropQuant software depends on the user level at login e Guest
46. lated by examining the ratio of the two absorbance values A260 A280 values of 1 7 to 1 8 predict clean DNA good quality RNA will have a 260 280 ratio of 1 8 2 0 Lower values may be indicative of significant protein phenol or other aromatic compound contamination However the A260 280 ratio is not always an accurate representation of DNA purity Other contaminating substances like EDTA and carbohydrates have a low 280 nm absorption but absorb UV light around 230nm Furthermore some proteins containing few aromatic residues have little absorbance at 280 nm while all proteins have a clear absorption peak at 228 nm due to their peptide bonds This makes the 260 230 ratio often a more constant indicator of the presence of protein in a nucleic acid sample Therefore absorbance readings measured both at 230 nm and at 280 nm provide a more accurate estimate of contaminants that may be present in nucleic acid samples The ratio of the A260 A230 should be 1 8 or greater since nucleic acids have an absorbance minima at 230 nm 65 K It is recommended to use the absorbance readings at A260 280 and 230 and IS examining both A260 280 and A260 230 ratios for every sample As a general rule a 260 280 ratio of 1 8 and a 260 230 ratio of 2 0 or greater predict clean DNA good quality RNA will have a 260 280 ratio of 1 8 2 0 and an OD ratio 260 230 of 2 0 or greater The 260 280 ratio can differ depending on the spectrophotometer use
47. lows From the measured absorption spectrum the quantity and quality of small biomolecules such as DNA RNA and proteins is calculated This platform is the answer for many applications in life science labs Dropsense96 DropQuant Multi channel reader Measurement software UV VIS spectroscopy Experiment layout Full Soectrum analysis Spectral analysis Easy data export cDrop Analytical software Sample content Specific quantification Quality analysis DropPlate 16 96 Disposable microfluidics micro cuvettes Anti evaporation system The DropSense96 is composed by e 5 channel spectrometer for reading the UV Vis absorption spectra e Pump and manifold unit for that allows the sample to run inside the disposable cuvette e Mechanical stage for moving the disposables inside the instrument The system includes the following measurement modes 1 Direct quantification the concentration of the substances is calculated from the absorption curve using Beer Lambert Law OD e C L With e the material dependent extinction coefficient C the concentration L the path length of the beam passing through the sample and OD the absorption in the sample defined as log P Pref DropQuant includes pre defined values of the extinction coefficient of most frequently used materials 2 groups of biomolecules are supported nucleic acids and purified proteins Based on the selection of the material DropQuant gives more information on the sam
48. m measures the absorption spectra and builds a standard curve this is the relation between the reference concentration and the measured OD at a certain wavelength The samples with unknown concentration are also measured the OD at the reference wavelength is measured and the standard curve is used to calculate the concentration of the unknown sample This method is typically used in case of colorimetric assays where the OD depends on local factors such as temperature pH value and so on These factors are calibrated in the standard curve There are 2 methods to define the standard curve e The samples used to build the standard curve are part of the experiment e A standard curve of a previous measurement is used The customer can save a standard curve for future use when opening a standard curve experiment in the post processing window Once this is saved it is available for all DropQuant users Note using older standard curves in new experiments may lead to inaccuracies in case the old reference samples are not representative for the newer unknown samples It is therefore recommended to add references in the experiment to check to validity of the standard curve 67 The typical process of measuring samples using the standard curves method where the reference samples are part of the experiment is The user defines the standard curve method in the experiment definition window The user defines reference samples with reference concentrat
49. med C Please refer to the glossary for details on OD Sample size requirements and e recommended sample quantity External Characteristics The front panel of the instrument contains the centrally positioned microwell plate drawer to load and unload the DropFrame or the DropPlates and three indicator lights Green Orange and Red light Microplate drawer Indicator LEDs Front 1 The indicator LEDs show the status of the instrument a Green orange and red during the DropSense96 start up all three lights will flash simultaneously for about 10 seconds b Green and red DropSense96 is ready but no communication with a DropQuant running on the PC has been achieved check if DropQuant is running and if USB connection is OK c Green DropSense96 is ready and connected with the PC Green and orange light busy with a measurement Red error state Please refer to the troubleshooting section or contact your local distributor 2 Microplate drawer The microplate drawer moves in and outside of the instrument to load and unload the DropPlates A lid on the front panel closes the instrument automatically when the drawer is inside the instrument The drawer remains in the instrument during measurement cycle The drawer can accept different types of carriers as long as their outer dimensions are compatible with the SBS standard The plate carrier can be opened at any given time using the open tray button in the DropQua
50. nd perform experiments e DataViewer to analyze the experiments Computer requirements Computer requirements and software installation The operating software will only run on a PC meeting the criteria below No Mac versions of the software are currently available e Microsoft Windows XP Vista Windows 7 Windows 8 operating system e Microsoft NET Framework version 2 0 or higher e 1 5GHz or higher processor e 1GBor more of RAM 2GB if running Vista e 100 MB of free hard disk space for software installation e Free USB port the instrument can only be connected via the USB port e Microsoft Excel and Adobe pdf reader to manipulate archived data optional but essential for that Installation Updates Trinean s DropQuant Software installation and update works like many other windows software programs and both its installation and update should work by following the same procedure Login After inputting the user name and password a series of self tests will occur For more details on user names please consult the user name section in chapter 6 19 Home screen On this home screen it is possible to access the Measurement Database set up a New Experiment Measurement and to access the Info and Settings DropQuant Login Login Password connect to DropSense EXIT trinean DropQuant Log off and login Connect to the instrument SYSTEM INFO AND SETTINGS System info and Settings Open Tray ME
51. nd safety information in the user manual must be followed to ensure safe operation of the DropSense 96 and to maintain the DropSense 96 in a safe condition The following types of safety information appear throughout this user manual Details about these circumstances are given in a box the one below The advice given in this manual is intended to supplement not supersede the normal safety requirements prevailing in the user s country A A A gt Handling and positioning Always make sure the instrument is placed on a steady surface allowing easy access to its back It is extremely important that both the power button and the power cable at the back are always easily accessible Before moving repositioning the instrument always make sure that there s enough space for that Failing to provide adequate work space can cause material damage or injury to the handler Risk of personal injury When moving the instrument always make sure to use a 2 person lift as the apparatus weights more than 18kg Risk of personal injury and material damage W1 Improper use of the DropSense 96 may cause personal injuries or damage to the instrument The DropSense 96 must only be operated by appropriately trained and experienced personnel Servicing of the DropSense 96 must only be performed by Trinean trained service personnel or service technicians of an authorized agent Using the apparatus in a manner which is not specified in this user ma
52. nt 2010 10 08 16 11 2010 081 6 3mg ml 0 2mg 12 5mg m o amg mi 0 7 25mg m Lm 037 0 05mg 20 ay DESCRIPTION Purified Proteins BSA dilution series SOLVENT water based solvents incl TE PBS RT buffer MATERIAL _ eS Purified Proteins BSA 0 1mg mi 0 2mg mi DROPPLATES 1 Droprtet a a i somg m 0 0 8mg mi 1 6mg ml 1 55 mg ml 12 5mg mi BACK TO DATABASE EXPORT 43 Available columns Selected columns Exporting data 5 On DropQuant DataViewer exporting couldn t be easier Almost on every screen it is possible to export the measured data either to excel or one of the many other file types we have available or into a LIMS system Export Results When exporting results it is possible in advance to select which information to export Export Results Export Outliers Export Normalization trinean Select elements to Export Export Results Export Outliers Export Normalization All available columns Selected columns Export as Median A260 10mm DropPlate ID Include Summary Avg A260 10mm Pump Include 96 well plate layout StDev A260 10mm A260 A280 CV A260 10mm Materia direct print Export A280 10mm A260 10mm dt Raw A340 10mm Concentration mg ml settings Material DropPlate ID A260 A280 Material A260 Concentration Date 10mm krer DropPlate 1 ial am basal 0 03 Commer o o0 en az j o a oo a e DropPlate1 o 060 237 O T S O O E E S
53. nt software Do not obstruct the movement of the tray When reading is complete the drawer of the DropSense96 opens allowing customers to remove the DropPlate The drawer closes automatically after 30 seconds this feature is not active when the DropSense96 is integrated 17 erfean DropSense 96 Pewee Polychromatic microplate reader nascent B Art Nbr 101096 S N 2012 24 VDC B 9050 Gentbrugge a Belgium Made in Europe Crinean www trinean com j ha Back 2 3 Power switch Power cord receptacle plug the power cord in here USB port Label with line voltage rating cautionary information and serial number It is required to use the Trinean USB cable including the ferrite filters No stable operation of the instrument can be guaranteed if a different USB cable is used for example if a longer cable or if a cable without the filtering elements is used AUNE The power supply The DropSense instrument uses an external AC DC power supply similar to a laptop power supply which is included in the shipment The power supply input is compatible with the US standards 110VAC and European standards 230VAC Depending on the customers location a US type power cord or EU type power cord is included with the instrument This external power supply delivers 24VDC to the instrument 18 Software 2 DropQuant Trinean s DropQuant Software is divided in 2 important packages e DropQuant to define a
54. nual can impair the protection provided by the equipment Risk of personal injury and material damage W2 Do not attempt to move the DropSense 96 during operation Explosive atmosphere W3 The DropSense 96 is not designed for use in an explosive atmosphere Damage to the instrument C1 Direct sunlight may bleach parts of the instrument and cause damage to plastic parts The DropSense 96 must be located out of direct sunlight A A Electrical safety Note Disconnect the line power outlet before servicing A A To ensure satisfactory and safe operation of the DropSense 96 follow the guidelines below The line power cord must be connected to a line power outlet that has a protective conductor earth ground If the instrument becomes electrically unsafe prevent other personnel from operating it and contact your local distributor or Trinean Service support The instrument may be electrically unsafe when Damage to the instrument C2 Avoid spilling water or chemicals onto the DropSense 96 Damage caused by water or chemical spillage will void your warranty In case of emergency switch off the DropSense 96 at the power switch and unplug the power supply from the power outlet Moveable parts While operating the instrument please beware of any moveable parts Electrical hazard W4 Any interruption of the protective conductor earth ground lead inside or outside the instrument or disconnection of the protective conduct
55. or terminal is likely to make the instrument dangerous Intentional interruption is prohibited Lethal voltages inside the instrument When the instrument is connected to line power terminals may be live Opening covers or removing parts is likely to expose live parts Electrical hazard W4 Only use the provided power supply and supply cord For replacements contact your local distributor Use of wrong power supply may cause fire due to overheating The line power cord appears to be damaged It has been stored for a prolonged period of time in conditions which are outside of the normal storage conditions It has been subjected to severe transport stresses Biological safety Samples Samples may contain infectious agents You should be aware of the health hazard presented by such agents and should use store and dispose of such samples according to the required safety regulations Samples containing infectious agents W5 Some samples used with this instrument may contain infectious agents DropSense96 is not intended to be used for biological samples or infectious agents within the WHO risk class 3 and 4 Handle such samples with the greatest of care and in accordance with the required safety regulations Always wear safety glasses 2 pairs of gloves and a lab coat The responsible body e g laboratory manager must take the necessary precautions to ensure that the surrounding workplace is safe and that the instrument operat
56. ore detailed view that it s not possible with other tabs a Results Results trinean REPORT DISPLAY Select InputWell 0 05mg ml 3 1mg ml 0 1mg ml 6 3mg ml 0 2mg ml aput Well Selection 0 4mg ml 25mg ml PBS 1 6mg ml 0 05mg ml 3 1mg ml 0 1mg ml 6 3mg ml LID LIE LI LIS aS E E 10mm Absorbance E et ee ee Plate verview a canon an aw OC OC ACC IL o goon COOC CACCE mm mm a LIL BACK TO DATABASE SINGLE WELL DISPLAY autoscale COLUMN DISPLAY show cursor OD 10mm 280 300 320 340 Wavelength QUICK EXPORT TO XLS EXPORT Export Button subtract Concentration mg ml Concentration F1 11 87 mg ml Position F1 Type sample Blank average of blanks Name 12 5mg ml Concentration 11 87 mg ml A260 10mm 4 65 A280 10mm 7 91 Raw A340 10mm 0 15 A260 A280 0 59 blank Blank background correction linear EEE om W single point Background Correction At the top the customer can select the show cursor option This will show a cursor on the spectrum view window allowing readout of the OD at any wavelength SINGLE WELL DISPLAY OD 10mm autoscale 260 280 300 320 340 Wavelength COLUMN DISPLAY hide cursor 96 WELL DISPLAY wavelength nm 10mm Absorbance 3 30 A260 Concentration ng ul F1 164 48 ng ul Position F1 Type sample
57. orrect and more accurate measurements it is essential that a minimal amount of sample is dispensed for correct filling of the measurement chambers allowing precise measurements Although this volume range takes a pipetting error into account it is best to use a precision pipette e g 0 5 5 ul with precision tips to ensure that the recommended sample quantity is dispensed The DropPlate S The DropPlate S looks very similar to the DropPlate D however the meander reservoir is shorter and due to having only one measurement chamber its maximum range is lower than on the D The path length of the cuvette is 0 5mm and with a 0 1 40 OD range for a 10mm equivalent path the DropPlate S is ideally suited for quantification of biomolecule samples derived from automated extraction procedures with a stable yield For example extracted human genomic DNA tends to be within a 50 1000 ng ul concentration range and it can be quantified using the DropPlate S Since the DropPlate S contains only one micro cuvette instead 15 of two like the DropPlate D the measuring time for a full 96well DropPlate S is decreased by half to about 4 minutes DropPlate S Input well Storage reservoir Micro cuvette Measurement cycle EE i Step 1 Step 2 Sample is dispensed into Sample is drawn into the the input well manually or by reservoir by capillary action 2 liquid dispensing robot for siorage and sample we protection absorption measurement is peror
58. ors are not exposed to hazardous levels of infectious agents as defined in the applicable Safety Data Sheets SDSs or OSHA ACGIH t or COSHH documents Venting for fumes and disposal of wastes must be in accordance with all national state and local health and safety regulations and laws OSHA Occupational Safety and Health Administration United States of America t ACGIH American Conference of Government Industrial Hygienists United States of America COSHH Control of Substances Hazardous to Health United Kingdom Chemical safety Toxic fumes If working with volatile solvents or toxic substances you must provide an efficient laboratory ventilation system to remove vapors that may be produced Waste disposal In case plasticware contain hazardous chemicals or contagious biohazardous materials Such wastes must be collected and disposed of properly according to local safety regulations Mechanical To ensure satisfactory and safe operation of the DropSense 96 follow these guidelines e Use only recommended consumables slides e When powering down the instrument or when powering it up please do not stand or support your weight on the instrument Rotate the instrument or try to access it from behind by rotating the instrument Maintenance safety Perform the maintenance as described in Annex B Your local distributor and or Trinean charges for repairs that are required due to incorrect maintenance Risk of personal inj
59. ow 1 In DropQuant go to System Info and Settings and press the Wavelength calibration button Settings trinean system Info and Settings System Info Operating System windows 7 Professional DropQuant version jis Computer PDFCreator version SQLite installed Teamviewer version Application data permissions ok Disk Total diskspace MB Free diskspace MB DropSense DropSense Serial Number none L151 Internal bar code reader Firmware version Last Calibration 24 11 2014 2011 109 171 fTrinean Support v5 exe CO h fz ooo Nr of experiments 1931 Nr of measured plates fiass Nr of measured wells feses Nr Pulses Lamp Jazza o Stray light correction fversion 2 Nr Pumpcycles pump0 17537 Instrument Description FT245R USB FIFO Nr Pumpcycles pump1 117284 Electronics Serial Number a900ernIw Electronics ID 1432940281 OPEN TRAY Pump Time s pump s0701 Purnp Time s pump1 79520 Move to safe position Status 1Q0Q Edit Settings Diagnostics Potassium dichromate test Quick system check Check bar code reader Generate Status Zip File Create IQOQ report f Calibration administrator Wavelength calibration 2 Follow the onscreen instructions Performing wavelength calibration 5 Please place the optical calibration plate onthe tray inthe orientation as shown inthe picture and press OK Cancel
60. pQuant DataViewer starts allowing the user to access options designed to help interpret the measurements performed with the DropSense 96 Starting DropQuant Dataviewer rlease Walt When opening an experiment 4 tabs are visible to all users Report Display Single Well Display Column Display 96 Well Display a trinean Results REPORT DISPLAY SINGLE WELL DISPLAY COLUMN DISPLAY 96 WELL DISPLAY OD 10mm Select column al ii 39 This window allows the user to have a classical overview of the measured samples a Results z trinean Results REPORT DISPLAY SINGLE WELL DISPLAY COLUMN DISPLAY 96 WELL DISPLAY Select column OD 10mm 11 BACK TO DATABASE ii Similar to the 96 Well display but with a column view instead Results X9 trinean Results REPORT DISPLAY l SINGLE WELL DISPLAY COLUMN DISPLAY OD 10mm Concentration mg ml 23 20 15 10 5 2 3 2 2 230 250 275 300 325 350 375 400 230 250 275 300 325 350 375 400 230 250 275 300 325 350 375 400 230 250 275 300 325 350 375 400 coment Seam emg Sam 23 23 23 23 15 15 15 10 10 10 5 5 pm 250 275 300 325 350 375 400 ain 250 275 300 325 350 375 400 250 275 300 325 350 375 400 game 250 275 300 325 350 375 ne ma ee BACK TO DATABASE EXPORT 40 Probably the most interesting view on the results window as it allows the user to change parameters in order to analyze the results It also allows the user to have a m
61. ple for example the 260 280 and 260 230 ratios for DNA samples etc 11 K cDrop builds further on this selection 2 General UV vis mode The result of the measurement is the full absorption spectrum as a function of wavelength range 230nm 750nm 3 Standard curves a basic standard curve method is available since DropQuant 1 2 The user defines the reference samples and their corresponding reference values and the unknown samples DropQuant constructs then the standard curve and calculates the concentration of the unknown samples based on that information 4 cDrop a software extension that performs mathematical analysis on the UV Vis spectra to extract information on the content of the samples in contrast to the direct quantification method that uses the absorption information at a single wavelength to calculate the concentration of the sample cDrop is a software add on It is sold as a separate product but it integrates perfectly on the DropQuant software The small sample requirement using the DropPlates makes the DropSense96 the perfect solution for many applications e Measuring the concentration A260 nm and purity of nucleic acid samples DNA RNA oligos e Fluorescent dye labeling density of nucleic acid microarray samples e Purified protein analysis A280 nm e Quantification of fluorescent dye labeled proteins e API quantification Active Pharmaceutical Ingredient e General UV Vis spectrophotometry Z Trinean off
62. rformance test For more information regarding system requirements and purchase details please contact sales support at sales trinean com 7 What software add ons are dongle protected The list of modules is e cDrop e DropControl Service 8 Can Iuse my dongle after an upgrade of the software add on program You can upgrade the software add on program to newer versions and still use the old dongle and activation code you will have access to all functions that are allowed by the dongle 9 What if lose the activation code You can always ask Trinean a copy of the activation code 10 What if lose the dongle The dongle is unique there are no copies available This means you have to buy a new dongle 11 Can I copy the dongle No 12 changed measurement database but cannot see the measurements Check if the measurements are done by same user account If measurements in database are measured by user different from the user currently logged in the DropQuant software then they will not be visible Change to the administrator account to see all measurements defined in the user database of that DropQuant installation Measurements done by service are never visible by lab manager or standard users These measurements are visible by service level only This may happen when doing service interventions where the measurements are done under username service thus not visible to other users 13 cannot open import a DropFile D
63. ropQuant and cDrop cannot import zipped experiments The software can only import the experiment directory typically a directory with name experiment_1234 Most probably the version of the DropFile is more recent than the version of the DropQuant software You can check the version of the installed DropQuant from the settings and tools login as administrator You cannot check the version of the DropFile It is also possible to look up the DropQuant version that was used during the measurements You can find this by opening the log file which is part of the experiment directory 63 mar aPT MED 012536 Que O S Down mem B C Documents and Settings ronny bockstaele Bureaublad cDrop demo measurements Experiment _0 12536 _1 DropPlate 1 cdrop Configuration HA CDROP bestand 3 808 kB 1_DropPlate 1 S Analysis_1_DropPlate 1 cDrop pot e HA CDROP bestand 6 123 kB P log0 Kladblok 5 670 kB f Bestand Bewerken Opmaak Beeld Help saian aii details E ca naar _1_DropPlate 1 drop Bestands en maptaken DROP bestand Een nieuwe map maken 8 Deze map op het web publiceren G2 Deze map delen logO Tekstdocument 67 kB Andere locaties Drop demo measurements H Mijn documenten 4g Deze computer Q Mijn netwerklocaties a opQuant version version 11 0 22 Instrument ID 2009 025 Details Experiment_012536 Bestandsmap skip lines
64. rrection can be selected as for other nucleic acid samples The sequence can be inputted from the sample definition screen using the Enter check sequences button 1 First define the samples including the blank source plate definition and other information After the samples are defined the sequence information can be defined 2 Click the Enter check sequences button and a table with the wells defined on the plate will appear 28 Overview Oligo Sequences Select disposable DropPlate 1 y Select input method Add p pp or ppp to the 5 or 3 end for quantification of mono di or triphosphorylated oligonucleotides Oe For DNA enter a sequence with A T C G R Y M K S W H B V D N for RNA enter a sequence with AUCG E amp MW Position Sample Name Sequence DNA RNA E MW Conc Factor amp F as EXPORT XLS Available we have 2 different input methods Sequence Type in the oligonucleotide sequence or copy paste them from an excel table The software will calculate the properties such as the extinction coefficient the molecular weight and the concentration factor All the information can be saved in excel by pressing the export excel button at the bottom of the screen Phosphorylation at both the 5 and 3 end can also be included by adding the right number or the letter p to the sequence Press Save to return to the Pla
65. s as samples Clear this plate Export Import Plate Layout In order to speed up the process of starting a measurement it is possible to pre define a plate layout Several formats are accepted such as txt csv and xls amp trinean Plate Layout DropPlate 1 Ba Be 2 ee ee Ee ee ee ee ee A Water _ dsDNA_100ug dsDNA_50ug dsDNA SOug B __ dsDNA_100ug dsDNA_100ug dsDNA_50ug dsDNA _50ug BS aa aa aa O O D dsDNA_100ug dsDNA_100ug dsDNA_50ug dsDNA_50ug E __ dsDNA_100ug dsDNA_100ug dsDNA_50ug dsDNA _50ug F __ dsDNA_100ug dsDNA_100ug dsDNA_50ug dsDNA_50ug G _ dsDNA_100ug dsDNA_100ug dsDNA_50ug dsDNA_50ug H__ dsDNA_100ug dsDNA 100ug dsDNA_50ug Water Import Import of plate layout An import function Import Plate layout is foreseen at the bottom of the Plate Layout screen for fast and easy entering of plate layout information Several types of files excel txt and csv can be imported and the data can be reformatted into a dedicated DropQuant software table for quick plate layout configuration 24 Selected La template preview Import format settings Imported data preview File type Open xls selection template Import Samples trinean Import Samples f xis xlsx csv y Import format Default converted sample s 0 more info selected sample s 0 selectall deselect all Source Source DropPlate DropPlate S
66. se the DropQuant will check the DropPlate type and expiry date during the measurement not before the measurement starts In that case it is the responsibility of the customer to connect the results of the DropQuant measurement with the source plate position information It is assumed the customer has saved this information during the dispensing The internal barcode scanner is an option of the DropSense instrument Contact the service engineer to install the barcode scanner on the instrument Use of an external scanner for scanning the barcode for use in the plate definition It is possible to use part of the barcode functionality when using an external barcode scanner that acts as an external keyboard This offers next functionality 50 e The name can be scanned in a convenient way and the DropQuant program will check the type of disposable and expiry date of the DropPlate e The name can be scanned and the available wells database will be checked to disable the wells on that plate that have been used before K It is not possible to check the order of the plates inserted in the instrument when S doing an experiment with multiple DropPlates This check is available for internal barcode scanner only There are many types of external barcode scanners acting as a keyboard first click with mouse on the input field in the DropQuant software so that the cursor is at the good position Then scan the barcode the code will be inserted autom
67. surement Database location The experiments raw data is stored in a measurement database location This directory contains a subdirectory per experiment with all raw data The location of this directory can be changed by clicking the measurement database location button K Indexing of large databases can take a long time especially when the database location is on a network drive 53 Formats The formats are not common for all DropQuant users Each user defines his formats and these are by default not visible to other DropQuant users However the administrator can make one or more of his formats visible to all DropQuant users Edit format settings Please select which formats should be visible for all users If you uncheck a format it will only be visible for you Experiment Definition Format Prot_IgG Unlabeld Hyd y Default predefined Export Formats Default Nucleic Acids Available well database Administrator level users have access to the used well database management tools via the settings and tools window The user can deactivate this function and change its location It is recommended to select a network drive for this so that different PCs can have access to the same available wells database It is also possible to e Remove entries from the database e Cleanup the database e Search the database 54 Available wells database C ProgramData Trinean Used_Wells db Select new location
68. te layout screen E amp MW Here it is possible to enter individual or replicated extinction coefficients and molecular weight information When using the Replicates function entering the sequence of one sample will automatically leads to the fill in of the sequence and data for all its replicates C7 Please don t confuse Replicates with the F button available on the same screen The IN A last one is meant to be used on the Standard Curves application Proteins Measuring multiple proteins with different extinction coefficients when measuring several proteins it is possible to set that information before the measurement 29 E BEREREEa OOOGOEE8 OOOOOO00 gt DOOEOOOe 3 DOOOOOEO gt prrr0r00 O00000 OOOOOUOOO 0000000 OOOOUOOU DOOOOUoooO Overview E1 Select disposable DropPlate 1 v Position Sample Name 1 30 General UV Vis and Standard Curve In addition to the other sample type options it is also possible to perform a General UV Vis measurement or a Standard Curve Standard curves For more information on standard curves please read the extra information on the Annex 1 Theory Defining standard curve experiment As explained in the introduction standard curves can be defined inside the experiment or by selecting a standard curve from an earlier experiment Both methods are explained in this paragraph Case 1 the standard reference samples are p
69. ting 61 Solving an issue 61 FAQ list 61 Annex A Theory 65 DNA RNA concentration and purity determination 65 Protein concentration and purity determination 66 General UV Vis spectrophotometry 67 Principle of standard curve measurements 67 Annex B Maintenance 69 Cleaning the DropSense 96 69 Maintenance 69 Annex X Glossary 70 This manual is published by TRINEAN NV SA Questions or comments regarding the content of this manual can be directed to the address below or to your TRINEAN representative TRINEAN NV SA Dulle Grietlaan 17 3 B 9050 Gentbrugge Belgium Tel 0032 9 2727535 Fax 0032 9 2727539 info trinean com www trinean com DropSense 96 and DropPlate16 96 are trademarks of TRINEAN NV SA This document is the copyright of TRINEAN and must not be copied or reproduced in any form without prior consent TRINEAN reserves the right to make technical improvements to this equipment and documentation without prior notice as part of a continuous program of product development This manual supersedes all previous editions Trinean warranty statement Warranty and Special Provisions Limited Warranty TRINEAN products are fully guaranteed for one year against defects in parts materials and workmanship Defective parts and materials will be replaced or at the discretion of TRINEAN repaired at no charge for a period of one year provided that the products are utilized and maintained in accordance with the instructions in
70. uant DataViewer ID Date User Name Description Change user ZIP selected experiment Delete selected experiment Import Database REFRESH NEW SEARCH OPEN DATA Change user Set a different user for the selected measurements K This is available for all measurements owned by user levels guest user and S administrator If for example the measurements are done by a service engineer then the administrator is not able to change the owner of these measurements Delete selected experiment Deletes the raw data of the experiments on the disk and removes the entries from the database This completely removes the experiments There is no way to recover the data afterwards ZIP selected experiment Zips a selected experiment to a desired location Import database This function imports a set of experiments from a folder The experiments must be copied to a temporary directory and unzipped in case these new experiments are zipped 56 Diagnostic and calibration tools Optical calibration 3 With the new DropQuant v1 5 it is possible for the user to perform an optical calibration of the DropSense96 For this calibration an optical calibration plate is required This calibration plate can be purchased from your local distributor This calibration should also be performed depending on the instrument s usage at least once a year In order to do the wavelength optical calibration please follow the instructions bel
71. ucibility Although this volume range takes a pipetting error into account it is best to use a precision pipette e g 0 5 5 ul with precision tips to ensure that the recommended sample quantity is dispensed OD optical absorption measured by the instrument The OD10mm is the OD calculated as if the path length is 10mm the path length of a standard cuvette Recommended sample quantity volume to be dispensed in the input well to guarantee good and high yield measurements with the given measurement range and sample residence time Check the accuracy of the pipette and the tips to avoid the dispensed sample volume exceeds the maximal sample quantity 70
72. ury and material damage W6 Only perform maintenance that is specifically described in this user manual Risk of electric shock W7 Do not open any panels on the DropSense 96 Damage to the instrument C3 Do not use solvents or reagents containing acids alkalis or abrasives to clean the DropSense 96 Specs for DS96 Technical data Trinean reserves the right to change specifications at any time Environmental conditions Operating conditions Power Fuse Overvoltage category Data output Air temperature Relative humidity Altitude Place of operation 100 240 V AC 50 60 Hz Mains supply voltage fluctuations are not to exceed 10 of the nominal supply voltages External 24 V power supply designed for wide range of input voltages maximum power consumption approximately 15 W TIA OVC II USB 15 C t0 35 C 59 F 16 95 F Max 75 noncondensing Up to 2000 m 6500 ft above mean sea level MSL For indoor use only Transportation conditions Air temperature Relative humidity Storage conditions Air temperature Relative humidity 252C to 602C 132F to 1402F in manufacturer s package Max 75 noncondensing 52C to 402C 412F to 1042F in manufacturer s package Max 85 noncondensing 10 DropSense96 at a glance DropSense96 Overview The DropSense96 platform is designed for measuring the UV Vis absorption spectrum of microliter sized samples suitable for both manual and automated workf

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