YSA-400 User Manual
Contents
1. 31 5 5 Cell Cultures The YSA 400 micro spectrophotometer can measure the concentration of suspended cell cultures using one path length 1mm 5 5 1 Sample size requirements Recommended volume 2 0uL 5 5 2 Measurement range 0 1Abs 75Abs Typical reproducibility SD Abs CV Sample range 0 1Abs 5SAbs 0 1 Sample range 5 75Abs 2 5 5 3 Work interface Click the Cell Cultures button in the measurement type select area and enter the cell cultures measurement interface Wavelength and absorbance The blue vertical measurement line corresponds to the wave length value of Xc Pos 600 and the absorbance corresponds to the Xc Value i e the value of Xc Value is the absorbance at 600nm path length 10mm The user can move the vertical measurement line to the appropriate place to get the corresponding absorbance Path length The software provided one path length 1mm for measuring cell cultures A scan curve will appear in the graph display area when measuring a sample 5 5 4 Measurement procedures For detailed measurement procedures please see 4 2 1 At first measure a blank using the culture solution without cells by dropping it onto the lower measurement pedestal and clicking Blank button to measure and store the blank 32 2 Wipe the solution from the measurement pedestals with clear absorbent paper 3 Then measure the samples 4 The result values and graph of the sample will app2. BCA Bradford and Lowry For example select BCA for measuring the proteins with BCA protein assay kit pr BoSU0 BEacrodpect coPhotolecter Filet View Tool Tl Help iH Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample Standard Sample Test Type f F Sample Name Sample Sample ID D 10mm Absorbance Blank Save Graphic Show Report View History Data 610 0 650 0 690 0 EA Wave Length nm Xa Pos Xb Pos Xc Pos Xa Value Xb Value Ac Value 28 2 Click Standard to measure the standard proteins 3 At first measure a blank 4 Wipe the solvent from the measurement pedestals 5 Measure at least five standard proteins with different concentrations 6 Click the right mouse button in the standard table and select the Work Curve to generate a standard curve 7 Then click Sample to measure the samples 8 Measure samples and the result values and graph will appear in the work interface in seconds 8 Click the Save Graphic button after measured a sample if you want to store the graph 9 Click the Show Report button after the samples measured to output the result by Excel 29 5 4 UV VIS The YSA 400 micro spectrophotometer can make UV VIS full spectrum 200 850nm absorbance scan 5 4 1 Sample size requirements Recommended volume 2 0uL 5 4 2 Measurement range 0 1Abs 75Abs Typical reprodu
3. Teel Ti Help H Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample amp Test Type Sample Name Sample Sample ID a c 3 a E E Save Graphic Show Report View History Data 259 0 272 0 2080 338 0 311 0 Wave Lengthinm a Pos Xb Pos xc Pos 260 280 Xa Value Xb Value xc Value ng ul 6 Click the Save Graphic button after measured a sample if you want to save the graph 7 When all the double stranded DNA samples measurement finished click the Show Report button to output the results by Excel 22 5 2 Protein A280 The YSA 400 micro spectrophotometer can measure the concentration of proteins with two methods protein A280 and the protein assay kits Because proteins have the highest absorption peak at 280nm the concentration of proteins can be calculated and reported by the software based on the Beer Lambert law 5 2 1 Sample size requirements Recommended volume 2 0uL 5 2 2 Measurement range BSA 0 1 100mg mL Typical reproducibility SD mg mL CV Sample range 0 1 10mg mL 2mg mL Sample range gt 10mg mL 2 5 2 3 Work interface Click the Protein button in the measurement type select area and enter the protein measurement interface as below pnd K5500 MicroSpectroPhotoMeter File F View Tool T Help H Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample amp Test Type Sample Name Sample Sa
4. 4 Functions of work interface Open the YSA 400 software and enter the work interface of the YSA 400 micro spectrophotometer as below pn K5500 MicroSpectroPhotoMeter File F View Tool T Help H Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample amp Test Type Sample Name Sample Sample ID 0 10mm Absorbance Blank Save Graphic Show Report 272 0 285 0 d 260 230 Wave Length nm NAN Xa Pos Xb Pos Xc Pos 260 280 Xa Value Xb Value Xc Value ng ul For easy introduction the work interface can be divided into five parts menu bar measurement type select area graph display area data display area and measurement function area 4 1 Menu bar There are four main menus in the menu bar File View Tool and Help ne a ee ee File F VYiew Tool fT Help H ee File An Exit menu is under the File and the program will exit if click the Exit File View Tool CI Help H Exit Ctrlti nen dd Protein View There are four submenus under the View Auto Scale Multi Draw Show Grid and Color Solution File F view Tool Tl Help H Auto Scale Ctrlta Multi Draw Ctrlti of Show Grid Ctrl G i Color Configure CtrltC Auto Scale submenu enables the user to scale the Y axis The default setting Nucl can automatically scale the Y axis based on the sample The
5. Micro Volume UV Spectrophotometer Model No YSA 400 User Manual Version V1 00 Software Version V1 00 Hardware Version V1 00 Yu Shing Biotech Ltd Declaration Yu Shing Biotech Ltd has the copyright of this manual which is a guide for using YSA 400 micro spectrophotometer You can t copy modify or post it to the Internet without authority granted by Yu Shing Biotech Ltd And it should be read carefully when you install configure and use YSA 400 micro spectrophotometer We will have no responsibility for the loss of date casualties or other loses induced by incorrect operation or other accidents which are mentioned or not in the manual Now enjoy the convenience of YSA 400 micro spectrophotometer if you obey the rules Package List Every product has following objects They should be checked carefully when you open the package for the first time Yu Shing Biotech Ltd and the dealers will replace relative articles for free if they are found different from the ones in the following table YSA 400 micro spectrophotometer 200 850nm The CD for YSA 400 nn 200 850nm USB connector 12V Power adapter Table of Contents 1 Introduction 2 Installation 2 1 Port connection 2 2 System requirements 2 3 Installing the software 2 4 Installing the drivers 3 General operation 3 1 Sample size requirements 3 2 Measurement procedures 3 2 1 Start 3 2 2 Blank 3 2 3 Measurement 3 2 4 Output 4 Functions of work
6. Show Report and View History Data as below Blank Save Grph Show Report View History Data The Blank button is used to measure and store the blank which must be measured before measuring the samples The Measure button is used to measure the samples after measured a blank and it is inactive and can t be used before a blank measured The Save Graphic button is used to save the graph after measured a sample The Show Report button is used to output the result data of the samples by excel View History Data enables the user to browse all the results measured before In addition there are Sample Name bar and Sample ID bar under the Sample amp Test Type The user can input a name in the Sample Name bar and a number in the Sample ID bar for the sample when measuring it 5 Measurement 5 1 Nucleic acid The YSA 400 micro spectrophotometer can measure the concentration and purity of nucleic acids because nucleic acids have the highest absorption peak at 260nm and the concentration of nucleic acids can be calculated and reported by the software based on the Beer Lambert law and the ratio of absorbance at 260nm and 280nm and the ratio of absorbance at 260nm and 230nm can assess the purity of nucleic acids 5 1 1 Sample size requirements Recommended volume 2 0uL 5 1 2 Measurement range DS DNA 2 3700ng uL SS DNA 2 2400ng uL RNA 2 3000ng uL Typical
7. Type When the user selects one of the BCA Bradford and Lowry Click the Standard button a table will appear under it as below Sample Standard The data will be shown in the table when measuring a standard protein The column of Ave Abs shows the absorbances of standard proteins And the user can input the concentration of the standard protein in the column of Conc A work curve will form by clicking the right mouse button in the standard table and selecting the Work Curve after measured all the standard proteins Delete All Item enables the user to delete all the data Delete Item can delete one row Load Standard File can load the data of a standard file saved in the computer Save Standard File can save the data of the standard protein 27 Sample Standard Delete All Item Ctrl I Delete Item Ctrl I fork Curve Ctrlti Load Standard File Ctrl L Save Standard File Ctrlts FF Wavelength and absorbance The vertical measurement line corresponds to the wavelength of 560nm for measuring the protein with BCA method 595nm for Bradford and 660nm for Lowry Concentration The concentration value of proteins will appear in the mg mL bar after measuring a sample each time and the unit is mg mL 5 3 4 Measurement procedures For detailed measurement procedures please see 4 2 1 Click the Sample amp Test Type to select the right measurement type from
8. drop sample onto the lower measurement pedestal and click the measure button to measure the sample 7 The result values and graph of the sample will appear in the work interface in seconds 36 8 Click the Save Graphic button after measured a sample if you want to store the graph 9 Click the Show Report button after the samples measured to output the result by Excel 37
9. reproducibility SD ng uL CV Sample range 2 100ng uL 2ng uL Sample range gt 100ng uL 2 5 1 3 Work interface Open the YSA 400 software and the default interface is the nucleic acids measurement interface as below pn K5500 MicroSpectroPhotoMeter File F YView Tool T Help H Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample amp Test Type Sample Name Sample Sample ID 0 Blank Save Graphic Show Report 350 0 260 230 NAN 272 0 285 0 Wave Length nm Xa Pos Xb Pos Xc Pos 260 280 Xa Value Xb Value Xc Value ng ul Measurement type Click Sample amp Test Type to pop out a drop down list box including DS DNA 50 SS DNA 33 and RNA 44 for measuring double stranded DNA single stranded DNA and RNA respectively Sample amp Test Type Wavelength and absorbance There are three vertical measurement lines with different color pink yellow and blue in the graph of which the corresponding wave lengths are the values of Xa Pos 230 Xb Pos 260 and Xc Pos 280 in the data display area respectively and the corresponding absorbances are the values of Xa Value Xb Value and Xc Value respectively i e the values of Xa Value Xb Value and Xc Value are the absorbances at 230nm 260nm and 280nm respectively path length 10mm 260 280 The ratio of absorbance at 260nm and 280nm can assess the purity of nucleic acids Generally the ratio
10. Report View History Data Dyei Cy3 460 0 25 0 686 0 720 0 650 0 z Wave Length nm Abs 0 000 pmotl ul 0 000 Xa Pos Xb Pos Xc Pos Dye2 Cy5 E oo poo 9s0290 Xa Value Xb Value Ac Value nglul 2 s Measurement type Nucleic Acids Click Sample amp Test Type to pop out a drop down list box including DS DNA SS DNA and RNA for measuring double stranded DNA single stranded DNA and RNA respectively at 260nm Fluorescent dyes 10 fluorescent dyes are provided by the software The respective extinction coefficient absorbance wavelength and 260nm and 280nm corrections are shown in the table by clicking the Dye Editer submenu under the Tool main menu Dye Editer Dye Chromophore Editor 0 040000 ma p pe pe 150000 000000 0 050000 Cy5 250000 000000 650 0 000000 0 050000 Alexa Fluor 468 10000 000000 495 0 300000 0 110000 Alexa Fluor 546 104000 000000 556 0 210000 0 120000 Alexa Fluor 555 150000 000000 555 0 040000 0 080000 Alexa Fluor 594 73000 000000 590 0 430000 0 560000 Alexa Fluor 647 239000 000000 650 0 000000 0 030000 Alexa Fluor 660 132000 000000 663 0 000000 0 100000 Cy3 5 150000 000000 581 0 080000 0 240000 Cy5 5 250000 000000 675 0 050000 0 180000 Save amp Exit Exit The user can also select the dyes from the drop down list box The default setting is that the Dye1 is Cy3 and the Dye2 is Cyd Wavelength and absorbance The pink vert
11. amples or after reselecting a measurement type each time 3 Then open the sampling arm and drop 0 5 2uL solvent onto the lower measurement pedestal 10 4 Close the sampling arm and click the Blank button The sample column is automatically formed as below and the software will automatically measure and store the result of the blank 5 Open the sampling arm and wipe the blank solvent from the measurement pedestals with clear absorbent paper 3 2 3 Measurement 1 Open the sampling arm and drop 0 5 2uL sample onto the lower measurement pedestal 2 Close the sampling arm and click the Measure button and the software will automatically measure the sample 3 The result values and graph of the sample will appear in the work interface in seconds If you arent sure about the result or you don t think the curve is normal the reasons may be that the sample column isn t formed or the blank doesn t work well Please check whether the sample column is formed and measure it again or measure the blank again and then measure the sample again 4 Click the Save Graphic button after measured a sample if you want to save the graph 5 Open the sampling arm and wipe the sample from the measurement pedestals with clear absorbent paper after each time a sample measured 3 2 4 Output Click the Show Report button after the samples measured to output the result data of the samples by excel
12. cibility SD Abs CV Sample range 0 1Abs 5Abs 0 1 Sample range 5 75Abs 2 5 4 3 Work interface Click the UV VIS button in the measurement type select area and enter the UV VIS full spectrum absorbance scan interface Wavelength and absorbance There are three vertical measurement lines with different color purple yellow and blue in the graph of which the corresponding wave lengths are the values of Xa Pos Xb Pos and Xc Pos in the data display area respectively and the corresponding absorbances are the values of Xa Value Xb Value and Xc Value respectively User can move the measurement lines to the appropriate place based on the sample 5 4 4 Measurement procedures For detailed measurement procedures please see 4 2 1 At first measure a blank using the solvent by dropping it onto the lower measurement pedestal and clicking Blank button to measure and store the blank 2 Wipe the solvent from the measurement pedestals with clear absorbent paper 30 3 Then measure the samples 4 The result values and graph of the sample will appear in the work interface in seconds and the typical absorption spectrogram of K2Cr 07 is displayed as below Bute eet Firetetnh UW WIS Celi Cuitre Mcredstey 5 Click the Save Graphic button after measured a sample if you want to store the graph 6 Click the Show Report button after the samples measured to output the result by Excel
13. ear in the work interface in seconds and the typical absorption spectrogram of BCA is displayed 5 Click the Save Graphic button after measured a sample if you want to store the graph 6 Click the Show Report button after the samples measured to output the result by Excel 33 5 6 Micro Array The YSA 400 micro spectrophotometer can measure the concentration of fluorescent dye labeled nucleic acid micro array for it can measure the concentration of the nucleic acid and the dyes at the same time The concentration of nucleic acids and the fluorescent dyes can be calculated and reported by the software based on the Beer Lambert law The software provided 10 fluorescent dyes for you to select 5 6 1 Sample size requirements Recommended volume 2 0uL 5 4 2 Measurement range DNA 2 750ng uL Typical reproducibility SD ng uL CV Sample range 2 100ng uL 2ng uL Sample range gt 100ng uL 2 Cy3 0 2 100pmol uL Typical reproducibility SD ng uL CV Sample range 0 2 4 0pmol uL 0 2pmol uL Sample range gt 4 0pmol uL 2 5 6 3 Work interface Click the Micro Array button in the measurement type select area to enter the micro array measurement interface as below 34 BRO500 SicrospectroPhotoleter Pile View Tool Help Q Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample amp Test Type Sample Name Sample Sample ID o Blank Save Graphic r Show
14. ical measurement line corresponds to the wavelength of 260nm for measuring the nucleic acids and the corresponding absorbance are shown in the Xa Value The yellow and blue vertical measurement lines correspond to the wavelengths of the dyes for measuring the dyes and the corresponding absorbance are shown in the Dye1 Abs and Dye2 Abs respectively Concentration The value of nucleic concentration will appear in ng uL bar after measuring a sample each time and the unit is ng uL The value of dye concentration will appear in the uM bar and the unit is pmol uL 5 6 4 Measurement procedures For detailed measurement procedures please see 4 2 1 Click the Micro Array to enter the micro array measurement interface 2 Click the Sample amp Test Type and select the right measurement type of nucleic acid from DS DNA 507 SS DNA 33 and RNA 44 For example select SS DNA 33 for measuring the single stranded DNA and dye samples 3 Click the dyes button to select the right measurement type of dye from 10dyes For example choose Cy3 for measuring the SS DNA and Cy3 samples and or choose Cyd for measuring the SS DNA and Cy3 and or Cy5 samples 4 At first measure a blank using the solvent by dropping it onto the lower measurement pedestal and clicking Blank button to measure and store the blank 5 Wipe the solvent from the measurement pedestals with clear absorbent paper 6 Then
15. interface 4 1 Menu bar 4 2 Measurement type select area 4 3 Graph display area 4 4 Data display area 4 5 Measurement function area 5 Measurement 5 1 Nucleic acid 5 1 1 Sample size requirements 5 1 2 Measurement range 5 1 3 Work interface 5 1 4 Measurement procedures 5 2 Protein A280 5 2 1 Sample size requirements 5 2 2 Measurement range 5 2 3 Work interface 5 2 4 Measurement procedures 5 3 BCA Bradford and Lowry 5 3 1 Sample size requirements 5 3 2 Measurement range 5 3 3 Work interface 5 3 4 Measurement procedures 5 4 UV VIS 5 4 1 Sample size requirements 5 4 2 Measurement range 5 4 3 Work interface 5 4 4 Measurement procedures 5 5 Cell Cultures 5 5 1 Sample size requirements 5 5 2 Measurement range 5 5 3 Work interface 5 5 4 Measurement procedures 5 6 Micro Array 5 6 1 Sample size requirements 5 6 2 Measurement range 5 6 3 Work interface 5 6 4 Measurement procedures 1 Introduction The YSA 400 micro spectrophotometer a new type full spectrum 200 850nm spectrophotometer can measure 0 5 2 uL samples with high accuracy and reproducibility The samples can be measured by dropping them directly onto the lower measurement pedestal without absorption cell and also can be taken back after measuring if necessary This instrument don t need warm up after start and is operated simply and quickly with reporting directly the concentrations of samples The concentration range of samples measured by K5500 micro
16. is about 1 8 for relative pure DNA and about 2 0 for relative pure RNA A lower ratio indicates that the DNA or RNA may be contaminated by other things 20 260 230 The ratio of absorbance at 260nm and 230nm can also assess the purity of nucleic acids Generally the ratio is 1 8 2 0 for relative pure DNA and RNA The DNA and RNA may be contaminated by other things if the ratio is lower than 1 8 Concentration The concentration value of nucleic acids will appear in the ng uL bar after measured a sample each time and the unit is ng uL 5 1 4 Measurement procedures For detailed measurement procedures please see 4 2 1 Click the Sample amp Test Type to select the right measurement type from DS DNA 50 SS DNA 33 and RNA 44 For example select DS DNA 50 for measuring the double stranded DNA samples 2 At first measure a blank using the solvent by dropping it onto the lower measurement pedestal and clicking the Blank button to measure and store the blank 3 Wipe the solvent from the measurement pedestals with clear absorbent paper 4 Then drop sample onto the lower measurement pedestal and click the measure button to measure the sample 5 The result values and graph of the sample will appear in the work interface in seconds and the unique absorption spectrogram of DS DNA is displayed as below 21 de BOD BacroSspectroPhotolleter a i E File iF i aw
17. light will come on 4 Make sure the sampling arm is closed and the pedestals are clean or it will seriously impact the measurement result 5 Double click the K5500 exe icon on the desk or one click the K5500 exe icon from Start menu to start the K5500 software E5500 Install 6 Then the instrument starts self checking 7 If the following box pop up please press OK and check whether the instrument is connected correctly to the computer the sampling arm is closed and the pedestals are clean Then you must switch off the power of the instrument by pressing the switch and repeat 1 7 Please make sure the arn i down and the pedestals it clean Press OE 8 The above picture will disappear and the work interface will open as below when the self checking finished pnd K5500 MicroSpectroPhotoMeter File F YView Tool T Help H Nucleic Acid Protein UV VIS Cell Culture MicroArray Sample amp Test Type Sample Name Sample Sample ID 0 9 9 G F A 2 H E e Blank Save Graphic Show Report View History Data F TTE 4 260 230 Wave Length nm NAN Xa Pos Xb Pos Xc Pos 260 280 mm o NAN Xa Value Xb Value Xc Value ng ul Now you can measure the sample from here 3 2 2 Blank 1 You must choose the correct measurement type from the measurement type select area based on you samples before measuring 2 A blank must be measured before measuring the s
18. mple ID 0 10mm Absorbance Blank Save Graphic Show Report Seer 272 0 285 0 298 0 311 0 324 0 260 230 Wave Length nm Xa Pos Xb Pos Xc Pos 260 280 Xa Value Xb Value Xc Value mg ml Measurement type Click the Sample amp Test Type to pop out a drop down list box where A280 1 BSA 6 7 IgG 13 7 and Lysozyme 26 4 are designed for measuring protein at 280nm Sample amp Test Type A280 1 is designed for measuring the protein solution which produce an absorbance at 280nm of 1 0A path length 10mm at the concentration of 1 mg ml BSA 6 7 IgG 13 7 and Lysozyme 26 4 are designed for measuring the bovine serum albumin BSA immunoglobulin G lgG and Lysozyme respectively Wavelength and absorbance The blue vertical measurement line corresponds to the wave length value of Xc Pos 280 and the absorbance corresponds to the Xc Value i e the value of Xc Value is the absorbance at 280nm path length 10mm Concentration The concentration value of proteins will appear in the mg mL bar after measuring a sample each time and the unit is mg mL 5 2 4 Measurement procedures For detailed measurement procedures please see 4 2 1 Click the Sample amp Test Type to select the right measurement type from A280 1 BSA 6 7 IgG 13 7 and Lysozyme 26 4 For example select BSA 6 7 for measuring the bovine serum albumin sa
19. mples 2 At first measure a blank using the solvent by dropping it onto the lower measurement pedestal and clicking Blank button to measure and store the 24 blank 3 Wipe the solvent from the measurement pedestals with clear absorbent paper 4 Then drop sample onto the lower measurement pedestal and click the measure button to measure the sample 5 The result values and graph of the sample will appear in the work interface in seconds and the typical absorption spectrogram of BSA is displayed as below ET BacrospectroPhototleter i A Fila iF View Tooli Help la Nucleic Acid Protein UV VIS Cell Culture MicroArray Sam pleaTest Type Sample Name Sample Sample ID Measure Save Grphic Show Report Wiew History Data a720 285 0 298 0 311 0 EEES 737 0 Wave Length nm Ma Pos Xb Pos Ac Pos Ma Value Xb Value Mc Value mg ml 6 Click the Save Graphic button after measured a sample if you want to save the graph 7 When all the BSA samples measurement finished click the Show Report button to output the results by Excel 25 5 3 BCA Bradford and Lowry The YSA 400 micro spectrophotometer can measure the concentration of proteins of which the concentration is relatively lower or not pure using protein assay kits methods These methods include BCA Bradford and Lowry Please see the protein assay kits ins
20. new hardware and tip Find new hardware Windows will automatically open hardware installation guide Now select Install from a list or specific location Advanced as below aa ES hes We ee fe ee lt 5 ESSE EE RT Ocean Optics Us B2000 02 SR AES CD RBA HIRE aP MA ARTS EA TELF OJEE ee GES 5 1 O fie BPE He Se OEE sete FF ete lt b FEN lingi Click Next and enter driver options dialog box Click Don t search will choose the drivers to install as below SELE aT ee ee SWS BK Ocean Optics TS B2000 Si T N Rri Wait Windows system will automatically install the selected divers When the installation is finished a dialog box will appear as below r EG aT EE TORK ET PA Be AA oe ie elas se eRe Ocean Optics Us B2000 He DEAR Ri bite a E E FRR AH Click OK to finish the hardware installation Now Windows will tip you that the instrument is installed successfully and can be used 3 General operation 3 1 Sample size requirements Nucleic acids 0 5 2 0uL Proteins 0 5 2 0uL Lowry BCA and Bradford 0 5 2 0uL Cell suspensions 0 5 2 0uL Other solutions 0 5 2 0uL 3 2 Measurement procedures 3 2 1 Start 1 Connect correctly YSA 400 micro spectrophotometer to the computer 2 Connect correctly YSA 400 micro spectrophotometer to the power adapter 3 Switch on the power by pressing the switch then the red
21. ram designed by the software can automatically generate the standard curve after measured standard protein samples Then the concentration of the protein sample is report when the sample is measured UV VIS This type is designed for performing UV VIS full spectrum 200 850nm absorbance scan with two path lengths 1mm and 0 2mm imm 0 2mm Cell Culture This type is designed for measuring the concentration of suspended cell cultures with one path length 1mm Micro Array This is designed for measuring the concentration of nucleic acid at 260nm and the concentration of fluorescent dye at its highest absorption peak at the same time Software provided 9 florescent dyes 4 3 Graph display area The graph display area displays the scan curve of absorbance as below K E e ETIS LALL TETTETETT ELEELE se Bee ecb ELL LLL LLELLE ELLLER d o tty i EOE ls pA unc Ap nyc nym Y oy sgt ss gin cams a a npn cp ab say da npn cst cm npn ap a sn sp in yt in Tn cn ops in pt ee oe ee Oe Oe ee o m anaa as aaa aaa a T A ee One a a aa Gav aaa a Cee aa A eine 10mm Absorbance 0 35 220 0 233 0 246 0 259 0 272 0 285 0 298 0 311 0 324 0 337 0 350 Wave Length nm The red curve is the scan curve of absorbance generated each time after measured a sample There are three vertical measurement lines with different color pink yellow and blue in the graph of which the corresponding
22. rfect condition before leaving factory Clicking the UV or VIS will check the light intensity and produce curves of light intensity w W ai 00 1 I i aa I WA Laaa A MAJ ASIN am thor iM Ay Way W if yo val ry Uh N WA M Y i Dye Editor enables the user to save the information of fluorescent dyes from the table as below 14 Dye Editer Dye Chromophore Editor Alexa Fluor 468 Alexa Fluor 546 Alexa Fluor 555 Alexa Fluor 594 Alexa Fluor 647 Alexa Fluor 660 Cy3 5 Cy5 5 10000 000000 104000 000000 150000 000000 3000 000000 239000 000000 132000 000000 150000 000000 250000 000000 495 556 555 590 650 663 581 675 0 300000 0 210000 0 040000 0 430000 0 000000 0 000000 0 080000 0 050000 0 110000 0 120000 0 080000 0 560000 0 030000 0 100000 0 240000 0 180000 150000 000000 550 0 040000 0 050000 Cy5 250000 000000 650 0 000000 0 050000 __ et The respective extinction coefficient absorbance wavelength and 260nm and 280nm corrections are shown in the table Help The Help menu will help the user understand the majority of questions met during using the software There are About and Help submenus under the Help menu File l View Tool T Help OO Nucleic Acid Pr And clicking the Help submenu will open YSA 400 micro spectrophotometer user manual 4 2 Measurement type select area There are six items in the mea
23. s software requirements Microsoft Windows 2000 and XP or better operation system Microsoft Office 2000 or higher office software Adobe Reader 7 0 or higher version Hardware requirements CPU Intel Pentium IIl 800MHz or above EMS memory a minimum of 256MB Hard disk with a minimum of 50MB of free disk space 2 3 Installing the software Put the YSA 400 software CD into the CD drive Double click the CD catalogue open the Setup folder and run the YSA 400 Install exe file to start the installation process of the software Click Next choose installation catalog as below is E5500 HicroSpecrusPhotolleter Select Installation Folder The installer will install KS500 MicroSpecrumPhotoM eter to the following folder To install in this folder click Next To install to a different folder enter it below or click Browse Folder C Program FilesiKaiao BeijingiE5500 MicroSpecrumFhotoll Browse Disk Cost Install E5500 Micros pecrumPhotoM eter for yourself or for anyone who uses this computer f Evernone C Just me Cancel lt Back l Click Next and input the serial number Click Next to start the installation of the software Click Close when installation is finished The YSA 400 icon will automatically be formed on the desk CB4500 2 4 Installing the drivers After the instrument is connected correctly with the computer Windows will automatically scan the
24. spectrophotometer is 50 times larger than that measured by the general spectrophotometer In addition the instrument is small outline size 200mmx170mmx110mm and light net weight 1 35kg Principle The calculation of sample concentration is based on the Beer Lambert law A bc Where A is the measured absorbance is the molar absorption coefficient b is the path length and c is the concentration Range of application The YSA 400 micro spectrophotometer can measure Nucleic acid the concentration and purity of DS DNA SS DNA and RNA Protein Othe concentration of the protein 1Abs 1mg mL BSA IgG and lysozyme the concentration of protein with protein assay kit Lowry BCA and Bradford and the software can automatically create the standard curve and report directly the concentration of the sample UV VIS the UV VIS full spectrum 200 850nm absorbance scan Cell Culture the concentration of suspended cell cultures Micro Array the concentration of fluorescent dye labeled nucleic acid micro array for it can measure the concentration of the nucleic acid and the dye at the same time 2 Installation 2 1 Port connection Connect the USB port of the YSA 400 micro spectrophotometer to the USB port of computer with the USB connector Connect the adapter to the power plug with the power adapter connector Plug the plug of the adapter into the jack of the YSA 400 micro spectrophotometer 2 2 System requirement
25. surement type select area Nucleic Acid Protein UV VIS Cell Culture Micro Array and Sample amp Test Type and Sample amp Test Type is the supplement for the former five items The user can select the correct measurement type from here based on the samples Nucleic Acid Protein UV VIS Cell Culture MicroArray Nucleic Acid This type is designed for measuring the concentration and purity of nucleic acid The user can select the DS DNA 50 SS DNA 33 or RNA 44 under the Sample amp Test Type to measure the double stranded DNA the single stranded DNA and RNA respectively 15 Sample amp Test Type Protein This type is designed for measuring the concentration of protein Click the Protein button and enter the protein measurement interface The user can select a specific type from the Sample amp Test Type where are eight types A280 1 BSA 6 7 IgG 13 7 Lysozyme 26 4 Labels BCA Bradford and Lowry Sample amp Test Type A280 1 is designed for measuring the protein solution which produce an absorbance at 280nm of 1 0A path length 10mm at the concentration of 1 mg ml BSA 6 7 IgG 13 7 and Lysozyme 26 4 are designed for measuring the BSA IgG and Lysozyme BCA Bradford and Lowry are designed for measuring the concentration of proteins with protein assay kit A prog
26. tructions of BCA Bradford and Lowry for the experimental principle and preparing for standard proteins and samples These methods need measure the standard proteins and generate a standard curve before measuring the samples Then the concentration will be calculates and reported when a samples is measured In this method the concentration of the sample must be in the concentration range of the standard proteins 5 3 1 Sample size requirements Recommended volume 2 0uL 5 3 2 Measurement range BCA regular 0 2 8 0mg mL mini 0 01 0 2mg mL Bradford regular 0 1 8 0mg mL mini 0 015 0 1mg mL Lowry 0 2 4 0mg mL Typical reproducibility SD mg mL CV BCA Sample range 0 01 0 2mg mL 0 1mg mL Sample range 0 2 8 0mg mL 2mg mL Bradford Sample range 0 015 0 05mg mL 0 004mg mL Sample range 0 05 0 125mg mL 5 Sample range 0 1 0 5mg mL 0 025mg mL Sample range 0 5 8 0mg mL 5 Lowry Sample range 0 2 4 0mg mL 2mg mL 26 5 3 1 Work interface Click the Protein button in the measurement types select area and enter the protein measurement interface Measurement type Click the Sample amp Test Type to pop out a drop down list box where BCA Bradford and Lowry are designed for measuring protein with protein assay kits methods Standard curve These methods need make standard curve before measuring samples The Sample and Standard buttons will appear above Sample amp Test
27. user may also input the max value of the Y axis from the Auto Scale Input dialogue which will appear when click the Auto Scale Max value of Y axis 1 0 0 Set Cancel Multi Draw can let several scan curves resulted from several measurements appear in the graph display area at the same time So the user can compare the reproducibility of the same sample or the changes of different samples Show Grid can let the back ground grid appear or cancel mm Absorbance a bad fod w b a fa fea 5 amp amp F FOE EF 6 EE SB OE E En Eig E piii Worre Lengthinm Click Color Solution a Form Color Solution dialog box will appear From here the user can change the colures of graph display area such as the background the measurement lines and the scan curves And the color of measurement lines and scan curves should be different from the color of background ForaColorSolution Back Ground MicBlck Grid E csive v Axis MicBlack Number MicBlck xb E celow v xc oO cl qua Cancel Tool The Check Calibration and Dye Editor submenus are under the Tool File E View Teel 2 Help H 5 Cheek Calibration Ctrl C Dye Editor Ctrl D Click Check Calibration to pop out a Check Calibration dialog box which enables the user to check whether the light intensity is appropriate The light intensity has been adjusted to the pe
28. wave lengths are the values of Xa Pos Xb Pos and Xc Pos in the data display area respectively and the corresponding absorbances are the values of Xa Value Xb Value and Xc Value respectively Moving the mouse pointer nearby the measurement lines the mouse pointer will change into a double headed arrow When press the left mouse button down and move the mouse pointer the measurement line will move too The measurement lines will move to the corresponding place and the absorbances will also change when the values of Xa Pos Xb Pos and Xc Pos are changed An absorbance curve will appear in the graph display area after measuring a sample Tow curves will appear when both of the wavelengths 1mm and 0 2mm are chosen using UV VIS full spectrum to scan a sample 4 4 Data display area This area displays the wave length absorbance concentration of sample and others as below 17 Xa Pos Xb Pos Xc Pos 260 280 Xa Value Xb Value Xc Value ng ul The absorbances of X Value correspond to the values of X Pos and the vertical measurement lines in the graph respectively The ratio of absorbance at 260nm and 280nm the bar of 260 280 is used to assess the purity of nucleic acid Sample concentration in ng uL is shown in the ng uL bar for nucleic acid and in mg mL for protein when a sample is measured 4 5 Measurement function area There are five buttons in the measurement function area measure Blank Save Graph
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