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Hoefer Vision Life Spectrophotometer User Manual

1. Dilution Factor Yolume Parameters screen or press Back to cancel selections Step 8 Select units of measurement using left and right arrows Options Diluent are ug ml ng ul ug ul Press the down arrow Step 9 Enter the factor using the keypad numbers Range 0 001 to ci Press OK O to enter the results screen or Cancel Q to return to the Applications Folder Version 1 0 Page 25 OOo 000 Absorbance Ratio Concentration 0 26 pal ml Parameters Print Graph Sample Humber Save Method Auto Frint ye e Version 1 0 Results Screen Step 10 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 11 Insert sample and press 0d Repeat step 11 for all samples The absorbance at selected wavelengths is measured and the ratio between wavelengths 1 and 2 is calculated both corrected by the background wavelength value if this was selected Press to return to the Applications Folder Press II to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Graph shows a wavescan plot across the selected wavelengths in place of the individual wavelength 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the
2. Step 2 Enter the first wavelength using either the number keys or the left and right arrows Press the down arrow Enter the second wavelength as above and repeat for the number of wavelengths selected up to 5 Step 3 Press OK Y to enter the results screen OR Press Cancel to return to the Applications Folder Step 4 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 5 Insert sample and press 0d Repeat step 5 for all samples Results A scan plot covering the range of wavelengths selected with cursors at the relevant wavelengths and a table of values is displayed Press to return to the Applications Folder Press II to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above z iS a 2 Print result via selected method es 4 Print graph using selected method Grayed out if no data are available SP Print Graph 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value gt 8 Save method use the left and right arrows to select a folder Ed Sample Number e to store in Favorites Methods 1 9 press the down arrow E Save Method e and enter name E suto Print 9 Auto print toggles auto print on off Exit options by pressing or wait Version 1 0 Page 24 7 Absorbance Ratio This ma
3. Hoefer Where Electrophoresis Gels Vision Spectrophotometer USER MANUAL Hoefer Where Electrophoresis Gels Declaration of Conformity This is to certify that the Vision UV Visible Spectrophotometers part numbers SP 2001 SP 2001PT SP 2001BL and SP 2001SD manufactured by Hoefer Inc conform to the requirements of the following Directives 73 23 EEC amp 89 336 EEC4 IVD Standards to which conformity is declared EN 61010 1 2001 Safety requirements for electrical equipment for measurement control and laboratory use EN 61326 2 3 1998 Electromagnetic compatibility generic emission standard Electrical equipment for measurement control and laboratory use EN 61000 4 6 1992 Electromagnetic compatibility generic immunity standard part 1 Residential commercial and light industry BS EN 591 2001 Instruction for use for in vitro diagnostic instruments for professional use BS EN 13612 2002 Performance evaluation of in vitro diagnostic medical devices 2002 96 EC This appliance is marked according to the European directive 2002 96 EC on Waste Electrical and Electronic Equipment WEEE By ensuring this product is disposed of correctly you will help prevent potential negative consequences for the environment and human health which could otherwise be caused by inappropriate waste handling of this product The symbol on the product or on the documents accompanying the product indicates that this appliance may not
4. Insert sample and press gt Page 12 Concentration Step 6 if using standard mode Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Press 0 to display the Run Standard screen Absorbance Run the standard by pressing 0 OR Press cancel Q to return to the measure screen umol wavelength BO nm Concentration Step 7 Insert the sample and press OF The concentration of the sample is displayed Results shown as indicate the concentration is out of range wavelength BO nm Z 0 042 A Repeat step 7 for all samples Press to return to the Applications Folder Press II to display available Options which are described pmol below Options select using key pad numbers 1 Return to parameters screen step 1 above dd Parameters 2 Print result via selected method O Fin 3 Toggles on off displaying a graph of wavescan 20 nm E Graph from selected wavelength GH Run standard 4 Return to Run Standard screen 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value Ed Sample Number 8 Save method use the left and right arrows to select a folder Save Method to store in Favorites Methods 1 9 press the down arrow E Auto Print wf and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Version 1 0 Page 13 3 Wavescan An absorp
5. Press OK O to accept the calibration and go to the Results a Screen see below OR Press Back Q to return to the Standards screen Results screen Step 13 Insert the reference and press the 0A 100 T key This will be used for all subsequent samples until changed Step 14 Insert the sample and press OF The concentration of the sample is taken and displayed Repeat step 14 for all samples Press to return to the Applications Folder Press II to display available Options which are described below Version 1 0 Page 22 OOo 008 Parameters Print Graph Sample Humber Save Method Auto FPrint cl Version 1 0 Options select using key pad numbers 1 2 3 e 8 9 Return to parameters screen step 1 above Print result via selected method Toggle graph on off Displays calibration graph cursors give values for last measured sample Sample number add a prefix to the sample number and reset the incrementing number to the desired value Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name Auto print toggles auto print on off Exit options by pressing or wait Page 23 6 Multiple Wavelength This makes up to 5 absorbance measurements on the same sample The procedure is as follows Multi wavelength Parameters Step 1 Select the number of wavelengths Press the down arrow
6. Replicates Curve Fit Regression Calibration Replicates Std 4 20 Las mil Std 5 1 00 ua mil Std 6 1 40 pa ml Step 1 Press 3 to select Bradford method Step 2 Wavelength for this stored method is pre set to 595 nm Step 3 Enter the number of standard concentration points 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key BI kdk and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l mol l g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK 0 to store the chosen parameters or Cancel O Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin interpolated or cubic spline Press the down arrow Step 6 Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates usin
7. or wait Page 43 4 Lowry The procedure is as follows Lowry Parameters T50 nm Standards Lowry Parameters Curve Fit Regression Calibration Replicates Standards Calibration ae Units Replicates on Lowry Standards Std 1 Std Std 2 Std 5 0 40 wom 1 00 ua mil Std 3 Std 6 0 60 pa ml 1 40 pa ml Version 1 0 Step 1 Press 4 to select Lowry method Step 2 Wavelength for this stored method is pre set to 750 nm Step 3 Enter the number of standard concentration points 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key Kk and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from O to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK 0 to store the chosen parameters or Cancel Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin interpolated or cu
8. wavelength Curve Fit Step 1 ero Regression Select the wavelength using the keypad numbers or left and right arrows ibid Press the down arrow Step 2 Enter the number of standard concentration points to be used in the curve 1 9 Press the down arrow Replicates Step 3 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key BI kdk and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol l g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters or Cancel O Step 4 Select the type of curve fit using the left and right arrows Standard Curve Parameters Options straight line regression a zero regression this forces the straight line through the origin interpolated or cubic spline wavelength Curve Fit Step 5 KERNA Select the calibration mode either Standards measure prepared standards or Manual keypad data entry Press the down arrow Step 6 if standards has been selected in step 5 Select the number of standards to be measured and averaged at each standa
9. and clear the last digit entered OR Step 3 calculate dilution factor Press Kk to enter the dilution factor screen Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press lt gt to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Enter co efficient 1 280 nm using the keypad numbers Default value is 1 55 range is 1 00 to 9999 Press the down arrow Step 6 Enter co efficient 2 260 nm using the keypad numbers Default value is 0 76 Range is 1 00 to 9999 Press the down arrow Step 7 select the units of measurement using the left and right arrows Options ug ml ng ul and ug pl Step 8 Press OK 0 to enter the Results screen OR Cancel Q to return to the Protein folder Page 36 00 DoS Version 1 0 Protein UW Parameterz Print Graph Sample MHumber Save Method Auto Print Results Screen Step 9 Insert the reference Press 0A 100 T Key This will be used for all subsequent samples until changed Step 10 Insert sample and press 04 This measures at both 260 and 280 nm wavelengths and displays the result Protein conce
10. be treated as household waste Instead it shall be handed over to the applicable collection point for the recycling of electrical and electronic equipment Disposal must be carried out in accordance with local environmental regulations for waste disposal John Attwood Director of Sales amp Marketing Hoefer Inc Hoefer Inc 84 October Hill Road Holliston MA 01746 Toll Free Phone Fax E mail 800 227 4750 1 508 893 8999 1 508 429 5732 support hoeferinc com Website www hoeferinc com Version 1 0 Page 2 TABLE OF CONTENTS ESSENTIAL SAFETY NOTES oa ei 4 Unpacking Positioning and Installation ococcconnconnnocnnocnnocnnecanenarenanonarenarenarennronnrenarennrenarenarenarenanenars 4 INTRODUCTION 0 A A A AAA 5 OU SPEcIroDNOtOMEtO sra A Aa 5 Sample handing ODS A A AA 5 Key pad and CIS Dl Vu A ia 6 SONWare SUIS aa a E E reeneeusueees 8 THE APPLICATIONS FOLDER ci 9 1 Single Wavelength Abs and VT c occcocncccocccccoconoconacoconacononoccnnnnonnnnronnnnrrnnnronnnnrrnnnnrrnnnnrrnnnreranarerannnrrnnans 10 Li CONnC nNtratio Msia EEE EA N aiaa erR N Riia 12 S VVAVES o Y y POCO POE OO Era GEE Nia 14 4 SIMPLE AMOS A in 17 5 SlAMGAlG UI an 20 6 Multiple Wavelength iii ria a a 24 71 ADSODANCO Ral Oia AAA Acida 25 THE LIFE SCIENCE FOLDER cmi Unica 27 O A RR ne Nn 29 A RC a AN 31 O A A A A A ee ee 39 Protem Determination A A ANS 35 A tos eee nee ne 36 y as ean ws ee at eee ene uae
11. been installed it enables the user to print through the PC directly to the printer that is connected to it The data may also be stored as an Excel spreadsheet as an EMF graphics file a comma delimited csv data file a tab delimited txt data file or in native PVC format for later access Alternatively results may be sent to the PC via a Bluetooth accessory this can either be supplied pre installed or is available as an optional accessory if the need for its use arises after installation of the product PVC works in a similar way A printer is available for the instrument this may either be supplied pre installed or is available as an optional accessory if the need for its use arises after installation of the product Sample handling tips e Note that the light beam is directed from RIGHT to LEFT through the cell chamber therefore please ensure the cell is inserted in the correct alignment e The cell holder supplied with the instrument accepts standard 10 mm pathlength quartz glass or plastic cells e The optical beam height is 15 mm and the minimum volume that can be used is approximately 10ul in a Quartz ultra micro cell e 12mm test tubes may be used e g for cell cultures however they are not recommended as higher quality data is produced by using disposable cuvettes for the analysis If used align the indicator line on 12 mm test tubes in the same direction to ensure reproducible positioning of the tube Note that test tub
12. can compensate for dilution and use of cells which do not have 10 mm pathlength by entering dilution factor and cell pathlength Nucleic Acid Purity Checks e Nucleic acids extracted from cells are accompanied by protein and extensive purification is required to separate the protein impurity The 260 280 ratio gives an indication of purity it is only an indication however and not a definitive assessment Pure DNA and RNA preparations have expected ratios of gt 1 8 and 2 0 respectively deviations from this indicate the presence of impurity in the sample but care must be taken in interpretation of results e The 260 nm reading is taken near the top of a broad peak in the Absorbance spectrum for nucleic acids whereas the 280 nm reading is taken on a steep slope i e small changes in wavelength cause large changes in absorbance Consequently small variations in wavelength at 280 nm will have a greater effect on the 260 280 ratio than variations will at 260 nm Thus different instruments may give slightly different ratios due to variations in wavelength accuracy however each instrument will give consistent results within itself e Concentration also affects 260 280 readings If a solution is too dilute the readings will be at the instrument s detection limit and results may vary as there is less distinction of the 260 peak and 280 slope from the background absorbance This is one reason why the Abs260 value should be greater than 0 1 for accu
13. e PVC Print Via Computer is a small application running under Windows 2000 or Windows XP to enable a Vision spectrophotometer to transfer data into a PC environment From there the user has a selection of choices the data can be both printed or saved in a variety of formats PVC is capable of supporting several instruments simultaneously limited only by hardware and the speed of the host system e PVC can operate via USB and Bluetooth simultaneously PVC can store data either to a common directory or be configured to save to independent directories by both file format and connection e PVC can save data in graphics format text format or as an Excel file Installation See the manual included on the PVC CDROM for installation and operating instructions Version 1 0 Page 64 ACCESSORIES USB cable spare source locally Built in printer accessory 80 3003 84 Bluetooth accessory 80 3003 96 SD Card accessory 80 3005 00 MAINTENANCE When using calibration standard filters insert such that the flat surface is facing away from the spring end of the cell holder Observe all necessary precautions if dealing with hazardous samples or solvents Lamp Replacement The Xenon lamp should not need replacement until after several years of use In the unlikely event that it does need replacing this should be undertaken by a service engineer from your supplier Cleaning and general care of the instrument External cleaning Switch off th
14. eaeacies ees oe 38 o A e ate ee eed oe oa 41 A MONI RR 44 9 BUG A eee ncanas aa E setae a 47 Bacterial Cell Culture Measurement OD600 coooooccconncccocccococccoonccononanonnnnnconncnnnnnrnnnnnrrnnnrrnannrrnnnnrrnannrenaness 50 FAVORITES AND METHODS FOLDERS sannansensessensessensensensensunnnnsnnnnnnnnnnnnansansensensensensansensensenssnsnnsnnsnnnnnesansansennee 52 UTILITIES FOLDER ira 53 A A OO 54 DAA eE O Qro AI 54 A A A Bade 54 A A A O A AA 54 da dE y CS OPERA COMPE O A a O mate aed iaaea tals 55 SA A A A A A AA 55 AN A A OA 55 A ER 56 SAME na 56 ACCESSORIES INSTALLATION ici as 58 Printer IMSTAMAUION caca lo aldea 58 Loading changing the printer paper occcocnconccincccconccococonnnnonanonnnrcnnnrennnrennrrnnnrnnrrnnn nena rrnnnrrnnrrnnnrrrnanenannens 60 Bluetooth accessory Installation is 61 PRINTVIA COMPUTER asian 64 ACCESSORIE S naaa 65 Lamp Replacement sarita 65 Cleaning and general care of the insStruMenNt occonncconnnonacincnnoncnnonnnnnnnronnnronaronnnrnnnnrrnnnrnnnrrnnn nena rennrennnnnnns 65 SPECIFICATION AND WCARRAN Ts AA OA aA E EAA ARAN AA ai 66 Version 1 0 Page 3 ESSENTIAL SAFETY NOTES There are a number of warning labels and symbols on your instrument These are there to inform you where potential danger exists or particular caution is required Before commencing installation please take time to familiarise yourself with these symbols and their meaning AN Caution refer to accompanying documen
15. lt gt Peak Detection d E 5 Delete Peak Er E E Graph Scale T E 460 480 500 E Sample Humber E savemenos n EA O O g Auto Print ao 0 020 A A z Pe mum A 496nm Abs 0 020 5 Graph Scale Zoom Mode Hip aHes Version 1 0 Add Peak Shortcut button 5 Adds a used defined peak at the current cursor position The entry is then displayed in inverse coloring to discriminate between user defined peaks and auto detect peaks When the cursor is positioned over the user defined peak a legend User Defined Peak appears at the top of the graph The option then changes to Delete Peak to enable the user to remove the peak Note Storing a method at this stage will save these user defined wavelengths each time method is run Absorbance value at these wavelengths is reported Graph Scale This enables the user to set up a defined graph by defining the limits in either or both of the x and y axes Zoom mode This sets up the operation of the Zoom keys up and down arrows x amp y axes expands the display around the cursor measurement point whilst the other options select the absorbance or wavelength axes respectively With x or y axis limits set to on zooming out will only be permitted to the set limits x y axis limits Setting x or y axis limits to On activates the start and finish points of the desired graph to user defined specific wavelengths and or absorbance values Pressing Canc
16. return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Select the units of measurement using the left and right arrows Options ug ml ng ul ug pl Press the down arrow Step 6 Enter the factor using the keypad numbers Default value is 50 range is 0 01 to 9999 Step 7 Press OK O to enter the Results screen OR Cancel Q to return to the Nucleic Acids folder B ackground Results Screen Step 8 Insert the reference Press 0A 100 T Key This will be used for all subsequent samples until changed Step 9 Insert sample and press This measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 Absorbencies are calculated both corrected by the background wavelength value if selected Gives concentration based on Absorbance at wavelength 1 DNA Repeat step 9 for all samples Press Q to return to the Nucleic acid folder Press II to display available Options which are described below Version 1 0 Page 29 00 000 Version 1 0 Parameterz Print Graph Sample MHumber Save Method Anuto Print Options select using key pad numbers 1 Z 3 Return to parameters screen step 1 above Print result via selected method Toggle graph on off The graph shows a wavescan plot across the range 220 nm to 320 nm with cursors denoting
17. return to the Favorites Methods folder Version 1 0 Page 52 UTILITIES FOLDER of ef ef Oate and Time Regional Printer eo ef Contrast Folder Mames oQ Gamez Summary Function Keypad number Description O 1 Set correct time and date Date and Time e 2 Select preferred language and number format Regional 3 Printer output options Printer 4 Select screen layout themes and history Preferences 5 Adjust screen contrast amp brightness Contrast al 6 Re name Method folders Folder Mames i T Serial number and software version About o ES 8 Spectro Blocks Sudoku Version 1 0 Page 53 Utilities 1 Date and Time The procedure is as follows Enter the day using the keypad numbers or left and right arrows Press the down arrow Enter the month as above Press the down arrow Enter the year Minute Press the down arrow Mach O Enter the hour Press the down arrow Date and Time Year Enter the minute Seconds are zeroed when OK is pressed Press OK 0 to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the time 2 Regional sets Language and Number Format The procedure is as follows Regional Select a language Options are French English or Spanish Language German and Italian will be released in the near future Press the down arrow Set the decimal point style Options are o
18. screen step 1 above Print data on the results screen via selected method Print all the data Set the t position starting point for the slope and dA calculation at the current cursor position Value is retained for subsequent samples Set the t position finishing point for the slope and dA calculation at the current cursor position Value is retained for subsequent samples Toggle the calculated slope line on and off Note if any data points enclosed by t and t are beyond the range of the instrument gt 2 5A or lt 0 3A then this option is grayed out Sample number add a prefix to the sample number and reset the incrementing number to the desired value Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name Auto print toggles auto print on off Exit options by pressing or wait Page 19 5 Standard Curve The construction of a multi point calibration curve from standards of known concentration to quantify unknown samples is a fundamental use of a spectrophotometer this instrument has the advantage of being able to store this curve as a method using up to 9 standards To include a zero concentration standard include this in the number of standards to be entered and enter 0 00 for concentration use a reagent blank when required to enter the zero standard The procedure is as follows Standard Curve Parameters
19. 230 260 280 and if background correction selected 320 nm Sample number add a prefix to the sample number and reset the incrementing number to the desired value Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name Auto print toggles auto print on off Exit options by pressing or wait Page 30 2 RNA The procedure is as follows Step 1 Press 2 to select RNA mode Step 2 Select path length using the left and right arrows Options are 5 or 10 mm Press the down arrow Step 3 dilution factor known RHA Parameters Pathlength Units Enter the dilution factor using the keypad numbers Range 1 00 a to 9999 Use the C button to backspace and clear the last digit Dilution Factor Factor entered ss Step 3 calculate dilution factor nae aaa Press HKM to enter the dilution factor screen Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow aa Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press lt gt to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Select the units of measurement using the left and r
20. 7 A 0 056 A 0 15 0 094 4 0 1324 ter 0 169 A 0 205 A 0 2 04 O lt 6 O B Lo l z 1 4 wavelength T50 nm Absorbance 0 095 4 Curve Fit Interpolation 00 DoS Version 1 0 Parameterz Print Graph Sample MHumber Save Method Auto Print Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Range 0 001 to 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes Press OK Y to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen Results screen Step 14 Insert the reference and press the 0A 100 T key This will be used for all subsequent samples until changed Step 15 Insert the sample and press 0d The concentration of the sample is taken and displayed Repeat step 15 for all samples Press Q to return to the Protein Folder Press Kk to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Displays the calibration graph cursors give values for last measured sample 7 Sample number add a prefix to the sample numbe
21. 99 C button backspaces and clears the last digit entered Step 10 Press Next 0 to enter the Calibration screen If there are duplicate or non monotonic increasing entries the unit will beep and highlight the incorrect entry OR Press Back Q to return to the Parameter screen Page 38 BCA Calibration BCA Calibration Standards 0 057 A 0 177 A 0 238 A 0 404 4 0 516 A 0 627 4 olo s T i h Lo a m 0 3 ml sf a BECA Calibration Replicates 0 057 A 0 057 A Version 1 0 la 0 4 0 6 OB Lo l2 1 4 0 4 a n 04 0 6 08 Lo l2 1 4 na 0 4 0 6 OB Lo l2 1 4 Calibration Screen replicates off This shows the calibration values and allows standards to be measured Step 11 Insert the reference sample Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result Repeat step 12 for all standards A graph will display the results and the fitted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 When all standards are measured the OK box appears Press Y to accept the calibration an go to the Results screen see Press Back Q to cancel selections and return to the Standards screen Calibr
22. Abs 320 Abs ratio Abs 260 Abs 320 Abs 230 Abs 320 e f your laboratory has not used background correction before set this option to NO e The use of background correction can remove variability due to handling effects of low volume disposable cells Spectral scan of nucleic acid Abs PURE NUCLEIC ACID POLY dadT waves 26000 Abal FO a fi ki 0 50 i SS Waves240 0 Alte 343 ho 2000 225 0 250 0 rai 300 0 325 0 350 0 Wenalancih Note e absorbance maximum near 260 nm and absorbance minimum near 230 nm e flat peak near 260 nm and steep slope at 280 nm e very little absorbance at 320 nm Version 1 0 Page 28 1 DNA The procedure is as follows Step 1 Press 1 to select DNA mode Step 2 Select path length using the left and right arrows Options are 5 or 10 mm Press the down arrow Step 3 dilution factor known Pathlength Units Enter the dilution factor using the keypad numbers Range 1 00 sic to 9999 Use the C button to backspace and clear the last digit DAA Parameters Dilution Factor Factor entered ch Step 3 calculate dilution factor Press II to enter the dilution factor screen Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press lt gt to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and
23. Enter the time in minutes over which measurements are taken UUSIA This can be a maximum of 60 minutes Step 4 Interval Enter the interval time in seconds between measurements using the left and right arrows Options are 5 10 20 30 or 60 seconds Step 5 Press Next 0 to go to the next parameters screen OR Press Cancel to return to the Applications Folder Kinetics Parameters 7 Mode Kinetics Parameters 2 Screen Delta A Step 6 select the measurement mode using the left and right arrows Delta A change in absorbance over the measurement duration or selected period Final A absorbance at the end of the measurement duration or selected time Slope rate of change of absorbance over the measurement duration or selected period o o Version 1 0 Page 17 Kinetics Parameters 2 Step 7 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key kM and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol l g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK Y to store the chos
24. K Y to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the settinas 5 Contrast Ambient temperature can affect the display This function can optimize the display for local conditions The procedure is as follows Contrast Contrast Adjust the contrast using the left and right arrows Press the down arrow aa Adjust the brightness using the left and right arrows nanines s Press the down arrow Press OK 0 to store the settings and return to the Utilities folder 6 Folder Names This folder allows you to rename the method or favourite folders Folder Names Select the folder you wish to rename using the left and right arrows Press the down arrow Input the new name for the folder Press OK Y to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the settings Folder Hew Hame Version 1 0 Page 55 7 About o Serial Number a E Version 411041 1 Displays the instrument serial number and software version Press OK 07 to close the window and return to the Utilities folder 8 Games Spectro Blacks Sudoku 1 Spectroblocks 1 Hew Game 2 Options 3 Exit Classic block dropping game Follow the instructions Press Cancel to return to the Utilities folder without storing the settings Version 1 0 Page 56 2 Sudoku Can be
25. aneous equations solving these provides the two coefficients In cases where Factor 2 is found to be negative it should be set to zero since it means there is no contribution to the protein concentration due to absorbance at 260 nm Set Factor 2 0 00 for direct A280 UV protein measurement Factor 1 is based on the extinction coefficient of the protein If BSA bovine serum albumin is an acceptable standard setting Factor 1 1 115 will give linear results from 0 to 0 8 mg ml protein Protein mg ml 1 115 Abs 280 Rapid measurements such as this at Abs 280 are particularly useful after isolation of proteins and peptides from mixtures using spin and HiTrap columns by centrifuge and gravity respectively Protein Determination at 595 546 562 and 750 nm The Bradford method depends on quantitating the binding of a dye Coomassie Brilliant Blue to an unknown protein and comparing this binding to that of different known concentrations of a standard protein at 595 nm this is usually BSA bovine serum albumin The Biuret method depends on reaction between Cupric ions and peptide bonds in an alkali solution resulting in the formation of a complex absorbing at 546 nm The BCA method also depends on reaction between cupric ions and peptide bonds but in addition combines this reaction with the detection of cuprous ions using bicinchoninic acid BCA giving an absorbance maximum at 562 nm The BCA process is less sensitive to the presence of deterg
26. ards are measured the OK box appears Press eta even 0 to accept the calibration an go to the Results screen see oe oa e Press Back to cancel selections and return to the Standards 0 60 screen Calibration Screen replicates on 1 40 E This shows the calibration values and allows standards to be measured 1E 04 06 08 1410 Le 1 4 Step 11 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Press Y to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press 0 to measure the standard and store the result Bradford Calibration Repeat for all replicates and standards Replicates 1 0 058 A 2 0 058 A 3 0 058 A A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous 3 reading Step 13 de ou 05 of uo La s Press Y to accept the calibration and go to the Results screen een Press Back Q to return to the Standards screen Version 1 0 Page 42 Bradford Calibration Standards 0 20 0 40 0 60 0 80 0 055 4 0 1964 0 3304 0 4634 D L n 0 4 D6 O B Lo lz 1 4 Bradford wavelength 535 nm 0 331 A Curve Fit Regression 00 000 Version 1 0 Parameterz Print Graph Sample Humber Save Method Auto Pr
27. as been thoroughly tested to ensure that it meets its published specification The warranty included in the conditions of supply is valid for 12 months only if the product has been used according to the instructions supplied The supplier can accept no liability for loss or damage however caused arising from the faulty or incorrect use of this product e This product has been designed and manufactured by Hoefer Inc 84 October Hill Road Holliston MA 01746 However please contact your dealer if you experience technical or sample handling difficulties Version 1 0 Page 66
28. ation Screen replicates on This shows the calibration values and allows standards to be measured Step 11 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Press Y to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press to measure the standard and store the result Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 Press Y to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen Page 39 BCA Calibration Standards O m a hte ee polo mia eleisia eia 0 087 A 0 1714 0 5 0 238 A 0 404 A 0 3 0516 A 06274 E pE l n 0 4 D E DE 140 142z 14 O E BCA wavelength 562 nm 0 259 4 Curve Fit Regression 00 000 Version 1 0 Parameterz Print Graph Sample MHumber Save Method Anuto Print Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Ran
29. bic spline Press the down arrow Step 6 Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates using the left and right arrows This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2or3 Step 8 Press Next O to enter the Standards screen OR Press Cancel Q to cancel selections and return to the Protein folder Standards Screen Step 9 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears the last digit entered Step 10 Press Next 0 to enter the Calibration screen If there are duplicate or non monotonic increasing entries the unit will beep and highlight the incorrect entry OR Press Back to return to the Parameter screen Page 44 Calibration Screen replicates off This shows the calibration values and allows standards to be Lowry Calibration measured ss aaa ozo Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result T aian ea aai si Repeat step 12 for all standards A graph will display the results and the fi
30. current Enter O Version 1 0 wavelength in the mode selected When in scan mode do a reference scan Enter or confirm a selection Take a measurement Page 6 toleo luoteta ir Parameters Print Print Data Set tO At Cursor Set tn At Cursor Slope Sample Humber Save Method Auto Print wv bi Version 1 0 Options select using key pad numbers 1 View parameters for the experiments 2 Print the results 3 4 5 6 Described in the application T Define the sample number you wish to start from 8 Save the parameters as a method to a defined folder name with a defined method name 9 Toggle auto print on off Default is off Exit options by pressing or wait Experienced operators can use the numeric keys as a shortcut to the option required without needing to enter the Options menu Page 7 Software style The user interface is built around having folders of files which are displayed on the home page when the instrument is switched on Different folders are numbered and opened by using the associated number key on the keypad Hoefer Vision Spectrophotometer el Applications Favourites Methods oQ eQ Utilities Life Science Summary Function Keypad number Description o ma 1 Single wavelength Concentration Wavelength scan Kinetics Standard Curve Multiple wavelengths and Ratio Application a 2 saved User selected and configured methods e ah Favourites gt 3 Sub folde
31. e instrument Note that setting the History parameter to on see Preferences later will cause the instrument to store it s last settings If the History parameter is turned off all parameters and options will return to their default settings when you leave that application Unless it has been saved as a method Version 1 0 Page 9 1 Single Wavelength Abs and T This makes simple Absorbance A and Transmission T measurements on samples measuring the amount of light that has passed through a sample relative to a reference this can be air The procedure is as follows Single Wavelength Parameters Step 1 Set wavelength by using keypad numbers or left and right wavelength arrows Press the down arrow key Step 2 wlll Select the mode Absorbance or T using the left and right arrows Step 3 To enter the results screen with the selected parameters press ce OR Cancel the selections and return to the Applications Folder by pressing Cancel O Single wavelength Step 4 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 5 Insert sample and press 0d Repeat step 5 for all samples Results The result at the selected wavelength is displayed on screen Use the left and right arrows to move the cursor and display the value at the cursor position 15nm from set wavelength Press Cancel to return to the Applications Folder Press K
32. e instrument and disconnect the power cord Use a soft damp cloth Clean all external surfaces A mild liquid detergent may be used to remove stubborn marks Changing cell holder or removal for cleaning This can be removed by undoing the appropriate screws on the bottom of the instrument Version 1 0 Page 65 SPECIFICATION AND WARRANTY Wavelength range Monochromator Wavelength calibration Spectral bandwidth Wavelength accuracy Wavelength reproducibility Light sources Detector Photometric range Photometric linearity Photometric reproducibility Stray light Zero Stability Noise Digital output Dimensions Weight Power input 190 1100 nm Flat grating Automatic upon switch on 5 nm 2 nm 1 nm Pulsed xenon lamp 1024 element CCD array 0 300 to 2 500A O to 199 T 0 005 Abs or 1 of the reading whichever is the greater 546 nm 0 003 Abs 0 to 0 5 Abs 0 007 Abs 0 5 1 0 Abs lt 0 5 at 220 nm and 340 nm using NaNO 0 01 Abs hour after 20 min warm up 340 nm 0 005 pk to pk 0 002 pms USB port standard Bluetooth option 260 x 390 x 100 mm lt 4 5 kg 90 250 V 50 60 Hz Max 30 VA Specifications are measured after the instrument has warmed up at a constant ambient temperature and are typical of a production unit As part of our policy of continuous development we reserve the right to alter specifications without notice Warranty e Your supplier guarantees that the product supplied h
33. ect the mode Factor user entered or Standard factor is calculated from a calibration sample using the left and right arrows Press the down arrow key Step 3 if Factor is selected Enter the Factor using the keypad numbers Range 0 001 to 9999 Use the C button to delete the last digit entered Press the down arrow key Step 3 if Standard is selected Enter the concentration using keypad numbers Range 0 01 9999 Use the C button to delete the last digit entered Press the down arrow key Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key Kk and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from O to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK W to store the chosen parameters or Cancel O Step 5 To enter the results screen with the selected parameters press Cancel the selections and return to the Applications Folder by pressing Cancel Step 6 if using a Factor Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 7
34. el O ignores the selection pressing 0 accepts them and displays the required graph Page 16 4 Simple Kinetics Kinetics studies where the change in absorbance needs to be followed as a function of time at a fixed wavelength can be readily performed Reagent test kits are routinely used for the enzymatic determination of compounds in food beverage and clinical laboratories by measuring NAD NADH conversion at 340 nm The change in absorbance over a specified time period can be used to provide useful information when an appropriate factor defined in the reagent kit protocol is applied Reaction rate and enzyme activity can be calculated if the factor used takes account of the absorbance difference per unit time as opposed to the absorbance difference per se For this reason the change in absorbance per minute AA min concentration AA min x factor and correlation coefficient calculated from a best fit of the data points are displayed They may not be relevant for simple kinetics experiments The procedure to define a new method is as follows Kinetics Parameters 1 Screen Step 1 Wavelength Kinetics Parameters 1 Enter all numerical values using the keypad numbers or the left and right arrows Use the up and down arrow keys to move Wavelength between boxes 0 Seconds Step 2 Delay time Enter the delay time in seconds before measurements are taken Burson This can be a maximum of 600 seconds 10 minutes Step 3 Duration
35. en parameters or Cancel O Step 8 Set the Factor by which the result is multiplied to give the amount in the chosen range using the left and right arrows Range of 0 01 to 9999 Step 9 Press Next O to enter the Results screen OR Press Cancel Q to return to the Parameters 1 screen Factor Results Insert the reference and press the 0A 100 T key Kinetics Insert the sample and press 0 to start the run Time min is displayed at the bottom of the screen and absorbance data are plotted on the graph as testing proceeds The table below the graph gives Absorbance values at T start of calculation T finish of calculation change in absorbance slope regression parameter RS of the calculated slope and the result calculated from the selected parameter dA final A or 2 0 0 0 4 0 6 0 8 1 0 o an an stove wa A one Sample 1 Time 00 00 10 Abs 0 231 Use the left and right arrows to move the cursor and display the time and absorbance value at measured data points Use the up and down arrows to zoom in or out Press Cancel to return to the Applications Folder Press kk to display available Options which are described below Version 1 0 Page 18 efollofo le lolrio Parameters Print Print Data Set tO At Cursor Set tn At Cursor Slope Sample Humber Save Method uto Print e Version 1 0 Options select using key pad numbers 1 2 3 4 9 Return to parameter 1
36. ents used to break down cell walls The Lowry method depends on quantifying the color obtained from the reaction of Folin Ciocalteu phenol reagent with the tyrosyl residues of an unknown protein and comparing with those derived from a standard curve of a standard protein at 750 nm this is usually BSA bovine serum albumin Detailed protocols are supplied with these assay kits and must be closely followed to ensure accurate results are obtained The use of plastic disposable cells is recommended To use a zero concentration standard include it in the number of standards to be entered and enter 0 00 for concentration use this when required to enter standard 1 A linear regression analysis of the calibration standard data points is calculated the result together with the correlation coefficient can be printed out A correlation coefficient of between 0 95 and 1 00 indicates a good straight line Version 1 0 Page 35 1 Protein UV This is the Christian and Warburg assay discussed previously The procedure is as follows Protein UV Parameters Pathlength Coeff 1 1 55 Dilution Factor Coeff 2 1 00 0 re Background Units was ml Version 1 0 Step 1 Press 1 to select Protein UV mode Step 2 Select path length using the left and right arrows Options are 5 or 10 mm Press the down arrow Step 3 dilution factor known Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace
37. es do not last forever and that the surface becomes scratched and blemished through repetitive use if this is the case they should be replaced Version 1 0 Page 5 Keypad and display The back lit liquid crystal display is very easy to navigate around using the alphanumeric entry and navigation arrow keys on the hard wearing spill proof membrane keypad 45 5 EN ghi jh kma as Alphanumeric Keys i CTN G 3 vq ES e NES Dis H YC A om Escape Cancel eee Set Reference Arrow Keys Options Key a A Enter selection Run measurement Key Action On off key Turns the instrument on off Arrow keys Use the four arrow keys to navigate around the display and select the required setting from the active highlighted option View Options Bd B k View options for that application mode Some of these are common to all applications and described below Options unique to an application are described in the relevant section Alphanumeric keys Use these to enter parameters and to write text descriptions where appropriate or required Use repeated key presses to cycle through lower case number and upper case Leave for 1 second before entering next character Use C button to backspace and 1 to enter a space Escape Cancel Escape from a selection and return to the previous folder Stop making measurements Set Reference 0A 100 T Set reference to 0 000 A or 100 T on a reference solution at the
38. ess OK Y to accept the calibration and go to the Results mo m mo 40 so n screen see below OR Press Back Q to return to the Standards screen Version 1 0 Page 21 Calibration Screen replicates on Standard Curve Calibration This shows the calibration values and allows standards to be measured Step 10 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 11 Press 0 to display the replicate entry boxes Use C to clear previously stored results before measuring 0 Insert the standard and press Enter to measure the standard and jogo store the result Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 12 Press 0 to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen Standard Curve Calibration Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad E The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Range 0 001 to 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes
39. ethods 1 9 press the down arrow and enter name Auto print toggles auto print on off Exit options by pressing or wait Page 32 3 Oligo The procedure is as follows Step 1 Press 3 to select Oligo mode Oligo Parameters Step 2 Select path length using the left and right arrows Options are 5 Pathlength Units or 10 mm Press the down arrow Step 3 dilution factor known ae Factor Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace and clear the last digit entered Background OR Step 3 calculate dilution factor Press Kk to enter the dilution factor screen D OK ena Enter the volume of the sample using the keypad numbers Range 0 01 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers Range 0 01 to 9999 Press to calculate the dilution factor and return to the Parameters screen OR Press to cancel the selections and return to the Parameters screen Step 4 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow Step 5 Select the units of measurement using the left and right arrows Options ug ml ng ul ug ul and pmol ul If pmol ul is selected the factor changes to a selection table denoting the ratios of the 4 bases in the structure Press the down arrow Step 6 units not pmol ul Enter the factor using the keypad numbers Default
40. ext string up to 8 characters long To access a list of pre defined units press the Options key III and then use the left right arrows ug ml ug pl pmol ul mg dl mmol l umol l g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK IS pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK 0 to store the chosen parameters or Cancel Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin interpolated or cubic spline Press the down arrow Step 6 Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates using the left and right arrows This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2 or 3 Step 8 Press Next O to enter the Standards screen OR Press Cancel Q to cancel selections and return to the Protein folder Standards Screen Step 9 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 99
41. f Exit options by pressing or wait Peak Detection Shortcut button 4 AutoDetect Peaks Turns on and off the automatic peak detection The following options determine how peaks are detected Minimum peak height Minimum height the peak has to be above the higher of the two adjacent minima for the peak to be detected Minimum peak width Minimum width of the peak as determined by the difference in wavelength between the higher of the two adjacent minima and the opposing intersection of that higher minimum level and the peak profile See the screen displayed below Peak Detect on Zoom Determines whether peaks are re assessed and tabulated when the user zooms into a region of the wavescan If off leaves the peak detection as determined on the un zoomed display Sort peaks by Determines the sequence that peaks are reported by Can be wavelength peak height or peak width Draw Peaks Switches display of peak cursors on and off These show vertical dashed lines displaying the measured peak height and horizontal dashed lines showing the peak width Pressing Cancel ignores the selection pressing Y accepts them Page 15 wW avescan User Defined Peak 0 4 E 400 420 440 460 450 500 a 412 as 468 sor Abs 0 2821 0 517 0 121 0 210 M0 ae eel a Sample 2 A 436nm Abs 0 070 4 wW avescan User Defined Peak 0 5 l 0 4 i l 1 Parameters 0 Pririt i E Abs T po i
42. g the left and right arrows This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2or3 Step 8 Press Next O to enter the Standards screen OR Press Cancel Q to cancel selections and return to the Protein folder Standards Screen Step 9 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears the last digit entered Step 10 Press Next 0 to enter the Calibration screen If there are duplicate or non monotonic increasing entries the unit will beep and highlight the incorrect entry OR Press Back to return to the Parameter screen Page 41 Calibration Screen replicates off This shows the calibration values and allows standards to be Bradford Calibration measu red Si Step 11 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear previously stored results before measuring Press to measure the standard and store the result Repeat step 12 for all standards A graph will display the results and the fitted curve as the measurements are made n fia 0 4 0 6 08 Lo ld 1 4 Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 When all stand
43. ge 0 001 to 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes Press OK 0 to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen Results screen Step 14 Insert the reference and press the 0A 100 T key This will be used for all subsequent samples until changed Step 15 Insert the sample and press The concentration of the sample is taken and displayed Repeat step 15 for all samples Press Q to return to the Protein Folder Press Kk to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Displays the calibration graph cursors give values for last measured sample 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 40 3 Bradford The procedure is as follows Bradford Parameters Bradford Parameters 555 nm Standards Bradford Standards Std 2 0 40 wom Std 3 0 60 pa ml Version 1 0 Curve Fit Regression Calibration Standards
44. graph on off Displays the calibration graph cursors E Graph give values for last measured sample 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder EY Sample Number to store in Favorites Methods 1 9 press the down arrow E Save Method and enter name E Auto Print 9 Auto print toggles auto print on off Exit options by pressing or wait Version 1 0 Page 49 Bacterial Cell Culture Measurement OD600 e Bacterial cell cultures are routinely grown until the absorbance at 600 nm known as OD600 reaches approximately 0 4 prior to induction or harvesting A linear relationship exists between cell number density and OD 600 up to approximately 0 6 e tis important to note that for turbid samples such as cell cultures the absorbance measured is due to light scattering and not the result of molecular absorption The amount of scatter is affected by the optics of the system distance between the cell holder and instrument exit slit geometry of this slit and the monochromator optics Different spectrophotometer types therefore give different responses for the same turbid sample to compare results they must be normalised using calibration curves e A calibration curve can be determined by comparing measured OD 600 to expected OD 600 Expected OD 600 is determined by counting cell number using an a
45. he Graph Scale option and this is retained for subsequent measurements To zoom out again use the down arrow Press to return to the Applications Folder Press XII to display available Options which are described next Page 14 Parameters Print Abs 387 Peak Detection Add Peak Graph Scale Sample Mumber Save Method Auto Print 6068606006 Peak Detection AutoDetect Peaks Peak Detect on Zoom Minimum Peak Height Sort Peaks By ICE OO wa Minimum Peak width Draw Peaks pcm o x cme MW avescan 0 5 0 4 Version 1 0 Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle between Absorbance and T mode 4 Displays Peak Detection Parameter Screen See description below 5 Manually adds a peak position to the peak table in the results screen at the position set by the cursor If the cursor is returned to this position the legend User Defined Peak is displayed at the top of the scan and this option changes to Delete Peak 6 Displays Graph Scale Parameter Screen See description below 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on of
46. he down arrow Step 5 if cells ml selected se edi sie Select the multiplier using the left and right arrows Options are 1000 or 1 000 000 Step 6 Units Press OK lt gt to enter the Results screen OR Press Cancel Q to cancel selections and return to the Life Science folder Version 1 0 Page 50 wavelength B00 nm Absorbance 0 0025 Go 008 Version 1 0 Parameters Print Sample Humber Save Method Auto Print Results Screen Step 8 Insert the reference and press the 0A 100 T key This will be used for all subsequent samples until changed Step 9 Insert the sample and press 0d The wavelength absorbance and OD600 value is displayed Repeat step 9 for all samples Press Q to return to the Life Science Folder Press Kk to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 51 FAVORITES AND METHODS FOLDERS These folders are the storage locations for any user modified Applications Methods that are saved in the Op
47. ight arrows Options ug ml ng ul ug pl Press the down arrow Step 6 Enter the factor using the keypad numbers Default value is 40 range is 0 01 to 9999 Step 7 Press OK O to enter the Results screen OR Cancel Q to return to the Nucleic Acids folder PRG Results Screen Step 8 Insert the reference Press 0A 100 T Key This will be used for all subsequent samples until changed Step 9 Insert sample and press This measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 absorbencies are calculated both corrected by the background wavelength value if selected Gives concentration based on absorbance at wavelength 1 Repeat step 9 for all samples Press Q to return to the Nucleic acid folder Press II to display available Options which are described below Version 1 0 Page 31 00 008 Version 1 0 Parameterz Print Graph Sample Humber Save Method Anuto Print Options select using key pad numbers Return to parameters screen step 1 above Print result via selected method Toggle graph on off The graph shows a wavescan plot across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if background correction selected 320 nm Sample number add a prefix to the sample number and reset the incrementing number to the desired value Save method use the left and right arrows to select a folder to store in Favorites M
48. int Calibration Manual entry Shows previously entered calibration values and allows values to be entered via the keypad The highlighted box can be edited in order to enter an absorbance value corresponding to a given concentration value using the keypad numbers Range 0 001 to 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes Press OK 0 to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen Results screen Step 14 Insert the reference and press the 0A 100 T key This will be used for all subsequent samples until changed Step 15 Insert the sample and press 0d The concentration of the sample is taken and displayed Repeat step 15 for all samples Press Q to return to the Protein Folder Press Kk to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off Displays the calibration graph cursors give values for last measured sample 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing
49. k to display available Options which are described below Version 1 0 Page 10 DOS 5000 Parameters Print AbsT Print Graph Sample Humber Save Method E uto Print yl Version 1 0 Options select using key pad numbers Return to parameters screen step 1 above Print result via selected method Toggle between Absorbance and T mode Print graph grayed out if no data are available Sample number add a prefix to the sample number and reset the incrementing number to the desired value Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off eS I Sa Exit options by pressing or wait Page 11 2 Concentration This makes simple concentration measurements on samples by measuring the amount of light that has passed through a sample relative to a reference this can be air Concentration is obtained by multiplying the measured Absorbance at a specific wavelength by a factor The factor may be known in advance or may be calculated by the instrument by measuring a standard of known concentration The procedure is as follows Concentration Parameters wavelength 6 nm Concentration Parameters Concentration 1 40 Version 1 0 Step 1 set wavelength by using keypad numbers or left and right arrows Press the down arrow key Step 2 Sel
50. kes simple Absorbance ratio measurements on samples measuring the amount of light that has passed through a sample relative to a blank this can be air at two wavelengths The procedure is as follows Step 1 SE O O O Enter the first wavelength by using the keypad numbers or the left and right arrows Press the down arrow Step 2 Enter the second wavelength as above Press the down arrow Step 3 Select whether a background correction is applied to both wavelengths 1 and 2 using the left and right arrows Step 4 If background correction is On Enter the third wavelength from which the background correction will be obtained Wavelength 1 Wavelength 2 250 nm Background Step 5 Press Next lt gt to enter the Parameters screen OR Absorbance Ratio Parameters Press Cancel to return to the Applications Folder Pathlength PE Absorbance Ratio Parameters Screen Step 6 Select the pathlength 5 or 10 mm using the left and right Dilution Factor arrows 1 00 Press the down arrow Step 7 Dilution Factor known Units Enter a dilution factor by using the keypad numbers within the wa ral range 1 00 9999 OR Step 7 Calculate Dilution Factor Press the options key BIB M Enter the volume of the sample range 0 01 9999 using the keypad numbers Press the down arrow Enter the volume of diluent range 0 01 9999 by using the keypad numbers Press OK O to calculate the dilution factor and return to the
51. left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 26 THE LIFE SCIENCE FOLDER Life Science Folder This contains three sub folders Nucleic Acids Protein and Cell Count Contents of these sub folders are detailed below Acids Po J2 RNA Concentration and purity check for RNA samples o 3 Olg_____________ Concentration and purity check for oligo samples A eee INIA ea UV protein Christian Warburg Protein determination at 280nm 2 BCA Protein determination at_562nm PF 3 Bradford Protein determination at 595nm Pp Alar Protein determination at_750nm O Biuret Protein determination at_546nm da A AAN IIA ee 3 Cell Count OD600 Cell culture OD600 with correction factor E A DNA RNA and oligonucleotide characterization Nucleic Acid Quantification NAQ e Nucleic acids can be quantified at 260 nm because it is well established that a solution of DNA in a 10 mm pathlength cell with an optical density of 1 0 has a concentration of 50 or 40 ug ml in the case of RNA Oligonucleotides have a corresponding factor of 33 ug ml although this does vary with base composition this can be calculated if the base sequence is known Concentration Abs260 Factor e The instrument uses factors 50 40 and 33 as defaults for DNA RNA and oligonucleotides respectively and
52. lternative technique for example microscope slide method and converting to OD 600 using the rule of thumb that 1 OD 600 8 x 10 cells ml for E Coli e Your Vision spectrophotometer has much smaller optics than most conventional spectrophotometers and more light is transmitted through to the detector resulting in lower than expected OD 600 values Results obtained by comparing measured OD 600 with expected OD 600 see above indicate that a correction factor of 2 0 is required to make the data comparable to larger instruments this factor is included as a default value in set up e The use of 10 mm pathlength disposable cells is recommended for optical density measurements of cell culture solutions to prevent the suspension settling too quickly and giving an OD that changes with time glycerol should be added to the sample The procedure is as follows OD 600 Parameters Step 1 EE Select the wavelength Default value is 600 nm Press the down arrow Step 2 Correction Enter the factor to compensate for different optical configurations 2 00 between this and other instruments Default value is 2 Press the down arrow Step 3 Select the units Options are OD or cells ml If cells ml is selected two further parameters are displayed OD 600 Parameters Step 4 if cells ml selected Enter the factor using the keypad numbers Range 0 00 to 9999 Wavelength Factor C button backspaces and clears the last digit entered 600 nm Press t
53. nt will perform a series of self diagnostic checks e Please read through this user manual prior to use e Please contact your original supplier in the first instance if you experience technical or sample handling difficulties pT If this equipment is used in a manner not specified or in environmental conditions not appropriate for safe operation the protection provided by the equipment may be impaired and instrument warranty withdrawn Version 1 0 Page 4 INTRODUCTION Your spectrophotometer Your spectrophotometer is a simple to use UV Visible instrument with a CCD array detector 1024 pixels It has no moving parts which is the basis of the rapid scanning operating system The user interface is built around folders which are displayed on the home page when the instrument is switched on After switch on and calibration the default home page is Vision offering the choice of Applications General spectroscopic methods Favorites A folder to store your more frequently used configured methods Methods Contains nine folders that can store less frequently used configured methods nine methods per folder Utilities Instrument set up date time language etc and games Life Science Standard Life Science methods such as nucleic acid assays protein assays and cell counting The instrument is supplied with a program PVC Print via Computer on the accompanying CD When used with a USB cable to connect to a PC onto which the software has
54. ntration is calculated corrected by background wavelength value if selected Repeat step 10 for all samples Press Q to return to the Protein folder Press Kk to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off The graph shows a wavescan plot across the range 250 nm to 330 nm with cursors denoting 230 260 280 and if background correction selected 320 nm 7 Sample number add a prefix to the sample number and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 37 2 BCA The procedure is as follows BCA Parameters 562 nm Regression Curve Fit Calibration Standards Replicates BCA Parameters 562 nm Regression Curve Fit Standards Calibration Standards Replicates BECA Standards Version 1 0 Step 1 Press 2 to select BCA mode Step 2 Wavelength for this stored method is pre set to 562nm Step 3 Enter the number of standard concentration points 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 Units The user can enter a t
55. number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 34 Protein Determination Protein Determination at 280 nm Protein can be determined at 280 nm due to absorption by tyrosine tryptophan and phenylalanine amino acids Abs 280 varies greatly for different proteins due to their amino acid content and consequently the specific absorption value for a particular protein must be determined The presence of nucleic acid in the protein solution can have a significant effect due to strong nucleotide absorbance at 280 nm This can be compensated by measuring Abs 260 and applying the equation of Christian and Warburg for the protein crystalline yeast enolase Biochemische Zeitung 310 384 1941 Protein mg ml 1 55 Abs 280 0 76 Abs 260 or Protein conc Factor 1 Abs 280 Factor 2 Abs 260 This equation can be applied to other proteins if the corresponding factors are known The instrument can determine protein concentration at 280 nm and uses the above equation as default the factors can be changed and the use of background correction at 320 nm is optional To customize the equation for a particular protein the absorbance values at 260 and 280 nm should be determined at known protein concentrations to generate simple simult
56. r Humber Format Press OK O to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing the settings 3 Printer Sets up printing options The procedure is as follows Select whether auto print is on or off using the left and right arrows When auto print is on the results are automatically Auto Print printed after a measurement is taken When it is off printing has to be initiated manually This can also be set using the Options key KHK in each application or method The default is OFF Printer Press the down arrow Select how the data are sent Options are Built in internal printer SD Card accessory or to a computer via USB port or Bluetooth Press OK 0 to store the settings and return to the Utilities folder OR Press Cancel to return to the Utilities folder without storing Printer Version 1 0 Page 54 4 Preferences Sets user preferences The procedure is as follows AA Select games function This determines whether the games folder is displayed or not Options are yes or no Press the down arrow Define the screen layout of folders Options are either a grid format default or a list Press the down arrow Select whether to use previously entered parameters on switch on or use defaults Press the down arrow Select whether to use a standby mode after defined periods Options are 1 hour 2 hours at night or off Press O
57. r and reset the incrementing number to the desired value 8 Save method use the left and right arrows to select a folder to store in Favorites Methods 1 9 press the down arrow and enter name 9 Auto print toggles auto print on off Exit options by pressing or wait Page 46 5 Biuret The procedure is as follows Biuret Parameters Biuret Parameters Curve Fit Regression Calibration Standards Replicates Standards Calibration Units Replicates a Biuret Standards Std 1 Std 0 20 mg di 0 80 mgr dl Std 2 Std 5 0 40 mar dl 1 00 mgr dl Std 3 Std 6 0 60 mar dl 1 40 mas dl Version 1 0 Step 1 Press 5 to select Biuret method Step 2 Wavelength for this stored method is pre set to 546 nm Step 3 Enter the number of standard concentration points 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 Units The user can enter a text string up to 8 characters long To access a list of pre defined units press the Options key BI kdk and then use the left right arrows ug ml ug ul pmol ul mg dl mmol l umol I g l mg l ug l U I ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from O to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal point
58. r selection for user selected and configured methods eL Methods 4 Instrument set up date time language etc and Games o Utilities 5 Nucleic acids Proteins and Cell counting oQ Life Science Version 1 0 Page 8 THE APPLICATIONS FOLDER Applications o e Single wavelength Concentration Mlallezcan E Z o O Opt Kinetics Standard Curse Multi w auelength Ain o Absorbance Ratio SUMMARY Function Key pad number Description o I os 1 Absorbance or T Transmission at a single user defined Single wavelength wavelength e wi Concentration 2 Concentration measurement at a single wavelength based on a simple Factor entered or calculated from a single standard o WaAVeSscan 3 Wavelength scan between two user defined wavelengths Range 200 950 nm with user configurable peak finding function o Ke ees 4 Absorbance versus time measurements either rate or end value based o er 5 Generation of calibration curve by measuring standards at a single wavelength ib 6 Absorbance or T Transmission at up to 5 user defined wavelengths Fiaa o Absorbance Ratio T Ratio of absorbance values at two user specified wavelengths OPTIONS Within each application the user has the possibility to select various options that define the way results are treated If not using a stored method it is advisable to check that these Options have been appropriately set for your experiment when coming to th
59. rate measurements e An elevated absorbance at 230 nm can indicate the presence of impurities as well 230 nm is near the absorbance maximum of peptide bonds and also indicates buffer contamination since Tris EDTA and other buffer salts absorb at this wavelength When measuring RNA samples the 260 230 ratio should be gt 2 0 a ratio lower than this is generally indicative of contamination with guanidinium thiocyanate a reagent commonly used in RNA purification and which absorbs over the 230 260 nm range A wavelength scan of the nucleic acid is particularly useful for RNA samples e The instrument can display 260 280 and 260 230 ratios and compensates for dilution and use of cells that do not have 10 mm pathlength dilution factor and cell pathlength can be entered Version 1 0 Page 27 Use of Background Correction e Background correction at a wavelength totally separate from the nucleic acid and protein peaks at 260 and 280 nm respectively is sometimes used to compensate for the effects of background absorbance The wavelength used is 320 nm and it can allow for the effects of turbidity high absorbance buffer solution and the use of reduced aperture cells The instrument can use background correction e If itis used there will be different results from those when unused because Abs320 is subtracted from Abs260 and Abs280 prior to use in equations Concentration Abs 260 Abs 320 Factor Abs ratio Abs 260 Abs 320 Abs 280
60. rd concentration point Can be OFF 1 2 or 3 cid Step 7 ano Next O to enter the Standards screen Press Cancel Q to cancel selections and return to the Applications Folder Standards Calibration Units Replicates Version 1 0 Page 20 Standard Curve Standards Standards screen Step 8 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 Step 9 Press Next Y to enter the Calibration screen If any duplicate or non monotonic increasing entries are present the unit will beep and highlight the incorrect entry OR Press Back Q to return to the Parameter screen Standard Curve Calibration Calibration Screen replicates off N This shows the calibration values and allows standards to be measured DECINE as Step 10 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 11 Insert the standard use C to clear previously stored results before measuring a do Press to measure the standard and store the result Repeat for all standards Sandara IES aren A graph will display the results and the fitted curve as the measurements are input Standards 0 12 10 0 0 015 A E D 10 00 TEA Use the up and down arrows to select a standard to be repeated 31 0 coco MN if a poor reading has been obtained Use C to clear the previous 62 0 0 06 reading Step 12 Pr
61. s are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK 0 to store the chosen parameters or Cancel O Step 5 Enter the type of curve fit Options are straight line regression zero regression forces the straight line through the origin interpolated or cubic spline Press the down arrow Step 6 Select the calibration mode either standards measure prepared standards or manual keypad data entry Step 7 if standards selected Select the number of replicates using the left and right arrows This determines the number of standards to be measured and averaged at each standard concentration point Can be OFF 1 2 0r3 Step 8 Press Next O to enter the Standards screen OR Press Cancel Q to cancel selections and return to the Protein folder Standards Screen Step 9 Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes Range 0 001 to 9999 C button backspaces and clears the last digit entered Step 10 Press Next O to enter the Calibration screen OR Press Back Q to return to the Parameter screen Page 47 Calibration Screen replicates off This shows the calibration values and allows standards to be measured Biuret Calibration Step 11 2 5 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Insert the standard use C to clear pre
62. set up as Computer mode 50 preset games or User mode enter your own pattern Use the cursors to select the square and the key pad to enter a number Invalid numbers cannot be entered Cells can be locked or unlocked by using the decimal point Unlocked cells can be cleared using the C key see also option key below The user mode starts with a blank grid Sudoku Setup Sudoku Game 1 Mode Computer Game Options Press KKM to display the options menu Return to the set up screen The instrument solves the game for you Clear all entries Save the game Use the left and right arrows to select a folder to store the game in Favourites Methods 1 9 press the down arrow and enter name do E T Press Cancel Q to return to the Utilities folder Version 1 0 Page 57 ACCESSORIES INSTALLATION Printer installation Version 1 0 Page 58 1 REMOVE THE POWER CABLE FROM THE INSTRUMENT Turn the instrument over and remove cap head screws from positions A and B using the Allen wrench provided 2 Turn the instrument back over and lift the accessory cover vertically upwards to remove Remove the tie wrap from the cable 3 Plug the accessory cable into the printer there is a lug to show the correct way round 4 Lower the printer onto the locating pins and push down firmly Turn the instrument over and replace the cap head screws as per step 1 Printer Switch the instrument on and go to utilities in
63. ss Y to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen n 04 0 6 08 Lo l2 14 Version 1 0 Page 48 Calibration Manual entry Shows previously entered calibration values and allows values to Standards be entered via the keypad 0 20 0 0444 15 The highlighted box can be edited in order to enter an 0 40 0 148 4 absorbance value corresponding to a given concentration value 0 60 0 249 A using the keypad numbers Range 0 001 to 9999 Use C to 0 80 0 349 A E backspace and clear the last digit entered and the up and down 1 00 04514 E arrows to move between boxes 1 40 0 542 A Biuret Calibration 4 Press OK 07 to accept the calibration and go to the Results E screen see below n 04 DE 0 8 10 Le 14 OR Press Back Q to return to the Standards screen Results screen Biuret Step 14 seenak Insert the reference and press the 0A 100 T key This will be TT used for all subsequent samples until changed Step 15 pecans Insert the sample and press O 0 249 A The concentration of the sample is taken and displayed Repeat step 15 for all samples Press Q to return to the Protein Folder Curve Fit Regression Press IIM to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above EP Farameters 2 Print result via selected method E Pm 3 Toggle
64. strument preferences and select the Built Auto Print in printer Printer Version 1 0 Page 59 Loading changing the printer paper 1 Lift off the paper cover Lock the platen and turn the knob to feed the paper 2 Feed in the paper Sometimes it helps if the platen lock is released 3 Paper gripped 4 Replace paper cover Version 1 0 Page 60 Bluetooth accessory installation 1 REMOVE THE POWER CABLE FROM THE INSTRUMENT Turn the instrument over and remove the cap head screws from positions A and B using the Allen wrench provided 2 Turn the instrument back over and lift the accessory cover vertically upwards to remove Remove the tie wrap from the cable 3 Plug the accessory cable into the Bluetooth module 5 Note the slots in the base of the case The two lugs on the Bluetooth module plug into these Version 1 0 Page 61 Version 1 0 Page 62 5 Note the slots in the accessory cover designed to engage with the Bluetooth accessory PCB 6 Lower the accessory cover vertically downwards onto the instrument engaging the PCB in the slots 7 Invert the instrument and replace the cap head screws at A and B 8 Attach the license label as shown Printer 9 Switch the instrument on and go to the preferences page under Auto Print utilities instrument and select the Bluetooth option Printer A Computer Bluetooth Version 1 0 Page 63 PRINT VIA COMPUTER
65. tion spectrum can be obtained from your instrument enabling simple identification of peak height and position The procedure is as follows Wavescan Parameters Start wavelength End wavelength 500 nm Mode et OF Wav escan 420 440 460 480 A 450mm Wav escan Height 0 3214 width 24 0nm 420 440 460 480 a ar pas pe abs 01150 0 308 0 088 014 Sample 1 a 446nm Abs 0 348 4 Version 1 0 Step 1 Set start wavelength by using keypad numbers or left and right arrows Press the down arrow key Step 2 Set end wavelength by using keypad numbers or left and right arrows Press the down arrow key Step 3 Select the mode Absorbance or T using the left and right arrows Step 4 To enter the measurements screen with the selected parameters press OK OR Cancel the selections and return to the Applications Folder by pressing Cancel O Step 5 Insert the reference Press 0A 100 key This will be used for all subsequent samples until changed Step 6 Insert sample and press Repeat step 6 for all samples Results A graph of the wavescan is displayed along with a table of Absorbance T at each peak Use the left and right arrows to move the cursor along the graph When it reaches a peak the peak height and width of the peak is displayed at the top of the screen To zoom in on the wavelength scale use the up arrow This auto scales on the Absorbance T scale dependent on t
66. tions menu Both are accessible from the home folders page Favorites This folder enables the user to quickly select any frequently used Methods Up to 9 Methods may be stored in the folder of eG eb Methods 1 Methods 2 Methods 3 of eQ ea Methods 4 Methods E Methods E of eG ef Methods Y Methods E Methods 3 Methods These are further storage folders enclosed in the top level Methods folder Up to 9 Methods may be stored in each folder Operation is identical to the Favorites Folder Saved methods can be locked unlocked and deleted using the Options menu Select the method by pressing the relevant key pad number and then press the KKA key Delete Method Press 1 to select delete method Select the method to be deleted using the left and right arrows Press lt gt to delete the method OR Y cancel to return to Favorites Methods folder Lock Method o Press 2 to select lock method SO Select the method to be locked using the left and right arrows Press the down arrow Select a pass key using the keypad numbers or left and right arrows were Press Y to lock the method o PA OR cancel to return to the Favorites Methods folder Unlock Method Unlock Method Press 3 to select unlock method Select the method to be unlocked using the left and right arrows Press the down arrow Enter the pass key using the keypad numbers or left and right arrows Press to unlock the method OR cancel to
67. ts Background color yellow symbol and outline black Unpacking Positioning and Installation e Check the contents of the pack against the packing list If any shortages are discovered inform your supplier immediately e Inspect the instrument for any signs of damage caused in transit If any damage is discovered inform your supplier immediately e Ensure your proposed installation site conforms to the environmental conditions for safe operation Indoor use only Temperature range 5 C to 35 C Note that if you use the instrument in a room subjected to extremes of temperature change during the day it may be necessary to recalibrate by switching off and then on again once thermal equilibrium has been established 2 3 hours Maximum relative humidity of 80 up to 31 C decreasing linearly to 50 at 40 C e The instrument must be placed on a stable level bench or table that can take its weight lt 4 5 kg so that air can circulate freely around the instrument e This equipment must be connected to the power supply with the power cord supplied It can be used on 90 240 V 50 60 Hz supplies e If the instrument has just been unpacked or has been stored in a cold environment it should be allowed to come to thermal equilibrium for 2 3 hours in the laboratory before switching This will prevent calibration failure as a result of internal condensation e Switch on the instrument via the keypad after it has been plugged in The instrume
68. tted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 When all standards are measured the OK box appears Press Lowry Calibration to accept the calibration an go to the Results screen see va below 0 20 OR nag Press Back to cancel selections and return to the Standards screen 0 80 a Calibration Screen replicates on iis This shows the calibration values and allows standards to be measured D2 0 4 0 56 0 8 Jo La 1 4 Step 11 Insert the reference sample Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Press Y to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press 0 to measure the standard and store the result Lowry Calibration Repeat for all replicates and standards A graph will display the results and the fitted curve as the measurements are input Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 gece me cee R A Press to accept the calibration and go to the Results screen see below OR Press Back Q to return to the Standards screen Version 1 0 Page 45 Lowry Calibration Standards olo o Ta me h Saja O 5 4 4 4 4 m S aja 0 01
69. value is 33 e range is 0 01 to 9999 OR Step 6 units pmol ul ME e Enter the proportions of bases present using the keypad ee numbers and up and down arrows to move between boxes Default is 10 for each range is 0 to 9999 Background Step 7 Press OK O to enter the Results screen OR Cancel Q to return to the Nucleic Acids folder Oligo Parameters 1mm Version 1 0 Page 33 OOo 000 Version 1 0 Parameterz Print Graph Sample Mumber Save Method Auto Print Results Screen Step 8 Insert the reference Press 0A 100 T Key This will be used for all subsequent samples until changed Step 9 Insert sample and press 0d This measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 absorbencies are calculated both corrected by the background wavelength value if selected Gives concentration based on absorbance at wavelength 1 Repeat step 9 for all samples Press Q to return to the Nucleic acid folder Press II to display available Options which are described below Options select using key pad numbers 1 Return to parameters screen step 1 above 2 Print result via selected method 3 Toggle graph on off The graph shows a wavescan plot across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if background correction selected 320 nm 7 Sample number add a prefix to the sample number and reset the incrementing
70. viously stored results before measuring Press to measure the standard and store the result 3 Repeat step 12 for all standards A graph will display the results Mene nd a and the fitted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 When all standards are measured the OK box appears Press to accept the calibration an go to the Results screen see below OR Press Back Q to cancel selections and return to the Standards screen Biuret Calibration Calibration Screen replicates on This shows the calibration values and allows standards to be measured Step 11 Sen Insertthe reference Press 0A 100 key This will be used for all subsequent samples until changed Step 12 Press Y to display the replicate entry boxes Use C to clear previously stored results before measuring Insert the standard and press 0 to measure the standard and store the result TE E eat Repeat for all replicates and standards Standards A graph will display the results and the fitted curve as the oo44 08 measurements are input 0148 a Use the up and down arrows to select a standard to be repeated 0 250 A if a poor reading has been obtained Use C to clear the previous 0 3494 m3 reading 0 446 4 0 542 A i Step 13 D L 2 9 Je A ol mu mun 4 4 4 4 4 mo oe Oo oa oa a O Pre

Hoefer Vision Life Spectrophotometer User Manual

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