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Evolution 60 User Guide - Fisher UK Extranet

1. Press ft or 1 t ee a Unit With the parameter e g AutoPrint highlighted press Enter The value will toggle to the opposite value With the parameter e g Test Name highlighted press Enter The Name Entry screen is displayed Highlight the desired character and press Add Character When completed press Accept Name reate Test Name 17 25 28Feb07 ABCDEFGH I JKLMNOPQRS TUVWXYZ0123456789 abcdefghi jk lmnopqrstuvuxyz _ p Ot Test Name Copper Test 1 Press fT or to select character Press Accept Name to save the test Name Character Character Name Utility is used to set certain non test hardware parameters such as the date and time standby setting screen contrast settings and printer setup It is also used to access a directory of all stored tests and the calculator function Evolution 60 User s Guide 21 Setting the date and time 22 To set the date Evolution 60 User s Guide You can set up the other utility parameters or change the utility settings at any time except when an entry screen is displayed or when the instrument is carrying out a measurement e Press Utility tility 11 04 28Feb07 Software Revision 0 014b Follow these steps to access the date and time settings Highlight Date Time Setup and press Enter Utility 3 39pm 16Mar07 Time Format Set Time 3 39pm Press T or to select item to change The screen displays the three date t
2. 82 To use all the data to calculate the reaction rate 82 To select specific start and end times for the rate Calculation a2 E E E AE 83 Scanning ssssssssssssssnnnnnrinrrsssssessesssnnnnnnnnnnnnnnnnnnnnnnenenennnnnnnnnnnnnnnnnnnnnnneneeeeennnna 85 Recalling a t st reniri ana n he a 85 Setting Up test parameters sssssssssrrsnrrnrrrrernrrnres 86 Collecting a baseline SCAN ssssesnrresnrrrrrrrrrrrrnrrens 87 Scanning an UnNkNOWN sssssssrrsnrrernrrrrnrrrrrrrrrrrrnrrene 88 Viewing and manipulating scan data cccceeeeee eee 88 Rescaling graphical Scan data cccceeeeeeeeeee ee ee ees 89 Performing calculations on the scan data 92 Viewing and rescaling tabular scan data 085 96 3 Point Net iisessc rian ea die ian 97 Recalling a test iiciin eeii o besxedeeebvidies 98 Setting Up test parameters sssssssrsnrenrrnrrrrernrrnrns 98 Taking measurementsS ssssssssrrssssrrrrrrsssrrrrererrrrnne 98 Multiwavelength sssssssssssseseesessssssunnnnnnnnnnnnnnnnnneennnnnnnnnnnnnnnnnnnnnnnnne 101 Recalling a testennou aae e A E a A 101 Setting Up test parameters ssssessssrrsrrrnrrrnrenrens 102 Adding wavelengths and factors ssssssessrrssrrn 102 Deleting wavelengths and factors cccceeeeeee eee 103 Taking M ASUrEMeENKS ccccceesceeeeeeeeesseeeenseeeenaaes 103 Performance Verification cccccssssssesessesssseseeseesesseees
3. A A MV A A D fs 183 f4 75 8 A A dil vol 0 Ratio _ smp vol 1 A Ave DNA ie diluent vol sample volume Start wavelength ug mL 260 280 Dilution Factor Di 225 nm with Scan sample volume Stop wavelength 325 nm DNA concentration A Arg If D A 260 nm A2 280 nm Protein concentration A A ef ir optional Aref 320 nm A Ale optional Ratio _ f 50 2 A f2 1 dil vol 0 smp vol 0 Thermo Scientific Evolution 60 User s Guide 171 Test Name Calculation s Default Displayed Parameters Results DNA are diluent vol sample volume Ai 260 nm ug mL 260 230 Dilution Factor D4 A E TA A gt 230 nm with scan P Aref 320 nm optional DNA concentration A hg M JA A D f 49 1 f2 3 48 Protein concentration A Ay MV A A D fa 183 A A y dil vol 0 Ratio _ smp vol 1 A mer dsDNA Conc F x Ass D 260 nm g mL Factor pna 50 ssDNA RNA Conc F x Ajo D 260 nm ug mL Factor spya or pya 40 Oligos Conc F x As D 260 nm ug mL entered factor Factor tigos 33 Oligos Conc F x Ao D 260 nm ug mL calc factor F factor calculated by Oligo Calculator Bradford Second order 595 nm g mL standard Std Conc of 0 200 400 600 800 1000 Bradford Second order 595 nm g mL micro Std Conc of 0 20 40 60 80 100
4. Accept Name Accepts the displayed Name entry Test Name and Edit Units Add Character Adds highlighted character to Name entry Test Name and Edit Units Add nm Allows you to add a wavelength and factor to the list in Multiwavelength tests and some Performance Verification tests Area Enables you to calculate area under peak in graph Scanning AutoPrint Turns automatic printout on or off Autoscale Rescales graph to original ranges of x and y axes Kinetics Scanning Base Sequence Sequence of bases contained in the sample Oligos calc factor tests Baseline Expiration Enters time when baseline for scan tests will need to be collected again Utility Thermo Scientific Evolution 60 User s Guide 159 Parameter Description Beeper Turns audible signal for key presses on and off Utility Calculation Baseline Select the zero baseline or the tangential baseline to calculate area under the peak in graph Scanning Calculator Enables the calculator mode Utility Cell Correction Select option to automatically corrects for variances in absorption between the cuvettes all test types Cell Position Displays position placed in light path only with auto turret Change Mode Change to Abs Change to T Switches measurement modes Basic A T C and some Performance Verification tests Collect Baseline Starts the collection
5. C d Measure Baseline Tabular Graph Sample Within the Scanning program you can view and manipulate your results either in graphical form or in tabular form When you are working with graphical scan data press Edit Graph before you perform other functions on the scan data When the edit graph screen appears you can Thermo Scientific Rescaling graphical Thermo Scientific scan data e Rescale the graph e Determine 3 point net values e Calculate area under a curve e Label peaks and valleys Scanning 13 30 20Jun02 Test Name Filter BLANK 0 730 Wavelength ID 1 Baseline collected 20Jun02 cal a el You can modify the scale of your scan data plot in two ways automatically or manually When you select Auto Scale the instrument automatically scales the X and Y axes so all the data appears on the plot When you select Manual Scale you select specific minimum and maximum values for the X and Y axes Whenever you modify the scale the instrument also automatically recalculates and displays the new data plot Press Edit Scale to modify the scale When the edit scale screen appears you can e Use the Auto Scale function to change the scale and display the new graph e Use the Manual Scale function to change the scale and display the new graph e Use the cursor to identify specific points along the X axis Evolution 60 User s Guide 89 90 To use the Auto Scale function E
6. Delay Time mi onc of Standa Sample Positio ore parameter Press fT or to select Press ENTER Press ft or to select item to change di Unit Evolution 60 User s Guide 37 2 Highlight the unit you want to select and press Enter Creating custom In addition to the basic concentration units you can units create a custom concentration unit and add it to the list This custom concentration unit can be changed when desired Note Only one custom concentration unit is available in the list at one time a 1 With the Units Selection window displayed press Edit Unit Use this screen to e Delete the name of a unit e Delete a character in the name of a unit e Add a character to the name of a unit e Accept the name of a unit Edit Unit 10 13 14Feb02 IBCDEFGH I JELMNOPQRS TUVWX YZ0 123456789 abcdefghijklmnopqrstuyvuxyz _ pOH Unit Press ft or to select character Character Name 2 Follow steps 2 through 4 in the Naming a Test section above The name of the new custom unit appears on the list of basic units 38 Evolution 60 User s Guide Thermo Scientific Cell Correction Every test setup screen provides access to the cell correction program Note The Cell Correction program is not active from the Main Basic ATC screen a Note The Cell Correction feature is active only when the 6 Position Cell Holder is set to either Auto 6 or Auto 3 The feature is
7. Na C RNA RNA hybrids units G units C 1 0 7 Na 0 7 GC f 500 L P 0 35 formamide M molarity of cation Fraction GC fraction of G and C form formamide in the sample L of base pairs P mismatching Conversion from ug MI and molecular weight from Val ug mL x 1000 pmol L i pmol L Oligo calc factor test DNA Mol Wt Thermo Scientific Evolution 60 User s Guide 175
8. Samples a Measuring unknowns i Absorbance Difference 13 29 14Feb02 Measuring unknowns Test Name TEST 9 BLANK automatically S Abs Abs Abs Result using Auto 6 or Auto 3 ID 429 0 630 0 Ref WL ppm 74 Insert samples 1 5 into positions 1 5 Press ENTER to measure 1 With the Absorbance Difference screen displayed press Run Test Evolution 60 User s Guide Thermo Scientific Measuring unknowns manually Thermo Scientific using Manual 6 or Single Cell Holder pm m Place the blank and unknowns in the correct cell positions Press Enter to start the measurement The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen With the Absorbance Difference screen displayed press Run Test Place the blank and unknowns in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder a Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position a Absorbance Difference 13 39 14Feb02 Test Name TEST 9 Cell 1 Abs Abs Abs Result ID 429 0 630 0 Ref WL ppm E SS SS Ss Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the position butto
9. Abs Ratio Abs Diff Multiwavelength tests Factor 3 31 Enters factor to convert a datum to a result Abs WL3 31 x Factor Result Multiwavelength tests Graph Displays graph of collected data Kinetics Scanning ID Enter numeric identifier for measurement autoincrements during test until turned off set to 0 Instrument Serial Number Displays serial of the instrument Utility Intercept Enter where line crosses the y axis Abs where conc 0 Interval Enter wavelength range between data points Scanning tests only Interval Time Enter time between repeated readings Kinetics only Linearity Value Enters a linearity value Kinetics only To help determine the linearity of the reaction during the measurement the spectrophotometer offers a linearity parameter This parameter is the difference between the changes in the absorbance of two measurements as shown in the following example Time Abs AA Linearity 1 1 2 2 1 3 29 09 P 4 38 09 P 5 46 08 P 6 52 06 F Linearity is the AA between AA calculations P Pass and F Fail 162 Evolution 60 User s Guide Thermo Scientific Parameter Description Load Test Loads highlighted test from Stored Tests Directory into active memory and sets instrument to test parameters Utility Lock Unlock Used to protect stored tests from accidental deletion or alteration asks fo
10. DNA 260 280 9 05 28Mar0Z Test Name DNA 260 280 Cell 1 Abs Abs IDH 260nm 280nm 1 0 460 0 243 DNA DNA Protein Ratio Conc Conc Result 1 894 20 19 28 61 Page 1 Sample 1 Sample Press Run Test Place the blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually The DNA measurement with scan tests include two tests that function almost identically the only difference is in the wavelengths used for the measurements In both cases the scan is measured from 225 to 325 nm One test measures absorbance at 260 and 280 nm while the other measures absorbance at 260 and 230 nm The measurement of DNA purity using the 260 230 ratio is commonly employed when Phenol is used in the process of extracting DNA Phenol has a high absorbance at 230 nm and is easily detected using this test method Evolution 60 User s Guide 129 Note Note Setting up test parameters Measuring Protein Concentration 1
11. Madison WI 53711 4495 U S A Telephone 1 800 642 6538 U S A or 1 608 273 5017 worldwide Fax 1 608 273 5045 worldwide E mail techsupport analyze thermo com www thermo com Trademark Information Evolution is a trademark of Thermo Fisher Scientific Inc Microsoft Windows Windows NT and Excel are either trademarks or registered trademarks of Microsoft Corporation in the United States and or other countries Adobe and Acrobat are trademarks of Adobe Systems Incorporated 269 211300 Thermo Scientific Contents ntroduction ee 1 Conventions used in this manual ssssssssssssssrrrrssssrrrss 1 Spectrophotometer BASICS sssssssssssessssrrrrresssrnnrrrsrssnnnnreresnnns 2 Spectrophotometer COMponentS sssssrsrrrrerrerernrne 2 GOMPECEOIS onise sees k tee ecctteoren AAAA A ATREA 3 About the keypad sssssssssssrsssssrrrrrrssnnnrrrssrnrrrreens 4 Cell Holders ninaongea ea a a i 6 6 Position Cell Holder ssssssssnrrnnrrrnrrrnnrrennrrernrrren 6 Sifigle Cel Holder asrnane man Aa 6 Selecting and positioning cuvettes sssssresrrrrssree 7 Z dimensSionS sa ssssrnnnnrnrnrrnrrrrnnrernnrnnnnrnnnrrnrerrnnne 8 Cell holder configurations cccccceeeseeeeesseeeeneseenees 8 ACCESSOMIES a 11 Cell holders and cell holder accessories cceeeeee ees 11 Cell holder initialization cccc cc ccccee eee eeeeeeeeeeeees 11 Changing cell holders sssssssssssrssssrssnsss
12. Refer to the Parameters chapter for a description of the parameters and Calculations for Bio Tests Software chapter for the default values To get started highlight Protein Tests and press Enter Then highlight Direct UV 280 and press Enter The Direct UV 280 parameter screen appears Note The following screens show the parameters for the Direct UV tests at 280 nm For the Direct UV test at 205 nm the wavelength is set to 205 nm a Note If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Direct UV 280 14 31 eae Sample Positioner Auto 6 umber of Samples 1 More parameters Press T or to select item to change aaa a Test Tests Test 142 Evolution 60 User s Guide Thermo Scientific Setting up test parameters Measuring the sample Measuring standards automatically using Auto 6 or Auto 3 Measuring standards manually Thermo Scientific using Manual 6 or Single Cell Holder Highlight the name of the parameter you want to set When the parameters are set press Save Test to save the test or Run Test to measure the blank and unknowns Place the blank and unknowns in the correct cell positions Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the absorbance and concentration of each unknown Press Run Test Place t
13. a E A E aanpohenmesouaniesanamah venta csesowa 31 Loading test Tesi is piren reni k cuss fea nyien piani ba iaaa 31 LockK UnlOCKa a a Speer renner 32 Delete TeStts ss scecesns ie nairean AiE EEES EE EENT 33 Sea ES as oss asa eee eee a eee 34 Setting up a single test SmartStart ccccccee cece eas 34 Setting up a multiple test SmartStart ccceeeeeaes 36 Concentration UNIS css sssessessessesessessesseseesessessesesseseeaeeaeseess 37 Specifying Concentration UNItS cccceeeeeeeeeeeee eee eees 37 Toselect the Units ces centre meer ations eta o eros 37 Creating custom UN ItS ssssssssssrsssrrresrrrerrrrnrrrnnrne 38 Cell Correction ccccccscssssssssssscssssssssssessesessessesacsssessssacsacsessesseaceaceaees 39 Running the Cell Correction Program scccesseeeeeeeees 40 Specifying wavelengths for Discrete nms mode 42 Sample Positioner Setting c ccccessseceeeesseesseeeeesteesteeeeeteens 44 UOA EATA E A ee Daca nteed 44 OEE E E eae E E E T eben 44 Single Cell Holde si mernin ian nan aaa y aa 44 Manual O ren e a e a aaa meee 45 Basic ATC Mode Abs amp T Measurements 46 Setting the wavelength ccsccecescceeeseeeenseeeeenaees 47 Measuring a Dian midisin a a a A 47 Measuring UNKNOWNS sssssssssssssssssrrrrsssrrrrrssssrrrresesnns 48 Basic Concentration measurements cceeeeeeeeeeeees 48 Setting the wavelength and Mmode sssssssssrserrrsrrrns
14. right cursor lines Select the wavelengths with Cursor and Cursor Repeat until all three wavelengths have been selected Evolution 60 User s Guide 93 94 Calculating the area under a curve Evolution 60 User s Guide 5 L 2 Press Enter Factor to access the set factor box Enter the desired factor and press Enter The instrument calculates the value for the 3 point net absorbance for the selected wavelengths multiplied by the selected factor With your scan data displayed on the edit graph screen press Math Press Area The Area Under the Curve measurement screen appears Scanning 13 50 20Jun02 Test Name Filter BLANK 440 460 0 075 0 060 0 730 A 0 000 425 Wavelength 500 Area Tangent 4 933 actor 1 000 e Nex Options Cursor paur Cursor Use Cursor and Cursor to position the left cursor line to the desired wavelength value The instrument calculates the area under the curve for the selected wavelengths Continue selecting the other wavelengths by pressing Next Cursor to activate the next cursor line Select the wavelength with Cursor and Cursor lt Press Set Options to access the set options window Thermo Scientific Labeling peaks and valleys Thermo Scientific L Highlight Factor Enter the desired factor and press Enter Highlight Calculation baseline Press Enter to toggle between Zero and Tangent Press Esc to return to the
15. 59 Standard Curve When you use the Standard Curve program you can perform these tasks e Recall a standard curve or create a standard curve set up the parameters and then measure the standards for the curve e Run the Cell Correction program e Measure unknowns e View the data select between graphical and tabular displays e Edit a standard curve change the number of standards select a different curve fit or delete points from the curve To get started press Test When the Test Types screen appears highlight Standard Curve and press Enter Standard Curve 15 28 4Feb02 BLANK Test Name AMMONIA Date Standards Measured ave length 660 Onm Ref Wavelength Correction orf urve Fit Linear umber of Standards 5 nits ppm Sample Positioner Auto 6 umber of Samples 1 ore parameters Press ft or to select item to change Pe ea e N Standards Test Tests Thermo Scientific Evolution 60 User s Guide 60 Recalling a standard curve Setting the parameters for a standard curve Measuring the standards for a standard curve Measuring standards automatically using Auto 6 or Auto 3 Thermo Scientific L Press Stored Tests Highlight the name of the test you want to recall and press Enter Place the standards in the correct cell positions Set the parameters for measuring the standards See the Parameters chapter for a complete list a
16. AT U GC sequence entered anak of G C bases x 100 Percentage Yo above total of AT or U GC Molecular weight units A units T If entry does not include U Molecular units G units C MW 312 2 x A 303 2 x T weight units U 329 2 x G 289 2 x C 18 02 x Da M If entry does include U MW 329 2 x A 306 2 x U 345 2 x G 305 2 x C 18 02 Absorptivity units A units T If entry does not include U Extinction 260 units G units C Eng 15 200 x A 8 400 x T coefficient units U Mitcm 12 010 x G 7 050 x C If entry does include U E60 15 200 x A 9 900 x U 12 010 x G 7 050 x C Conversion Factor N A Molecular Weight x 10 g mL Extinction Coefficient Calculation of Tm units A units T Tm 2 A T G C C Oligos up to 20 units G units C bases in length Calculation of Tm units A units T Tm 81 5C 16 6log Na Thermo Scientific Evolution 60 User s Guide 174 Calculation Entry Parameters Formula Displayed Units Calculation of Tm units A units T Tm 67 C 16 6log Na C DNA RNA hybrids units G units C 1 0 7 Na 0 8 GC i 500 L P 0 5 formamide M molarity of cation Fraction GC fraction of G and C form formamide in the sample L of base pairs P mismatching Calculation of Tm units A units T Tm 78 C 16 6log
17. Lowry Second order 550 nm g mL standard Std Conc of 0 100 200 500 1000 2000 Lowry Second order 750 nm g mL micro Std Conc of 0 20 50 100 200 500 BCA Second order 562 nm g mL standard Std Conc of 0 0 2 0 4 0 6 0 8 1 0 172 Evolution 60 User s Guide Thermo Scientific Test Name Calculation s Default Displayed Parameters Results BCA Second order 562 nm g mL micro Std Conc of 0 0 5 1 2 5 10 Biuret First order through zero 540 nm mg mL Std Conc of 0 2 4 6 8 10 Direct UV Conc F x Assy D 280 nm mg mL 280 Factory 1 Direct UV Conc F x As D 260 nm mg mL 205 Factory 31 Warburg Christian diluent vol sample volume A 280 nm mg mL Dilution Factor Dj A gt 260 nm sample volume fi 1 55 f2 0 76 Protein concentration A Vf A Vf pD i Cell growth None 600 nm Abs Thermo Scientific Evolution 60 User s Guide 173 Calculations for Oligo Calculator DNA DNA hybrids units G units C M molarity of cation Fraction GC fraction of G and C form formamide in the sample L of base pairs P mismatching 1 0 7 Na 0 51 GC 500 L P 0 63 formamide Calculation Entry Parameters Formula Displayed Units of bases Repetitive sequence of A T or U Count total of bases entered Length G and C of bases GC content Use
18. a 6 Position Cell Holder is installed press the cell position buttons to reposition the cell holder and measure the rest of the unknowns manually 100 Evolution 60 User s Guide Thermo Scientific Recalling a test Thermo Scientific Multiwavelength When you use the Multiwavelength program you can perform these tasks Recall a test or set up new test parameters Run the Cell Correction program Add or delete wavelengths and factors Measure a blank Measure unknowns To get started press Test When the Test Types screen appears highlight Multiwavelength and press Enter ultiwave length 10 30 22Feb07 BLANK easurement Mode Absorbance ample Positioner Auto 6 umber of Samples 1 ell Correction off ID O OFF 1 Press ft or to select item to change L 2 Set nms Test ored Tests Press Stored Tests Highlight the name of the test you want to recall and press Enter The parameters for the selected test appear on the screen From this screen you can e Set up test parameters Evolution 60 User s Guide 101 e Run the Cell Correction program e Save atest e View the list of stored tests e Measure a blank e Measure unknowns Setting up test 1 With the Multiwavelength screen displayed parameters highlight the name of the parameter you want to set Refer to the procedures below for specific instructions on adding or deleting wavelengths and factors If you have previously selected t
19. access Run Test or Measure Samples a DNA 260 280 Ref Wavelength Correction DNA Factor 260 Onm 280 Onm P 280 Onm Protein Factor 260 0nm 0 000 More parameters Press T or to select item to change ave red RuN Test Thermo Scientific Evolution 60 User s Guide 127 Setting up test parameters Measuring Protein Concentration Measuring unknowns Measuring unknowns automatically using Auto 6 or Auto 3 128 Evolution 60 User s Guide With the DNA 260 280 or DNA 260 230 screen displayed highlight the name of the parameter you want to set To measure the protein concentration using this test method select Measure Protein and press Enter When the parameters are set press Save Test to save the test under a new name or Run Test to measure the blank and unknowns DNA 260 280 Test Name DNA 260 280 Ab Abs s IDH 260nm 280nm 8 09 28Mar02 BLANK Insert sample 1 into position 1 Press ENTER to measure G es eee ee 1 Place the blank and unknowns in the correct cell positions 2 Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen Thermo Scientific Measuring unknowns manually using Manual 6 or Single Cell Holder DNA with Scan 260 280 and DNA with Scan 260 230 Thermo Scientific
20. be displayed in Utility Test Directory screen available in all tests Tm value Calculates melting temperature Oligos tests Total Run Time Enter time from Run initiation to end of test Delay Time Interval Times Measurement Times Kinetics Units Select or create units labels for results all stored tests except Abs Ratio Scanning Cell Growth Unselect Test Remove gt tag of highlighted test name to remove test from the SmartStart menu Utility Stored Tests Directory Wavelength 166 Evolution 60 User s Guide Enter values for the analytical wavelengths Thermo Scientific Calculations for Software Calculation Calculation s Graphs Standard Curves Partial sums SX x SY y SXX Sx SYY y SXY x y SOX Z x x N SXX SX SOY Zly y N sYY SY SSXY x x y y N SXY SX SY where x concentration of i standard y absorbance of i standard N number of standards Linear A A c regression where A absorbance general case c concentration A c is defined by an equation of the form A C a c a c a c 4c a where Y axis intercept a ad coefficients The coefficients are computed using the least squares method Thermo Scientific Evolution 60 User s Guide 167 Calculation Calculation s Graphs Linear regression through zero A a c where A _ absorbance c c
21. before removing the cell changer a To e use long pathlength cells cylindrical or rectangular e use test tubes e measure solid samples in the filter holder e regulate sample temperature via an external liquid recirculator you must install the appropriate cell holders The 6 Position Cell Holder installed in the spectrophotometer can easily be removed to install other accessory cell holders See Removing the 6 Position Cell Holder and the Single Cell Holder Thermo Scientific Installing the 6 Position Follow these steps to properly install the 6 Position Cell Cell Holder and the Holder or the Single Cell Holder Single Cell Holder 1 Open the sample compartment door and let it rest on its hinge 2 With one hand carefully lower the cell holder straight down into the sample compartment 6 Position Cell Holder Thumbscrew Thermo Scientific Evolution 60 User s Guide 13 Single Cell Holder Captive thumbscrews Single Alignment pin hole PAs NS 3 With the other hand tighten the captive thumbscrew s 4 Close the sample compartment door Note Ifthe cell holder is not aligned correctly you will not be able to tighten the thumbscrews a Removing the 6 Position Follow these steps to remove the 6 Position Cell Holder Cell Holder and the Single or the Single Cell Holder Cell Holder 1 Open the sample compartment door and let it rest on its hinge 2 With one hand loosen the captive
22. but you should change them to the values on the calibration certificate included with your standards You can display the tolerances for this test in either absorbance or Transmittance a When running the Photometric Accuracy test remember that e You will need Wavelength Accuracy standards that are designed to measure the wavelength accuracy at the specified wavelengths e An empty cell holder should be used as the blank e The standards need to be measured in the order that they are listed on the test screen e 1 5 standards can be used 1 Highlight Photometric Accuracy 2 Press Enter erformance Verification 16 46 2Mar07 hotometric Accuracy ft nm Expected Tol Meas 7 z T zT Result 1 590 0 50 000 1 200 2590 0 10 000 1 200 Place Standards 1 2 in positions 1 2 Thermo Scientific Selecting the mode To switch between absorbance and Transmittance press Change to T or Change to ABS until the appropriate mode appears Adding standards You will need to set three values whenever you add a standard the wavelength the absorbance or Transmittance and the tolerance value 1 Thermo Scientific If you need to add a standard press Add Std and enter the wavelength value in the Entry field When the wavelength value entry is complete press Enter or Add nm to add the wavelength to the list The screen changes to prompt you to enter the absorbance or T for the wavelen
23. in the B position The instrument always uses the B position to collect the baseline a 1 Press Run Test The DNA with Scan measurement screen appears DNA with Scan 260 280 10 37 28Mar02 Test Name DNAscan 260 280 Cell 1 3 000 0 10 225 Wavelength 325 aise e o Blank 2 Place the blank in the B position 3 Press Measure Blank to collect the baseline When the instrument is finished measuring the blank the message disappears If you want to switch between tabular and graphical displays press Graph or Tabular a With the DNA with Scan 260 280 or DNA with Scan 260 230 screen displayed follow these steps to measure the sample If a 6 Position Cell Holder is installed be sure to place the unknown in cell position 1 Evolution 60 User s Guide 131 Note Note Note DNA Direct 260 dsDNA ssDNA RNA and Oligos entered factor measurements 132 Evolution 60 User s Guide The instrument always uses cell position 1 to measure the sample a 1 Press Measure Sample to measure the sample When the instrument is finished measuring the absorbance scan it displays a graph of the scan along with the sample ID DNA ratio DNA concentration and protein concentration If you want to switch between tabular and graphical displays press Graph or Tabular a You may need to use the up and down arrow keys to view all the data for the screen a DNA with Scan 260 280 10 51 28Mar02 Test Na
24. of the baseline Scanning only Concentration Sets concentration value Conc of Standard Displays the entered concentration value Adv A T C Correction Mode Select mode for cell correction Discrete nms or Scan Cursor Goes to Cursor tracking mode to view data points in graph Kinetics Scanning Cursor gt Moves cursor right or left on graph and displays data of lt Cursor each point Kinetics Scanning Curve Fit Selects type of line fit calculation Standard Curve tests Data File Name Allows entry of name for data file when AutoSave ON Date Cell Correction Displays date when cell correction data on cuvettes was last collected Date Standards Measured Displays date when standards were last measured with this instrument Standard Curve tests Date Time Setup Enters current date and time settings for the instrument Utility 160 Evolution 60 User s Guide Thermo Scientific Parameter Description Delay Time Enters time from Test Initiation to first measurement allows for sample equilibration ADV A T C and Kinetics Delete Character Deletes last character of Name entry Test Name and Edit Units Delete File Deletes test or data file from the Stored Tests Directory Utility Delete Name Deletes entire name to allow new entry Test Name and Edit Units Delete nm Removes a wavelength and factor from
25. the characters for the name 4 Press Accept Name to accept the name and return to the previous screen Thermo Scientific Saving atest Normally you would save a test after you have entered the appropriate parameter values so you would be ona screen similar to the one shown below DNA 260 280 12 23 21Feb07 BLANK 260 Onm 280 Onm Protein Factor 280 0nm Protein Factor 260 0nm lore parameters press 1 o a skoren Jo in Test Tests Test 1 Press Save Test The Create Test Name screen appears If the desired test name is not displayed enter it using the procedure in the Naming a Test section 2 If the desired name is displayed press Accept Name A window allowing you to select whether the test is included as a SmartStart test see the SmartStart section is displayed 3 Highlight the appropriate test and press Enter The test is now saved in the instrument library Loading test files Saved test files can be loaded from the internal memory from the Utility screen 1 To access all test files press Utility 2 Highlight Stored Test Directory and press Enter Thermo Scientific Evolution 60 User s Guide 31 Utility Test Directory 12 25 21Feb0 ucleic Acid Tests DNA 260 280 Read onl DNA 260 230 Read only DNA with Scan 260 280 Read only DNA with Scan 260 230 Read only dsDNA Read only ssDNA RNA Read only Oligos Centered factor Read only Oligos calc factor Re
26. the light beam An alignment mark on the test tube helps you orient the test tubes consistently and correctly a When using small aperture cells e Always used masked cells e Use the same cell or cuvette for your blank and your samples The figure below illustrates the position of the light beam in the spectrophotometer The specifications for the compartment including the dimensions of the beam are e Z dimension e Square cuvettes test tubes 8 5 mm e Beam size 2 mm wide by 7 mm high zm 5mm 8 5 mm The following table shows the cell holders and cell holder accessories available for the spectrophotometer These accessories may be installed or removed without the need to switch off the instrument Thermo Scientific Cell Changer System Single Cell System Standard Cell Holders Accessory Cell Holder Systems Single Cell Recirculator Temperature Control a S a Wo Y a 50 mm Rectangular Long Pathlength Cell Holder Must be installed in the B 2 and 4 positions Test Tube Holder Must be installed in the B 2 and 4 positions Must be installed in the B 2 and 4 positions 50 mm Cylindrical Long Pathlength Cell Holder Ds Thermo Scientific Evolution 60 User s Guide 9 Must be installed in the B 2 and 4 positions Cell Changer System Single Cell System 100 mm Rectangular Long Pathlength Cell Holder Cannot use 100 mm path cells with Cell Po
27. the list Multiwavelength and some Performance Verification tests Diluent Volume Enters volume of diluent added prior to measurement Dilution Multiplier in some Bio Tests Dilution Multiplier Displays factor used to correct for sample dilution Display Activity Indicates whether results should include protein concentration DNA 260 Calculates extinction coefficient DNA Factor Enter factor to calculate DNA concentration DNA Bio Tests Edit Allows you to change a wavelength or factor in the list Multiwavelength and some Performance Verification tests Edit Curve Allows you to manipulate graph Kinetics Edit Data Enables you to select portion of the data in a table for recalculation of a result Kinetics and Scanning Edit Graph Allows you to manipulate graph Scanning Edit Scale Allows you to change graph axis scales and view individual data points Scanning Factor Enters factor to convert a datum to a result Abs x Factor 1 Concentration Result Abs min x Factor 2 Kinetics Result Can be entered or calculated from concentration and absorbance values in ADV A T C Thermo Scientific Evolution 60 User s Guide 161 Parameter Description Factor 1 Enters factor to convert a datum to a result Abs WL1 x Factor Result Abs Ratio Abs Diff Multiwavelength tests Factor 2 Enters factor to convert a datum to a result Abs WL2 x Factor Result
28. them with new ones Ensuring your cuvettes are clean both inside and out is important to the quality of your results for two reasons 1 contaminating material may absorb light resulting in falsely high absorbance readings and 2 contaminants in the cell may react chemically with subsequent reagents or standards introduced into the cell Cleaning methods depend to some extent on the nature of the contaminating material It is important to identify the residual material in the cell that needs to be removed Refer to the following table for suggestions on cleaning methods solvents and material Examples Suggested Cleaning Methods Protein Biologics DNA Warm water with detergent e Dilute Nitric Acid lt 10 rinse e Copious water rinse Salt solutions e Dilute Nitric Acid lt 10 rinse e Copious water rinse Thermo Scientific Thermo Scientific Solvent Aqueous Organic Organic Organic Organic Notice Examples Basic solutions Hydrocarbons small molecules oils Alcohol solutions Acidic solutions Basic solutions Suggested Cleaning Methods Warm water with detergent Dilute Nitric Acid lt 10 rinse Copious water rinse Rinse with organic solvent Warm water with detergent Dilute Nitric Acid lt 10 rinse Copious water rinse Rinse with similar alcohol acetone or other solvent Copious water rinse Rinse with organic solvent Warm water with detergent Dilute Nitric A
29. thumbscrew 3 With the other hand pull straight up on the cell holder and lift it out of the sample compartment 4 Close the sample compartment door 14 Evolution 60 User s Guide Thermo Scientific Installing accessory cell holders Note Note Note Installing the internal printer Thermo Scientific optional A Caution Make sure that you have the correct cell holder baseplate installed If you want to use 100 mm long pathlength cells you must install the Single Cell Holder baseplate a See the Cell Holder Configurations section for the different cell holder accessories that can be created for your instrument Each of the Cell Holders need to be installed on either a single cell baseplate or a multi cell baseplate by removing the cell holder s secured to the baseplate Each cell holder has a captive screw in the bottom of the holder that secures the holder to the baseplate Use a flat blade screwdriver to loosen the captive screw from the baseplate and lift the cell holder from the baseplate Then insert the new cell holder into the appropriate position and secure it to the baseplate by tightening the captive screw You can install only three each of some accessory cell holders Make sure to place them in position B 2 and 4 a Follow the steps in the Installing the 6 Position Cell Holder and the Single Cell Holder section above for instructions on installing the complete accessory assembly
30. to measure a blank or unknowns I Advanced A T C 11 49 4Feb02 Taking measurements Test Name IRON TEST Cell 4 Abs IDH 540 Onm 1 0 050 2 0 022 3 0 036 4 0 076 Page 1 Samples i 4 Data Samples 54 Evolution 60 User s Guide Thermo Scientific Taking measurements automatically using Auto 6 or Auto 3 Taking measurements manually Thermo Scientific using Manual 6 or Single Cell Holder _ m Press Run Sample Place the blank and the unknowns in the correct cell positions Press Measure Sample to measure the unknowns The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen Press Run Sample The Advanced A T C measurement screen appears prompting you to place your samples in the cell holder Place the blank and unknowns in the cell holder If the 6 Position Cell Holder is installed be sure to place the blank in the B position You can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Press Measure Sample to measure the unknowns The sample measurement appears on the screen If a 6 Position Cell Holder is installed press the cell position buttons to reposition the cell holder
31. 00 Interval Time min sec 0 01 Total Run Time Chr min sec 6 05 00 easure Blank Once isplay Result Rate Factor On More parameters Press T or to select item to change hae ae ae a From this screen you can e Set up test parameters e Run the Cell Correction program e Save atest e View the list of stored tests e Measure a blank e Measure unknowns With the Kinetics screen displayed highlight the name of the parameter you want to set When parameters are set press Save Test to save the test or Run Test to measure a blank or unknown If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a If Auto Save Data is ON you must enter a Data File Name before you can access Run Test or Measure Samples a Evolution 60 User s Guide 77 Measuring unknowns 78 Evolution 60 User s Guide Note If a 6 Postion Cell Changer is installed be sure to place the blank in position B and the unknown in cell position 1 The instrument always uses cell position 1 to scan the unknown a 1 With the Kinetics test parameters displayed on the screen press Run Test 2 If a 6 Position Cell Holder is installed place the blank in position B and the unknown in position 1 3 Press Measure Sample to measure the blank and the unknown When the instrument completes the measurement the kinetics data and rate appear on the screen 4 If a Sing
32. 30 Evolution 60 User s Guide Refer to the Parameters chapter for a description of the parameters and the Calculations for Bio Tests Software chapter for the default values To get started with the Bio Tests screen displayed highlight Nucleic Acid Tests and press Enter A list of nucleic acid test appears Highlight DNA with Scan 260 280 The DNA with Scan 260 280 parameter screen appears The following screens show the parameters for the DNA 260 280 test For the DNA 260 230 test Wavelength 2 is set to 230 nm a If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a DNA with Scan 260 280 16 22 21Feb07 BLANK Onm 280 Onm 280 Onm Protein Factor 260 0nm More parameters With the DNA with Scan 260 280 or DNA with Scan 260 230 screen displayed highlight the name of the parameter you want to set To measure the protein concentration using this test method select Measure Protein and press Enter When the parameters are set press Save Test to save the test under a new name or Run Test to measure the blank and unknowns Thermo Scientific Collecting a baseline scan Note Note Measuring the sample Thermo Scientific With the DNA with Scan 260 280 or DNA with Scan 260 230 screen displayed follow these steps to collect a baseline scan If a 6 Position Cell Holder is installed be sure to place the blank
33. 49 Measuring a blank sssssssssssssrsssssrrrrrssrrrrrrsssrrrrrsssnn 49 Thermo Scientific Thermo Scientific Basic ATC Mode Concentration Measurements 50 Using Conc Std to measure concentration 666088 50 Using Conc factor to measure concentration 51 Measuring UNKNOWNS ccccecccee ee eeeeeeeeeetesaaeeeeeeenaaes 52 Advanced A T C Abs amp T oovesscsssststeseessessseneesesseneseees 53 Recalling a tests s 2sste Sei ie eae 54 Setting up test paraMeters ccceee cece eeceeeeeeeeeeeeeeees 54 Taking measurements cc ceceee eee eeeeeee ee eessaeeeeeenees 54 Advanced A T C Concentration sses 56 Recalling a testisi Geira eeevtelansehii chy peewee EEE 56 Setting up test paraMeters c cece eee e ccc eeeeeeeeeeeeeees 57 Measuring a standard ccc ee esee cece eee ee eee eeeeeaeeeeeeeees 57 Entering a faCtOMr serieren urre enr SEDE IE REE DEE EEEN 58 Measuring UNKNOWNS ssssssssssnrnssnrrrenrrerrrrrrnrrnrrernene 59 Standard CUTfVe assssssessessossnernnnrnnnnnnnunnnunnnunnnnnonunnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 60 Recalling a standard CUFV s ssssssrssnrrnsnrrnrrrrrrrrrnnra 61 Setting the parameters for a standard curve 65 61 Measuring the standards for a standard curve 61 Measuring UNKNOWNS sssssssssrrsrossnnnnressnnnnressnnnnreesa 64 Editing a standard CUrve ssssssssssresnrrrsnrrrsrrrrrrrrenra 66 To edit the concentratio
34. 9 Calculations for Software 20 cccececccecessstsesesesesesssseseseseseseeeeers 167 Calculations for Bio Tests Software cccceeeeeees 171 Calculations for Oligo Calculator 174 Thermo Scientific Conventions used in this manual Note Notice A Caution A Warning A Danger Thermo Scientific Introduction Congratulations on your purchase of a Thermo Scientific Evolution 60 spectrophotometer Our spectrophotometers integrate advanced hardware features with the power and flexibility of a wide range of accessories This manual includes safety precautions and other important information presented in the following format Notes contain helpful supplementary information a Follow instructions labeled Notice to avoid damaging the system hardware or losing data a Indicates a hazardous situation which if not avoided may result in minor or moderate injury It may also be used to alert against unsafe practices a Indicates a hazardous situation which if not avoided could result in death or serious injury a Indicates a hazardous situation which if not avoided will result in death or serious injury a Evolution 60 User s Guide 1 Spectrophotometer Basics This chapter describes the major components of your spectrophotometer Spectrophotometer Tne following illustration identifies some major components components visible on the outside of a typical Evolution 60 spectrophotometer Key O K
35. A tests but the factor used to convert absorbance to concentration is different Similarly for the Direct UV measurement of oligonucleotides tests the parameters are also the same but the factors used to convert absorbance to concentration are different For a complete list of all parameters and calculations for each test refer to Parameters or Calculations for Software Calculations for Bio Tests Software Calculations for Oligo Calculator Thermo Scientific DNA 260 280 and These tests function almost identically the only DNA 260 230 difference is the wavelengths used for the measurements One test measures absorbance at 260 and 280 nm while the other measures absorbance at 260 and 230 nm The measurement of DNA purity using the 260 230 ratio is commonly employed when Phenol is used in the process of extracting DNA Phenol has a high absorbance at 230 nm and is easily detected using this test method Refer to the Parameters chapter for a description of the parameters and the Calculations for Bio Tests Software chapter for the default values To get started with the Bio Tests screen displayed highlight Nucleic Acid Tests and press Enter A list of nucleic acid test appears Highlight DNA 260 280 Note The following screens show the parameters for the DNA 260 280 test For the DNA 260 230 test Wavelength 2 is set to 230 nm a Note If Cell Correction is ON you must run the Setup Correction program before you can
36. Abs mg L 660 Onm Page 1 Standards 1 5 a leeds Blank Test Standards 3 Enter the concentration value into the Entry concentration field 4 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 5 Press Measure Standards to measure the standards If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the standards manually When all of the standards have been measured the Standards screen appears showing the absorbance of each standard along with the slope intercept and correlation coefficient of the standard curve Thermo Scientific Evolution 60 User s Guide 63 Measuring unknowns Measuring unknowns automatically using Auto 6 or Auto 3 64 Evolution 60 User s Guide Standard Curve 15 50 4Feb02 Test Name AMMONIA Cell 5 Std Conc Abs ppm 660 Onm 1 0 2 0 200 0 140 3 0 250 0 152 4 0 400 0 268 5 0 500 0 312 urve Fit Linear lope 0 6091 Std Dev 0 011 Intercept 0 01272 Corr Coeff 0 995 Page 1 Standards 1 5 Q e Test Standards Test You can use this screen to e Display a graph of the standard curve data press View Graph e Save the standard curve press Save Test e Edit the standards press Edit Standards e
37. Enter the Test Name Wavelength Reference Wavelength Correction and Reference Wavelength b Select the Curve Fit Units and Sample Positioner c Set the Number of Standards and Number of Samples d Enter the low and high limits e Select the settings for the Statistics and AutoPrint functions f Run the Cell Correction program Place the blank and standards in the correct cell positions When all the parameters are correct press Run Standards to set up and run the standards Evolution 60 User s Guide 61 62 Measuring standards manually using Manual 6 or Single Cell Holder Evolution 60 User s Guide L Btandard Curve 15 39 4Feb02 Test Name AMMONIA BLANK Std Conc Abs Entry Enter a number 0 001 to 9999 Enter the concentration value into the Entry concentration field and press Enter Press Measure Standards to measure the blank and standards The instrument automatically measures the blank first and then measures the standards When the instrument has measured all the standards the Standards screen appears showing the absorbance of each standard along with the slope intercept and correlation coefficient of the standard curve Place the blank and standards in the correct cell positions When all the parameters are correct press Run Standards to set up and run the standards Thermo Scientific Standard Curve 12 50 14Feb02 Test Name AMMONIA BLANK Std Conc
38. Evolution 60 Local Control User s Guide SS The information in this publication is provided for reference only All information contained in this publication is believed to be correct and complete Thermo Fisher Scientific shall not be liable for errors contained herein nor for incidental or consequential damages in connection with the furnishing performance or use of this material All product specifications as well as the information contained in this publication are subject to change without notice This publication may contain or reference information and products protected by copyrights or patents and does not convey any license under our patent rights nor the rights of others We do not assume any liability arising out of any infringements of patents or other rights of third parties We make no warranty of any kind with regard to this material including but not limited to the implied warranties of merchantability and fitness for a particular purpose Customers are ultimately responsible for validation of their systems 2007 by Thermo Fisher Scientific Inc Madison Wisconsin 53711 All world rights reserved No part of this publication may be stored in a retrieval system transmitted or reproduced in any way including but not limited to photocopy photograph magnetic or other record without prior written permission For technical assistance please contact Technical Support Thermo Fisher Scientific 5225 Verona Road
39. Measure your samples press Run Test Press Run Test The Samples screen appears Place the blank and unknowns in the correct cell positions Press Enter to measure The instrument automatically measures the blank first and then measures the unknowns When the instrument has measured all the unknowns the Standard Curve screen appears showing the absorbance and concentration of each unknown If you want to switch between tabular and graphical displays press View Graph Tabular Thermo Scientific Measuring unknowns manually using Manual 6 or Single Cell Holder Thermo Scientific L 2 3 Standard Curve 16 05 4Feb02 Test Name AMMONIA BLANK p 3 0 1 ppm F urve Fit Linear lope 0 6091 Std Dev 0 011 Intercept 0 01272 Corr Coeff 0 995 m Ee Test Press Run Test Place the blank and unknowns in the correct cell positions Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When the blank has been measured it returns to its previous cell position Press Measure Samples to measure the unknowns If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually When the instrument has measured all the unknowns the Standards screen appears showing the absorbance and concentration of each
40. Min Y Max Y Min X or Max X to accept it Thermo Scientific Thermo Scientific Cursor function L The instrument redraws the plot using the minimum and maximum value you have entered and displays the recalculated rate and result Continue until you have entered all the values you want to change With your kinetics data displayed on the edit screen press Cursor The cursor options appear Kinetics 3 35pm 27Jan01 Test Name ENZYME BLANK 0 740 A 0 583 Time 3 00 Time Abs 1 30 0 644 Press Cursor or Cursor to position the cursor line on the appropriate point on the graph The instrument displays the data for the selected point When the cursor line is in the correct position press Set Min X or Set Max X to accept the selected point The instrument redraws the plot using the minimum and maximum value you have selected and displays the recalculated rate and result Evolution 60 User s Guide 381 Rescaling and when you are working with tabular kinetics results you recalculating tabular eed to press Edit Data before you can rescale and nan iculate kinetics results 12S After collecting your kinetics data you can use all the data for the rate calculation or you can select specific start and end times Whenever you modify the start and end times or select all the data the instrument automatically recalculates and displays the new reaction rate and result When the
41. Scientific The correction factor allows you to scale the readings from your Evolution 60 to match another spectrophotometer in your lab Simply measure the difference between the two instruments and use the factor to adjust for this difference For more information contact your sales representative When the parameters are set press Save Test to save the test or Run Test to measure the blank and unknowns L Place the blank and unknowns in the correct cell positions 2 Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the absorbance and concentration of each unknown L Press Run Test 2 Place the blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder 3 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually Evolution 60 User s Guide 147 Oligo calculator Using the oligo calculator 148 Evolution 60 User s Guide When the instrument has measured all the unknowns it displays th
42. Turn off and unplug the instrument from the wall outlet or power strip 2 Position the instrument so you can access the power entry module on the back of the instrument Evolution 60 User s Guide 155 3 Remove the power cord 4 Insert a flat blade screwdriver into the notch on the fuse cover and pry off the cover 5 Use a flat blade screwdriver to remove the fuse holder 6 Unsnap both fuses to remove them 156 Evolution 60 User s Guide Thermo Scientific 7 Insert the new fuses pushing them in so they snap into place 8 Replace the fuse cover 9 Replace the power cord 10 Plug the instrument back in to the wall outlet or power strip and turn on the power Note If the fuse blows again contact your distributor or technical support a Thermo Scientific Evolution 60 User s Guide 157 Parameters Parameter Description X Enters math operators when in Calculator mode Utility Formamide Enter Percentage of formamide contained in the sample Oligos tests GC Calculates Percentage of GC pairs contained in the sample Oligos tests Mismatch Enter Mismatch value to calculate Tm Oligos tests of lamp life used Displays estimated percentage of lamp life used based on typical lamp life of five years Utility 3 Pt Net Enables you to calculate peak height from tangential baseline in graph Scanning Absorbance Enters absorbance value
43. ad only Direct UY 260 Read only Protein Tests Bradford Standard Read only Bradford Micro Read only Loury Standard Read only ore tests Press ft or to select test 3 Highlight the test you want to load and press Load Test Note You can view the stored tests of a particular test type by pressing Test selecting a test type and pressing Stored Tests a Lock Unlock To lock or unlock a test highlight the name of the test you want to lock or unlock and press Lock Unlock Enter the password and press Enter Utility Test Directory 9 31 22Feb0 Protein Tests Lowry Micro Read only BCA Standard Read only BCA Micro Read only Biuret Read only Direct UV 280 Read only Direct UY 205 Read only Warburg Christian Read only Cell Growth Cell Growth Read only Scanning Assau5S1 VATA TY ore tests Press fT or to select test aes a a al Test Test Test Note If you want to lock or unlock access to the file you must enter the software password of this manual a 32 Evolution 60 User s Guide Thermo Scientific Delete Test To delete a test highlight the name of the test you want to delete and press Delete Test Thermo Scientific Evolution 60 User s Guide 33 SmartStart The SmartStart feature enables you to customize your spectrophotometer for your lab If you select one test as a SmartStart test the instrument when powered on will automatically load this test and set up the instrument for imme
44. advice The Internal Printer test allows you to verify that the internal printer is functional To run the test you will need to have an internal printer installed Running the internal printer test takes no more than 20 seconds after you press Stop 1 From the Utility screen verify that the internal printer is installed properly and is selected If necessary press Utility and then select the internal printer 2 With the Performance Verification screen displayed highlight Internal Printer Test 3 Press Enter Evolution 60 User s Guide 119 Performance Verification 16 57 2Mar07 Internal Printer Tes Press Start Test to start printer test Press Start Test to run the test The test print routine appears on the printed results While the test is running you can press Stop Test to stop it If the test fails follow these guidelines 120 Evolution 60 User s Guide Make sure that the internal printer is the selected as the printer device on the Utility screen Make sure that the internal printer is installed correctly Return to the main screen and press Enter If the paper does not move the printer may not be installed correctly Make sure that the thermal paper is threaded with the thermal side toward the printer head the outside surface of the roll is the thermal surface Contact technical support for more troubleshooting advice Thermo Scientific Thermo Scientific RS232 test The RS232C
45. all the standards the Standards screen appears showing the absorbance of each standard along with the slope intercept and correlation coefficient of the standard curve You can use this screen to Edit the standards press Edit Standards Display a graph of the standard curve data press View Graph Save the standard curve press Save Test Measure your samples press Run Test With the Standards screen displayed place the blank and unknowns in the correct cell positions 1 Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the absorbance and concentration of each unknown Press Run Test Place the blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Evolution 60 User s Guide 141 4 Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually Direct UV 280 and The Direct UV methods determine protein concentration Direct UV 205 based on absorbance at either 280 or 205 nm
46. and measure the rest of the unknowns manually Evolution 60 User s Guide 55 Advanced A T C Concentration When you use the Advanced A T C program for concentration measurements you can perform these tasks e Select the measurement mode you want to use concentration with one standard or concentration with a factor e Recall a test or set up new test parameters e Measure a standard or enter a factor only if you select either concentration with one standard or concentration with a factor e Measure a blank and unknowns To get started press Test When the Test Types screen appears highlight Advanced A T C and press Enter Advanced A T C 15 40 26Jun02 Test Name AAA easurement Mode Conc Std avelength 546nn Ref Wavelength Correction off Delay Time min sec 0 00 onc of Standard 100 0 More parameters Press T or to select item to change Recalling a test 1 Press Stored Tests 2 Highlight the name of the test you want to recall and press Enter or Load Test Thermo Scientific Evolution 60 User s Guide 56 Setting up test 1 parameters Measuring a standard Measuring astandard automatically using Auto 6 or Auto 3 Thermo Scientific The parameters for the selected test appear on the screen Highlight the name of the parameter you want to set Some parameters appear only if you select one of the concentration modes while others appear regardless of the measureme
47. anned measurements a If you want to set a baseline expiration time press Utility and then highlight Baseline Expiration Press Enter and set the desired time a To get started press Test When the Test Types screen appears highlight Scanning and press Enter L Press Stored Tests 2 Highlight the name of the test you want to recall and press Enter The parameters for the selected test appear on the screen Evolution 60 User s Guide 85 Note Setting up test parameters Evolution 60 User s Guide If you want to switch between tabular and graphical From this screen you can e Set up test parameters e Set up Cell Correction program Scanning 15 30 14Feb02 Test Name Cell 1 3 000 A 0 10 400 0 Wavelength 500 0 ID 1 2 a a ry Baseline displays press Graph Tabular a L 2 Highlight the name of the parameter you want to set When the parameters are set press Save Test to save the test or Run Test to measure a blank or an unknown 10 19 30May02 Scanning Transmittance Start Wavelength 300 Onm Stop Wavelength 800 Onnm Sample Positioner 1 Cell Platform Scan Speed Fast Interval 0 1inn ell Correction Needs Auto6 3 ID O OFF 1 Auto Print On On LE30O Thermo Scientific Note If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Note If Auto Save Data is ON
48. anually using Manual 6 or Single Cell Holder 140 Evolution 60 User s Guide L The instrument automatically measures the blank first and then measures the standards When the instrument has measured all the standards the Standards screen appears showing the absorbance of each standard along with the slope intercept and correlation coefficient of the standard curve Bradford Standard Test Name Bradford StdaA 11 15 i Std Curve Fit a0 3 364E 07 fal 6 97E 05 Page 1 Standards 1 4 1 e ol gas In Test Standards Test Linear Regr 2nd Order a2 0 09845 Std Dev 0 000 Place the blank and standards in the correct cell positions When all the standards are correct press Measure Standards to set up and run the standards Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Press Measure Standards to measure the standards If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the standards manually Thermo Scientific Measuring protein samples Measuring standards automatically using Auto 6 or Auto 3 Measuring standards manually Thermo Scientific using Manual 6 or Single Cell Holder When the instrument has measured
49. area under a curve screen The instrument calculates the area under a curve for the selected wavelengths factor and calculation method With your scan displayed on the edit graph screen press Math Scanning 13 40 20Jun02 Test Name Filter BLANK 0 730 A FATS Bers off Label Valleys orf Features 1 Press Tf or to select Press ENTER to change Press ESC to exit Wavelength 500 ID 1 eee collected 20Jun02 Press Peaks amp Valleys The Label Peaks and Valleys window appears Select the type of labels you want displayed and press Enter The instrument labels the selected items on your scan data plot Evolution 60 User s Guide 95 Note Viewing and rescaling tabular scan data To use all the scan data To select specific start and end 96 wavelengths Evolution 60 User s Guide Up to nine peaks or valleys can be calculated and displayed a When you are working with tabular scan data you need to press Edit Data before you can perform other functions on the scan data Scannin g Test Name FILTER Wave length 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 e IDH 1 Baseline collected 15Feb02 Use a d Bee Re ee With your table of scan data displayed on the edit screen press Use All Data L With your table of scan data displayed on the edit screen highlight the appropriate data point in the table 2 Press Start nm or End nm The i
50. as been saved the values for all parameters stored with that test will replace the current values for the test parameters again Software password This password allows you to lock test setups test parameters so that they may not be overwritten or deleted The password also allows you to remove the security so that you may edit the test parameters Please refer to the Lock Unlock section for more information on locking a test Note This password cannot be changed a Password 4363797 Thermo Scientific Evolution 60 User s Guide 29 30 Naming a test Evolution 60 User s Guide When you save tests you need to specify the name you want to use for the file You can enter an alphanumeric name up to 16 characters long 1 After setting up the values for the test parameters highlight Test Name and then press Enter Create Test Name 12 21 21Feb07 BCDEFGH I JKLMNOPQRS TUVWXYZ0123456789 abcdefghijklmnopqrstuyuxyz _ pOH Test Name Press f or to select character Press Accept Name to save the test eae P Character Name You can use this screen to e Delete the name of a test e Delete a character in the name of a test e Add a character to the name of a test e Accept the name of a test 2 Highlight the first character you want to use for the name of your test and press Add Character to add the selected character to the name 3 Continue selecting and adding characters until you have selected all
51. asurement This test measures the amount of noise nms at 260 nm All test parameters are determined by instrument specifications and cannot be changed by the user When running the noise test remember that e The OA measurement should be performed with the cell holder empty The 2A measurement should be performed with a 2A filter 1 Highlight Noise Measurement 2 Press Enter Per formance Validation Peak to Peak Meas Result Tolerance Abs 0 002 0 000 Pass 0 001 0 000 Pass Peak to Peak Meas Result Tolerance Abs Onm 50 003 0 000 Pass Onm 0 002 0 000 Pass ake sure the Blank position is empty Place 2A filter in Cell Position 1 Press Start Test 3 Make sure that the Blank position is empty and insert the 2A filter in position 1 The test results at 2A should be ignored if you do not have a filter installed in position 1 4 Press Start Test to run the test The results of the test appear on the screen indicating pass or fail for each wavelength If the test fails follow these guidelines Thermo Scientific Evolution 60 User s Guide 117 Stray Light 118 Evolution 60 User s Guide Repeat the test twice to verify that the test is failing consistently Make sure the instrument is warmed up for at least 30 minutes with the standby mode feature turned off Make sure the instrument is plugged into a stable power supply Contact technical support for more troubleshooting advice This te
52. asurements taken and then press Set nm again Press Change Mode until the appropriate measurement mode Concentration with Standard or Concentration with Factor appears Place the blank in the cell holder If your instrument has a 6 Position Cell Holder installed be sure to place the blank in the B position Press Measure Blank to measure the blank When the instrument is finished measuring the absorbance of the blank the message disappears Evolution 60 User s Guide 49 Using Conc Std to measure concentration Thermo Scientific Basic ATC Mode Concentration Measurements L If necessary press Change Mode to switch to the Concentration with Standard mode Conc Std 16 15 3iJan02 Factor 1 Std 1 C BLANK 0 478C 660 Onm Berl 7 eae Blank Standard Mode If your instrument has a 6 Position Cell Holder installed be sure to place the blank in the B position and the standard in position 1 2 Press Measure Blank to measure the blank If the Single Cell Holder is installed remove the blank and place the standard in the cell holder 3 Press Units Standard to set the units and measure the standard 4 Press Enter Conc then enter the concentration value of the standard and then press Enter 5 Press Select Units then highlight the appropriate unit in the list and press Enter to select the units for the concentration Evolution 60 User s Guide 50 Using Conc factor 1 to
53. ce representatives perform procedures requiring removal of the instrument cover and replacement of electrical components To protect both yourself and the instrument be sure to contact an authorized service representative to perform any service procedure you do not feel comfortable performing a Routine care for the spectrophotometer does not require a lot of time To help minimize maintenance time and to increase the life and performance of your instrument please follow these guidelines e Always replace the dust cover when the instrument is not turned on to prevent dust from accumulating in and on the instrument e Do not use or store the instrument in a corrosive environment e Gently wipe the outside of the instrument including the keypad with a soft cloth to remove any dust or Evolution 60 User s Guide 151 Cleaning and maintaining cells Solvent Aqueous Aqueous 152 Evolution 60 User s Guide spills Water isopropyl alcohol and other common laboratory cleaning agents may be used if necessary Always clean up spills as soon as they occur to prevent or minimize damage to the instrument If concentrated acids or bases or any hydrocarbon materials are spilled on the instrument be sure to clean up the affected area immediately Carefully check the condition of the cuvettes and other cells used to measure samples If they are chipped cracked or scratched it is important to discard the damaged cell s and replace
54. centration value Standard Curve test type Smoothing Turns data smoothing on and off Scanning Software Revision Displays version of firmware in instrument Utility SRE tolerance Enters the acceptable Stray radiant energy acceptable Standard Concentrations Enters concentration of standards used to generate standard curve for the test Standby Selects time since last keystroke or instrument activity powers down unit to save lamp life Utility Start wavelength Enter beginning wavelength for a scan Scanning tests only Statistics Turns stats on or off if ON calculates average and Std Dev of results Statistics registers are cleared when Statistics OFF and or when instrument is OFF and or when test parameters are changed and or when test is saved or resaved in all test types except Kinetics Scanning Multiwavelength Thermo Scientific Evolution 60 User s Guide 165 Parameter Description Std Concentration Enters concentration of analyte in standard solution Stop wavelength Enter ending wavelength for a scan Scanning tests only Stored Tests Directory Displays list of tests stored in the instrument Utility Tabular Displays list of collected data Kinetics Scanning Test Name Allows user to enter alphanumeric name max of 16 characters for test name will be included on data printout and if the test is saved will
55. cid lt 10 rinse Copious water rinse Rinse with organic solvent Warm water with detergent Dilute Nitric Acid lt 10 rinse Copious water rinse Keeping the cell clean is very important for a long cell life a Never store cuvettes long term in a water or solvent bath between uses If the solvent you are using dries impurities in the water or solvent be deposited on the inside of the cell causing permanent damage Evolution 60 User s Guide 153 154 Evolution 60 User s Guide Note Notice Notice e Use only lens cleaning tissue paper or fine soft cloth to wipe optical surfaces Most paper products such as facial tissues paper towels etc contain wood fibers that can damage the cell material e At the end of the day ensure that all cells are well cleaned and stored in a suitable container after drying Term Dilute acid Acid Solvent rinse Copious water rinse Detergent Definition Dilute Nitric Acid lt 10 Hydrochloric 5M acid or Nitric Acid 5M see Note below Rinse with the solvent that was originally used solvated your analyte Use a pure water e g deionized distilled RO and rinse at least 10 times Use a neutral pH detergent Triton X 100 if available to dilute acid wash water rinse to remove residue Do not use 5M Nitric Acid on an Anti reflection mirror coated cells a It is not recommended that you use an ultrasonic cleaning baths for your cells Each bath gene
56. ct Test to remove the test from the SmartStart menu The Main Menu will be displayed upon power up Evolution 60 User s Guide 35 Setting up a multiple 1 Follow steps 1 through 3 as outlined in Setting test SmartStart up a Single test SmartStart 4 Continue scrolling through the list and add tests until you have made all the appropriate selections An arrow sign gt will indicate the tests selected for the SmartStart menu You can now either press Esc to return to the Utility screen or you can power down the instrument To remove tests from the SmartStart menu see To Unselect a Test 36 Evolution 60 User s Guide Thermo Scientific Specifying concentration units To select the units Thermo Scientific Concentration Units When you run concentration or kinetics tests one of the test parameters is Units which labels the results The spectrophotometer includes a set of basic concentration units and you can also enter custom units if you wish All programs in the spectrophotometer use the same list of basic units C concentration mg mL ppm ug L ppb M L g L mM L mg L TU In addition to these units you can create your own custom unit using a character list like the one described in Naming a Test Once you create a custom unit it will appear in the list that you use to select the units 1 Highlight the Units parameter on the screen and press Enter 10 09 14Feb02
57. d time into the Entry baseline expiration time field and press Enter To make it easier to read the display you can adjust the screen contrast on the spectrophotometer Highlight Screen Contrast and press Enter tility 3 48am 16Mar07 Software Revision Instrument Serial Number 2M7A123005 Printer nternal Date Time Setup 3 48am 16Mar07 Standby 15 min Baseline Expiration Chr min Press fT for darker contrast Press for lighter contrast Press ESC to exit 2 Adjust the screen contrast by following the 3 26 Evolution 60 User s Guide instructions on the screen When the screen contrast is correct press Esc Thermo Scientific Setting up the Toset up the printer properly you need to load the internal printer Paper and set the utility parameters for the printer Before setting up the parameters for the printer be sure that the printer is installed If you have ordered the internal printer as a separate item you will need to install it 1 Install internal printer Refer to the Installing the Internal Printer section in the Accessories chapter for instructions on installing the printer 2 Load paper in the printer Refer to the Loading Paper in the Internal Printer section in the Accessories chapter for instructions on installing the printer Setting the utility If you wish to use a printer you can output to parameters for the printer e Internal printer e External RS232 prin
58. dditional parameter lines are added to the screen above the Cell Correction line If the Cell Correction line is no longer visible on the screen highlight More Parameters and press Enter a 4 Highlight Setup Correction and press Enter 5 Highlight Correction Mode and press Enter to set the mode to either Scan Cell Correction is run on a blank and one sample cell for the range of wavelengths you specify in Scanning mode Discrete nms The Cell Correction program is run on a blank and up to five sample cells for up to 31 user specified discrete wavelengths Press ft or to select item to change Run Set nms Came 6 If you selected Scan mode in the preceding step specify the Start Wavelength and the Stop Wavelength values 7 Press Run Corr to start the Cell Correction program If you selected Discrete nms mode first specify the wavelengths using the procedures which follow and then the Cell Correction program Evolution 60 User s Guide 41 42 Specifying wavelengths for Discrete nms mode Evolution 60 User s Guide The Cell Correction program will measure the other cells against the blank and will record store and date the measurements From these measurements the Cell Correction program establishes the required correction factors which then are automatically applied during all subsequent tests if Cell Correction is activated L Highlight Sample Positioner and press Enter to set this pa
59. diate measurement If you select more than one test as SmartStart tests the instrument when powered on will automatically display a menu that contains only those tests selected as SmartStart Note You will still be able to access the default main menu by pressing Test a Setting up a single 1 Press Utility to display the Utility screen test SmartStart tility 11 04 28Feb07 Software Revision 0 014b 2 Highlight Stored Tests Directory and press Enter Thermo Scientific Evolution 60 User s Guide 34 Thermo Scientific To unselect a test Utility Test Directory 14 56 21Feb0 7 ucleic Acid Tests DNA 260 280 Read onl DNA 260 230 Read only DNA with Scan 260 280 Read only DNA with Scan 260 230 Read only dsDNA Read only ssDNA RNA Read only Oligos Centered factor Read only Oligos calc factor Read only Direct UV 260 Read only Protein Tests Bradford Standard Read only Bradford Micro Read only Lowry Standard Read only ore tests Press fT or to select test Highlight the appropriate test and press Select Test to add the test to the SmartStart menu An arrow sign gt will indicate the test has been selected for the SmartStart menu You can now either press Esc to return to the Utility screen or you can power down the instrument Press Utility Highlight Stored Tests Directory and press Enter Highlight test to be removed from SmartStart menu and press Unsele
60. e absorbance and concentration The oligonucleotide calculator determines the following data for a base sequence that you enter Number of bases Percent GC content Molecular weight Molar Absorptivity or Extinction Coefficient Conversion factor to convert nucleotide absorbance at 260 nm concentration Tm for oligos of up to 40 bases for DNA DNA DNA RNA and RNA RNA hybrids Refer to the Parameters chapter for a description of the parameters and the Calculations for Oligo Calculator chapter for the default values To get started highlight Oligo Calculator and press Enter The Oligos screen appears Dligos 9 09 2Apr02 Base Sequence Number of bases GC Mol Wt 260 Conversion Factor Sis a aa Sequence Calc L 2 Press Base Sequence Select the appropriate character for the base you want to enter Thermo Scientific Thermo Scientific 3 Press Add Base to add the base to the sequence 4 When the base sequence is correct press Accept Sequence to accept it The instrument calculates and displays the results Dligos 9 19 2Apr02 ase Sequenc CATGTCGTTCTCAAACAGT Number of bases 20 GC 45 00 Mol Wt 6121 260 204730 Conversion Factor 2 base m Sequence Calc 5 To determine the theoretical Tm of the sequence press Tm Calc The Tm calculation screen appears Dligos 9 25 2Apr02Z Base Sequen bCATGTCETTCTCAAACAGT olarity of catio
61. e absorbance at each wavelength and displays it on the screen If you have set the measurement mode to Concentration Factor the instrument also displays the calculated concentration at each wavelength If a 6 Position Cell Holder is installed press Cell Position to reposition the cell holder and measure the rest of the unknowns manually Evolution 60 User s Guide 105 Performance Verification The Performance Verification program allows you to run tests to check the performance of your instrument Available tests include e Wavelength Accuracy Internal e Wavelength Accuracy Standards e Wavelength Repeatability e Resolution e Photometric Accuracy e Noise Measurement e Stray Light e Internal Printer Test e RS232C Test Running the appropriate performance verification tests regularly and maintaining a log of your results helps document the reliability of the instrument and indicates potential performance issues Note If you have a printer installed and turned on the instrument automatically prints out the test results for each performance verification test that you run You can also press Print to print another copy of the results a Thermo Scientific Evolution 60 User s Guide 106 Accessing the Performance Verification tests Troubleshooting Thermo Scientific checklist L Press Tests Highlight Performance Verification and press Enter Performance Verification 16 10 2Mar07 Wavelen
62. e calibration certificate included with your standards When running the Wavelength Accuracy test remember that You will need Wavelength Accuracy standards that are designed to measure the wavelength accuracy at the specified wavelengths An empty cell holder should be used as the blank The standards need to be measured in the order that they are listed on the test screen 1 5 standards can be used Highlight Wavelength Accuracy Standards and press Enter Evolution 60 User s Guide 109 Adding wavelengths L If you need to add a wavelength press Add nm and enter the wavelength value in the Entry field 2 When the value is correct press Add nm again to add the wavelength to the list The screen changes to prompt you to enter the tolerance for the wavelength you just entered Entry Enter Tolerance 1 0 20 0 Had trorcrancol 3 Enter the tolerance value in the Entry field 110 Evolution 60 User s Guide Thermo Scientific 4 When the value is correct press Enter to add the tolerance value to the list Deleting wavelengths p m If you want to delete a wavelength highlight the appropriate wavelength 2 When the appropriate wavelength is highlighted press Delete nm to delete the wavelength _ m Running the test Verify that the wavelengths and tolerances are set correctly 2 Press Start Test to run the test The results of the test appear on the screen indicating pass or fa
63. edit screen appears you can e Use all the data to recalculate the results e Select specific start and end times for the rate calculation and recalculate the results Kinetics Test Name KINETICS Abs 0 0 0 0 0 0 0 IDH min Abs Result 1 0 000 0 013 Press T or for more data eat creek Si data To use all the data to 1 With your table of kinetics data displayed on the calculate the edit screen press Use All Data reaction rate The instrument calculates the rate and displays it on the screen 82 Evolution 60 User s Guide Thermo Scientific To select specific start 1l With your table of kinetics data displayed on the and end times for the edit screen highlight the appropriate data point rate calculation in the table 2 Press Start Time or End Time The instrument displays the recalculated rate and result Thermo Scientific Evolution 60 User s Guide 83 Note Note Recalling a test Thermo Scientific Scanning When you use the Scanning program you can perform these tasks e Recall a test or set up new test parameters e Run the Cell Correction program e Collect a baseline scan e Scan an unknown e View scan data e Change the scale of the data plot e Determine 3 point net measurements e Calculate the area under a curve e Label peaks and valleys The Scanning program allows you to measure only one unknown at a time Auto 6 Auto 3 and Manual 6 are not available for sc
64. ees 106 Accessing the Performance Verification tests 107 Troubleshooting checklist ccccceeeeeeee essen eeeeeneeaas 107 Wavelength Accuracy internal ccceeeseee seen eens 108 Wavelength Accuracy Standards ccccceeeeeeee ees 109 Adding wavelengths sssssssssssrssssrenrssserenrsrserens 110 Deleting wavelengths ssssssssssrsrrsssrrrrrrssrrrrrresns 111 R nning the test nocni cirsio eds 111 Thermo Scientific Thermo Scientific Wavelength Repeatability cccccceeeeeeeeeeeeeeneeanes 111 RESOLUTION tecrtecnveee teres oevadet e E seven reed TEN 113 Photometric ACCUIACY ccceeesceseenseeeensseeennsseenees 114 Selecting the mode ovosscosvasvecsvecensavseerapureseanseceunde 115 Adding StandardS an eccsi once eevetiwn eta eines 115 Deleting standardS ssssrssssssrrrsssssrrrseensrrrrseens 116 Running the TES ssssssssssssssrrrrrrssrrrrrrenrrrrrrsssns 116 Noise Measurement scccissnsctiuneamn ra 117 Stray Lighter paneer oane NEE tenia 118 Running the test sssssssssssssssrrrrrssrrrrrresrnrrrrsssns 119 Internal Printer teSt ssssssrenrrernrrrrrrrnrrrnnrrrsnns 119 R5232 test orae a o E E A ea a 121 Bio Tests a a eee ees 123 SmartStart feature cccccccceeseeeseeeeeeseeeeesseeeeaaees 124 Setting up a single test SmartStart 000 124 Setting up a multiple test SmartStart 125 Nucleic acid measurement ssssssssrrrrs
65. entration Evolution 60 User s Guide 145 Cell growth Note Setting up test parameters Using the Correction Factor 146 Evolution 60 User s Guide The cell growth measurement uses absorbance at 600 nm to indicate the progress of cell growth in a sample The instrument does not perform any calculations or graphing for the data To get started highlight Cell Growth and press Enter The Cell Growth setup screen appears ell Growth 8 49 puu ID O OFF 1 ow High Limits 999979999 tatistics Off Press T or to select item to change ave ored un Test Tests Test If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Highlight the name of the parameter you want to set If you are using different spectrophotometers in your laboratory particularly if they are from a different manufacturer you may notice that readings for cell growth vary widely between these instruments This occurs because some instruments use different types of optical systems and detection schemes to collect data If you are using different spectrophotometers in your lab and would like to compare data from a different instrument use the correction factor option Thermo Scientific Measuring the sample Measuring unknowns automatically using Manual 6 or Single Cell Positioner Measuring standards manually using Manual 6 or Single Cell Positioner Thermo
66. entration the result appears on the screen along with the type of measurement the date and time the wavelength and the cell position used for the measurement To toggle between Absorbance Transmittance and Concentration press Change Mode until you see the desired mode is displayed You can toggle from one mode to another whenever you see Change Mode Absorbance 16 03 3iJan02 BLANK O 4 76A 660 Onm Ere as ee When Basic ATC is set to Absorbance or Transmittance you can perform these tasks e Set the wavelength e Measure a blank e Measure unknowns Evolution 60 User s Guide 46 Setting the 1 Press Set nm or any number key to set the wavelength wavelength Absorbance 17 25 io O 219A Enn Enter Wavelength 190 1100 2 Enter the wavelength where you want the measurements taken then press Set nm again Place the blank in the cell holder pm Measuring a blank If your instrument has a 6 Position Cell Holder installed be sure to place the blank in the B position 2 If you want to enter an absorbance or transmittance value for the blank press a number key and enter the desired value in the Entry field 3 Press Measure Blank to measure the blank When the instrument is finished measuring the blank the message disappears Thermo Scientific Evolution 60 User s Guide 47 Measuring unknowns 48 Basic Concentration measurements Evolution 60 User s Guide If you have
67. ermo Scientific Measuring unknowns j Absorbance Ratio 13 14 14Feb02 Measuring unknowns weet fang An BLANK automatically using Auto 6 or Auto 3 ID Abs Abs Abs 660 0 640 0 Ref WL Result Insert samples 1 5 into positions 1 5 Press ENTER to measure e ee ee ee Measuring unknowns manually L using Manual 6 or Single Cell Holder Thermo Scientific Place the blank and unknowns in the correct cell positions Press Enter to start the measurement The instrument automatically measures the blank first then measures the unknowns and then displays the sample measurements on the screen With the Absorbance Ratio screen displayed press Run Test Absorbance Ratio 13 22 14Feb02 Test Name Cell 1 Abs Abs Abs ID 660 0 640 0 Ref WL Result a a Blank Place the blank and unknown in the correct cell position If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder Evolution 60 User s Guide 71 3 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually 72 Evolu
68. ess Delete nm The Multiwavelength acquisition can be accessed from either the Set nms screen shown above or from the Evolution 60 User s Guide 103 Taking measurements manually using Manual 6 or Single Cell Holder 104 Evolution 60 User s Guide Multiwavelength set up screen 1 Press Run Test The Multiwavelength measurement screen appears Multiwavelength 4 46pm 18Feb02 Test Name BLANK ID 1 Result nm Abs Insert sample 1 into position 1 Press ENTER to measure Place the blank and the unknowns in the correct cell positions Press Enter to start the measurement The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen With the Multiwavelength screen displayed and the parameters set press Run Test Place the blank and unknowns in the cell holder If a 6 Position Cell Holder is installed be sure to place the blank in the B position You can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank Thermo Scientific Thermo Scientific When it completes the measurement it returns to its previous cell position With the list of wavelengths and factors displayed press Measure Samples to measure the unknowns The instrument measures th
69. ess ENTER to measure E Ss as ae 1 Place the blank and unknowns in the correct cell positions Evolution 60 User s Guide 133 Measuring unknowns manually using Manual 6 or Single Cell Holder 134 Evolution 60 User s Guide 2 Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen dsDNA 9 35 30Apr0Z Test Name dsDNA Cell f 1 Abs dsDNA IDH 260nm pg mL 1 0 426 21 30 Page 1 Sample 1 inane eT Bs Sample dsDNA 9 42 30Apr02Z Test Name dsDNA Abs dsDNA IDH 260nm pg mL a 1 Press Run Test 2 Place the blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder 3 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Thermo Scientific 4 Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually Oligos The Oligos calculated factor measurement calculates calculated factor molecular weight extinction coefficient and a concentration factor for a short lt 40 mer oligon
70. eypad not present on PC only models Sample Compartment Optional printer housing Thermo Scientific Evolution 60 User s Guide 2 Thermo Scientific Connectors The following illustrations show the locations of the connectors on the back of the spectrophotometer Key OAWOOO RS232C port Parallel printer port A C power connector On Off switch Fuse compartment A Warning Avoid shock hazard Always turn off the instrument and unplug it from the wall outlet or power strip before you unplug the power cord from the instrument connector a Evolution 60 User s Guide 3 4 About the keypad Evolution 60 User s Guide EVOLUTION 60 Absorbance Difference ength 2 Ref Wavelength Correction Factor 1 Factor 2 Units Sample Positioner Hunter of Samples 12 at 3Jan ag 2 oe SS ERFT 33 wo 885 Pres it lect iten t ee Save Stored TS Test Tests Test Key or Button 0000 Function Called Function keys Allows you to perform a specific function displayed on the screen above the appropriate key Functions change from one screen to another Depending on the application some function keys will not be active Can clear value being entered Does not change any values you have already accepted May return to the previous screen Deletes last character entered Does not change any values you have already accepted Does not return to the prev
71. gression curves first degree polynomials Linear regression through zero model pE poe SXY 3 N 1 Absorbance AbsA AbsA ADS ratio SS Absd Abs ADS Absorbance Result Difference ea AbsA factor 1 Absd factor 2 or Abs AbsAjpy factor 1 Abs Abs s factor 2 3 Point Net Baseline corrected absorbance A A A A 4 ls A 7 A 3 1 ABSORBANCE WAVELENGTH 3 Point Net Absorbance sample curve Thermo Scientific Evolution 60 User s Guide 169 Calculation Calculation s Graphs 3 Point Net ASTM E169 04 Baseline corrected absorbance A A A A La A A A 170 Evolution 60 User s Guide Thermo Scientific Calculations for Bio Tests Software Test Name Calculation s Default Displayed Parameters Results DNA Protein o diluent vol sample volume Ai 260 nm g mL 260 280 Dilution Factor D1 A gt 280 nm sample volume optional DNA concentration A A MN A A D i EATA f 50 Protein concentration A A 4 A A D f 1 dil vol 0 A Aye smp vol 1 Ratio A ap Aef DNA n diluent vol sample volume A 260 nm g mL 260 230 Dilution Factor D1 A gt 230 nm sample volume Ae 320 nm DNA concentration A Ang Mi A SA D fi re f2 3 48 Protein concentration
72. gth Accuracy Internal avelength Accuracy Standards avelength Repeatability Resolution Photometric Accuracy oise Measurement Stray Light Internal Printer Test IRS232C Test If a Performance Verification test fails follow the instructions below to help you diagnose common problems that may cause a test to fail If a test continues to fail after you have tried all the recommendations in the list follow the troubleshooting list for the test being run included with the description of each test Make sure you are following the instructions for the test properly Make sure filters and standards are clean Make sure the sample compartment door is closed while the test is running Make sure the sample compartment is clear of any obstructions Make sure the cell holder assembly is installed properly If the 6 Position Cell Holder is installed run the test once with the sample compartment door open to verify that the 6 Position Cell Holder is moving smoothly Evolution 60 User s Guide 107 A Warning A Warning Wavelength Accuracy internal 108 Evolution 60 User s Guide e Turn the instrument power OFF and then ON verify that no problems are indicated by the power on diagnostics e Make sure the lamp is ON e Make sure the lamp compartment is clear of any obstructions Do not open the lamp compartment unless the instrument power is OFF a Do not turn the instrument power ON unless the
73. gth you just entered Enter the T value in the Entry field When the T value entry is complete press Enter to add the T value to the list The screen changes to prompt you to enter the tolerance value for the wavelength you just entered Enter the tolerance value in the Entry field When the tolerance value entry is complete press Enter to add the tolerance value to the list The screen returns to the test screen displaying the values you just entered for that standard Press Start Test to begin the measurement or Press Esc to save test Evolution 60 User s Guide 115 Deleting standards L If you need to delete a standard highlight the appropriate standard 2 Press Delete Std to delete the standard from the list Running the test With the Photometric Accuracy screen displayed make sure that the wavelengths absorbance or T values and tolerances are set correctly 1 Press Start Test to run the test The results of the test appear on the screen indicating pass or fail for each wavelength Standards 1 2 in positions 1 2 Start Test ESC to save test ange Add to Abs Std If the test fails follow these guidelines e Repeat the test twice to verify that the test is failing consistently e Make sure you follow the guidelines provided with the standard reference materials e Contact technical support for more troubleshooting advice 116 Evolution 60 User s Guide Thermo Scientific Noise Me
74. he Math screen appears you can e Smooth data e Determine 3 point net e Calculate the area under a curve e Label peaks and valleys Scanning 9 15 15Feb02 Test Name FILTER BLANK 2 220 0 070 50 0 Wavelength 500 0 ID 1 Baseline collected 15Feb02 Seman hor fren UNE COffl Net Valleus Smoothing data If your scan shows sampling noise you can smooth the data with the smoothing function 1 With your scan data displayed on the edit graph screen press Smoothing On The instrument performs a data smoothing function on the data and displays the smoothed graph 92 Evolution 60 User s Guide Thermo Scientific Determining 3 point net measurements Thermo Scientific L 2 With your scan data displayed on the edit graph screen press Math Press 3 Pt Net to enter the 3 point net measurement screen A screen showing the cursor options and three cursor lines designated for the left center and right wavelengths appears Scanning 13 45 20Jun02 Test Name Filter 407 415 425 0 045 0 384 0 041 0 400 A 020 Wavelength 435 B Pt Net Value 0 341 Factor 1 000 nter Nex gt Factor Cursor Cursor Cursor Use Cursor and Cursor lt to position the left cursor line to the desired wavelength value The instrument calculates the 3 point net absorbance for the selected wavelengths Continue selecting the other wavelengths by pressing Next Cursor to activate the center and
75. he blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually Evolution 60 User s Guide 143 Warburg Christian The Warburg Christian analysis uses an absorbance difference measurement at 280 and 260 nm to determine the protein concentration of an unknown Refer to the Parameters chapter for a description of the parameters and Calculations for Bio Tests Software chapter for the default values To get started highlight Protein Tests and press Enter Then highlight Warburg Christian and press Enter The Warburg Christian parameter screen appears Marburg Chr istian 13 05 i Christian Default avelength 1 280nm avelength 2 260nm actor 280 1 55 actor 260 0 76 Dilution Multiplier calculated 1 000 nits C Sample Positioner Auto 6 umber of Samples 1 More parameters Press f or to select item to change a a Test Tests Test Note If Cell Correction is ON you must run the Setup Correction prog
76. he wavelengths you wish to measure press Save Test to save the test or Run Test to measure a blank or unknowns If you have not selected the wavelengths you can add wavelengths and factors as shown below Note If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Note If Auto Save Data is ON you must enter a Data File Name before you can access Run Test or Measure Samples a Adding wavelengths and factors Note You can enter factors only when the measurement mode is set to Concentration Factor a 1 Press Set nms 102 Evolution 60 User s Guide Thermo Scientific Deleting wavelengths and factors Note Taking measurements Taking measurements automatically Auto 6 or Auto 3 Thermo Scientific Multiwavelength Test Name WLH nm 1 610 0 2 620 0 3 680 0 Page 1 Press f or to view data Edit ki 18Feb02 aa ee Test 2 Highlight the position where you want to enter the first wavelength and factor pair 3 Press Add nm 4 Enter the values for the wavelength and factor and press Enter 5 When the values are correct press Add nm 6 Continue until you have entered all the wavelengths and factors m Highlight Set nms If no wavelength values have been entered the wavelength and factor columns will be empty a 2 Highlight the first wavelength and factor pair you want to delete 3 Pr
77. ign gt will indicate the test has been selected 4 Press Load Test The parameter screen of the test you selected will be displayed At this point you can power down the instrument and then power it back up When it starts up again the parameter screen for the selected test will be displayed a Thermo Scientific Setting up a multiple test SmartStart Note Nucleic acid measurement Thermo Scientific 1 Press Stored Tests 2 Scroll through the list until the first appropriate test is highlighted 3 Press Select Tests to add the selected test to the SmartStart menu 4 Continue scrolling through the list and adding tests until all the appropriate selections are made 5 Press Esc until you return to the Bio Tests screen At this point you can power down the instrument and then power it back up When it starts up again the list of tests you ve selected will be displayed a You can use these tests to determine the concentration and purity of nucleic acid in a given sample DNA Direct 260 Measures the DNA concentration by direct absorbance at 260 nm An optional background correction at 320 nm is available DNA 260 280 Measures absorbance at 260 and 280 nm determines DNA concentration and purity based on absorbance ratio and absorbance difference An optional protein concentration calculation and background correction at 320 nm is available DNA 260 280 with scan Records absorbance us
78. il for each wavelength If the test fails follow these guidelines e Repeat the test twice to verify that the test is failing consistently e Make sure the target value you entered for the calibrated wavelength is the same as the value printed on the calibration certificate for the standard e Make sure the tolerance value you entered for the calibrated wavelength is the same as the value given on the calibration certificate for the standard Wavelength This test measures the ability of the spectrophotometer Repeatability to return to an identical wavelength in a repeatable manner The test uses the internal xenon lamp e A xenon lamp has a strong fundamental line 529 nm This line is an essential property of xenon and serves as a fundamental standard When running the internal standard test remember that Thermo Scientific Evolution 60 User s Guide 111 112 Evolution 60 User s Guide The wavelength and tolerance values are preset and cannot be changed The cell holder should be empty Highlight Wavelength Repeatability Press Enter The Wavelength Repeatability screen appears Press Start Test to run the test The results of the test appear on the screen indicating pass or fail for each wavelength erformance Verification 16 33 2Mar07 avelength Repeatability inimum Maximum Peak to Peak Result 29 2 529 3 0 1 Pass ake sure the Blank position is empty Press Start Test If the test fails f
79. ime options that you can modify date time format and time L Highlight Set Date and press Enter 2 Press Set Day type the date and then press Enter Thermo Scientific Utility 3 40pm 16Mar07 Time Format Set Time Entry Enter Day 1 31 3 Press Set Month highlight the correct month and then press Enter Dec Press f or to select Press ENTER Press ft or 1 t Enter Entry Enter Year 0 99 5 When the date is correct press Esc to save the settings and return to the Utility screen Thermo Scientific Evolution 60 User s Guide 23 To select time format To set the time 24 Evolution 60 User s Guide You can set up the spectrophotometer to display the time in either am pm format or in 24 hour format To change the display format for the time highlight Time Format and press Enter until the format that you want to use AM PM or 24 hour appears L Highlight Set Time and press Enter 2 To set the hour press Set Hour type in the hour and press Enter 3 42pm 16Mar07 Utility Entry Enter Hours 1 12 3 To set the minutes press Set Minute type in the minute and press Enter Utility 3 43pm 16Mar07 Entry Enter Minutes 0 59 4 To select between AM and PM if in AM PM time format press Set AM PM until the appropriate setting appears Thermo Scientific Note Selecting standby setting
80. in your instrument You can install only three each of some accessory cell holders Make sure to place them in position B 2 and 4 a Follow the steps below to install the optional internal printer Turn off the instrument and disconnect the power cord from the outlet before installing the internal printer a Evolution 60 User s Guide 15 16 WM Key Captive screw on lamp door Removing door from hinge Hinge Connecting wires to the printer 1 Loosen the captive screw on the lamp door by rotating it counterclockwise about turn 2 Open the lamp door 3 Use a pen or screwdriver to lift the tabs holding the door to the hinge and 0 4 Slide the door off the hinge 5 Remove the printer already installed on the printer door from its packing 6 Unclip the connector wires on the door hinge 7 Lower the hinge so it is out of the way 8 Connect the wires and press into place with a small screwdriver Evolution 60 User s Guide Thermo Scientific Loading paper in the Thermo Scientific internal printer 10 11 12 There is only one way that the connectors will fit Each connector has a slight D shape Make sure the side of the connector with the shiny metal contacts faces away from the printer and towards the plastic door Use the clip on the hinge to secure the wires Install the printer door by sliding it bac
81. ing a scan between 225 and 325 nm determines DNA concentration and purity based on absorbance difference An optional protein concentration calculation background correction at 320 nm is available Evolution 60 User s Guide 125 126 Evolution 60 User s Guide DNA 260 230 Measures absorbance at 260 and 230 nm determines DNA concentration and purity based on absorbance ratio and absorbance difference An optional protein concentration calculation and background correction at 320 nm is available DNA 260 230 with scan Records absorbance using a scan between 225 and 325 nm determines DNA concentration and purity based on absorbance ratio and absorbance difference An optional protein concentration calculation and background correction at 320 nm is available dsDNA Measures absorbance at 260 nm calculates concentration based on absorbance and concentration factor ssDNA RNA Measures absorbance at 260 nm calculates concentration based on absorbance and concentration factor Oligonucleotides Oligos Measures absorbance at 260 nm calculates concentration based on absorbance and concentration factor or calculates concentration based on absorbance and concentration factor determined by the oligo calculator Several of these categories include multiple tests that are similar so the section may not include screen samples for each test For example the parameters are the same for the Direct UV measurement of ssDNA and RN
82. ious screen Thermo Scientific Key or Button Function e Accepts highlighted or selected values e Advances to next parameter or screen e Prints the displayed information e Displays the Test Types screen e Displays the Utility Screen OOO e Controls the location of the cursor e Highlights the value or option for selection e Enters numbers decimal point and minus sign for entry of values 000000 0000 O O Q e Cell position keys e Selects cell holder position to be measured e B blank and 1 5 sample positions i positions when used in Auto 3 mode Thermo Scientific Evolution 60 User s Guide 5 Cell Holders All models of the Evolution 60 include a 6 position and single cell holder 6 Position Cell Holder Single Cell Holder Note Whenever you press Run Test in any of the test modes to start a measurement the instrument will attempt to initialize the cell positioner If a single cell holder is installed the message Error Single Cell Holder found Use Single Cell Holder is displayed Press Accept Change to continue or install the cell changer and press Cancel Change a For a more detailed list of accessories available please see the parts list 6 Evolution 60 User s Guide Thermo Scientific Selecting and positioning cuvettes Test tubes vs cuvettes Thermo Scientific Other guidelines The compatible wavelength range for different types of cells depends upo
83. ivated Thermo Scientific Evolution 60 User s Guide 43 Auto 6 Auto 3 Single Cell Holder Thermo Scientific Note Sample Positioner Setting The spectrophotometer lets you use different cell holders and cell holder accessories to take measurements When you select your test parameters you will select the type of measurement you require and indicate how many samples you have You can choose from the following measurement options Take one blank measurement and up to five sample measurements without changing the samples in the cell holder The instrument automatically measures the blank in the blank position then automatically advances the cell holder to the appropriate position for the next measurement Take up to three measurements without changing the samples in the cell holder The instrument automatically measures the blank in the blank position then automatically advances the cell holder to the appropriate position for the next measurement position 2 or 4 Place the blank in the cell holder measure it place a sample in the cell holder then measure your sample This process is completely manual In fact the cell position buttons do not function when you select Single Cell Holder You can use the 6 Position Cell Holder as a single cell holder by selecting this option However measurement error can occur if in the process of removing and inserting the cell you move the position Therefore the Single Cel
84. k onto the hinge Close the lamp door Tighten the captive screw on the printer door to hold it securely in place Key O Paper roll holder Icon for paper direction Paper entry slot Finger tab Evolution 60 User s Guide 17 Note Make sure that the paper roll holders are in place as shown When installed correctly they will fit flush with the top of the instrument a 13 Cut the paper so the edge is even Note Arrows on the paper roll holders indicate the direction of the paper feed a 14 Feed the paper straight into the paper entry slot The printer grabs the end of the paper and pulls it in 15 In Basic ATC mode when the paper stops press Enter to continue advancing the paper until the paper comes out of the paper exit slot 16 Pull out on the finger tabs on the paper roll holders and secure the roll of paper onto the paper roll holder 18 Evolution 60 User s Guide Thermo Scientific Thermo Scientific External printers Note Note Note Your spectrophotometer is able to print to external desktop printers supporting HP PCL 5 0 format and later PCL format does not support HP Windows printers a To print to an HP PCL printer connect the parallel interface cable to the printer and to the parallel port on the back of the instrument Connectors section The instrument is compatible with specific printers and printer cables we offer Contact technical support or you
85. king Turret Please Wait while it attempts to initialize the cell changer at the Blank position If a single cell holder has been installed the message Error Single Cell Holder found Use Single Cell Holder is displayed Press Accept Change to continue or install the cell changer and press Cancel Change Evolution 60 User s Guide 11 12 Note Changing cell holders Evolution 60 User s Guide If the 6 Position Cell Holder has been installed it will initialize it to the B position After it has initialized the cell changer the instrument will display the data collection screen for the test If while in this screen you remove the cell changer and press Run Sample the instrument will display Fatal Error 8 Press Esc to return to main menu Pressing Esc returns you to the parameter menu screen of the test Press Run Test and the instrument will now display the message Calibrating and Checking Turret Please Wait while it attempts to initialize the cell positioner at the Blank position If the cell changer is still not in the sample compartment the instrument will display Error Single Cell Holder Found Use Single Cell Holder Either press Cancel Change and reinstall the cell changer or press Accept Change to continue with using the single cell holder To prevent the Fatal Error whenever removing the cell changer always return to either the main menu or the test parameter menu of the test
86. l Holder is recommended for accessories requiring a high degree of positioning repeatability such as the nanoCell accessory small aperture microcells the coupling module of fiber optics probes and flowcells used with a sipper system a Evolution 60 User s Guide 44 Thermo Scientific Manual 6 Note Take up to six measurements without changing the samples in the cell holder using the cell position buttons to advance the cell holder to the appropriate position for the next measurement You place the blank in the blank position and your samples in the other cell positions Regardless of where the cell holder is positioned when you press Measure Blank the cell holder automatically goes to the blank position and measures the blank However you can use the cell position buttons to select a different position for the measurement When you have the 6 Position Cell Holder installed the instrument always considers the material in the B position as a blank This means that even after measuring your blank the first time you can place unknowns only in positions 1 through 5 a Evolution 60 User s Guide 45 Thermo Scientific Basic ATC Mode Abs amp T Measurements The Basic ATC mode puts the instrument into an instant measurement mode The lamp flashes continuously so that you can walk up to the instrument insert your sample and measure it Depending on whether the mode is set to Absorbance A Transmittance T or Conc
87. lamp compartment is closed a This test locates the peaks of the internal xenon lamp and displays the expected and measured wavelengths for the peaks e A xenon lamp has strong fundamental lines at 229 529 and 883 nm These lines are an essential property of xenon and serve as a fundamental standard When running the internal standard test remember that e The wavelengths and tolerance values are preset and cannot be changed e The cell holder should be empty 1 Highlight Wavelength Accuracy Internal 2 Press Enter The Wavelength Accuracy Internal screen appears 3 Press Start Test to run the test The results of the test appear on the screen indicating pass or fail for each wavelength Thermo Scientific Thermo Scientific Wavelength Accuracy Standards erformance Verification 16 13 2Mar07 avelength Accuracy Interna a ah Tolerance Measured Result m n nm 47 5 1 248 0 Pass 29 0 1 529 3 Pass 24 2 1 824 4 Pass ake sure the Blank position is empty Press Start Test If the test fails follow these guidelines Repeat the test twice to verify that the test is failing consistently Contact technical support for more troubleshooting advice This test measures the absorbance or T of a wavelength accuracy standard at up to five wavelengths and compares the results with specified tolerances The wavelengths and tolerances are preset but you should change them to the values on th
88. lculated factor Entering a factor Highlight Factor If you need to change the factor enter the correct factor If you need to change the units highlight Units and select the correct units 58 Evolution 60 User s Guide Thermo Scientific Measuring unknowns Measuring unknowns automatically using Auto 6 or Auto 8 Measuring unknowns manually Thermo Scientific using Manual 6 or Single Cell Holder pm m Press Run Test The Advanced A T C measurement screen appears prompting you to place your samples in the cell holder Place the blank and unknowns in the correct cell positions and press Enter The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen Press Measure Sample to measure additional unknowns Press Run Test Place the blank and unknown in the correct cell position If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to reposition the cell holder and measure the rest of the unknowns manually Evolution 60 User s Guide
89. le Cell Holder is installed press Measure Blank to measure the blank then insert your unknown and press Measure Sample When the instrument completes the measurement the kinetics data and rate appear on the screen Kinetics 3 05pm 27Jan01 Test Name ENZYME Gell 1 0 740 A 0 583 Time 3 00 IDH A min Result 1 0 050 22 69 di Measure tabular Graph Sample _ If you want to switch between tabular and graphical displays press Graph Tabular Thermo Scientific Recalling and recalculating graphical kinetics Thermo Scientific results When you have a graphical display you can press Edit Graph and then press Cursor to move the cursor line from one position to another on the plot As the cursor moves the rate and result values indicate the values for the point on the plot where the cursor is located Within the Kinetics program you can view and manipulate your results either in graphical form or in tabular form When your results are displayed on the screen you can modify the range start and stop time and the instrument automatically recalculates the reaction rate When you are working with graphical kinetics results you need to press Edit Graph before you can rescale and recalculate Kinetics 3 29pm 18Feb02 Test Name ENZYME Cell 1 41 850 A 0 840 Time 3 00 IDH Abs nmin Result 1 0 251 1756 Cursor Scale eniai Data You can modify the scale of your kinetics data plot in t
90. me DNAscan 260 280 Cell 1 0 560 0 025 225 Wave length 325 ID DNA Ratio DNA Conc Protein Conc 1 1 803 21 60 52 12 Baseline collected 28Mar02 gina rbe a Blank Sample These test measurements are all set up and run using the same types of test parameters Refer to the Parameters chapter for a description of the parameters and the Calculations for Bio Tests Software chapter for the default values To get started highlight Nucleic Acid Tests and press Enter A list of nucleic acid test appears Highlight the desired test and press Enter The dsDNA ssDNA RNA or Oligos entered factor parameter screen appears Thermo Scientific Note Note Setting up test parameters Measuring unknowns Measuring unknowns automatically using Auto 6 or Auto 3 Thermo Scientific The following screens show the parameters for the dsDNA test a If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a 11 11 28Mar02 Cell 1 Sample Positioner umber of Samples ore parameters Press T or to select item to change L Highlight the name of the parameter you want to set 2 When the parameters are set press Save Test to save the test under a new name or Run Test to measure the blank and unknowns AsDNA 9 04 30Apr02Z Test Name dsDNA BLANK Abs dsDNA IDH Z60nm pg mL Insert sample 1 into position 1 Pr
91. measure concentration 2 3 4 5 Thermo Scientific Press Measure Standard to measure the standard When the instrument is finished measuring the absorbance of the standard it displays the absorbance and calculated factor If necessary press Change Mode to switch to the Conc With Factor mode Press Units Factor to set the factor and select the units Conc with Factor 15 29 6Mar01 BLANK Factor 45 Units C Press T or to select item to change Gas ro a Factor Units Press Enter Factor Type the desired factor value Press Enter Factor to accept the factor and return to the screen displaying the factor and units Evolution 60 User s Guide 51 Conc Factor 11 18 4Feb02 Factor 89 4 BLANK OP Ong mL 660 Onm ister pascal Saag Blank Factor Mode 6 Press Select Units 7 Highlight the appropriate unit in the list and press Enter to select the units for the concentration 8 Press Esc to return to the Conc With Factor screen Measuring unknowns if the 6 Position Cell Holder is installed place the unknown you want to measure in one of the cell positions and press the corresponding cell position button to move the cell holder to the measuring position The measurement appears on the display If the Single Cell Holder is installed remove the blank and place the unknown in the cell holder The measurement appears on the display 52 Evolution 60 User s Guide Thermo Scien
92. n 0 050 Formanide 0 Mismatch 0 Tm Cup to 20 bases only 58 0 Tm DNA DNA 53 1 Tm CDNA RNA 56 2 Tm CRNA RNA 62 7 oe a ae a Sequence Enter the formamide and mismatch if known that will be used to calculate the Tm The calculated Tm values are shown on the screen Evolution 60 User s Guide 149 Utility Calculator Function 1 From the Utility menu highlight Calculator and press Enter tility 16 10 6Mar07 Software R Language 4 of lamp Screen Con Printout C Stored Tes alculator 2 Use the numeric keypad to enter the desired value 3 Press the desired function x or 4 Enter the second desired value and press Enter to calculate the total Note You can only add subtract multiply or divide two lines of numbers at a time a tility 9 23am 14Mar07 Screen Con Printout C Stored Tes alculator Thermo Scientific Evolution 60 User s Guide 150 Thermo Scientific A Warning Routine care Maintenance The spectrophotometer is durable and reliable so routine maintenance is minimal This section includes complete instructions for e Routine care cleaning and maintenance of the instrument and cells e Changing the fuse and voltage setting Operating the instrument with the cover off exposes the operator to potentially dangerous voltages and ultraviolet UV radiation Therefore we recommend that only authorized servi
93. n of a standard sssssssssess 66 T oadd a standardar enean a ea 66 To delete a standard sssssssssnsusnrusnrsnrnnrrrrrsnrenrnsnns 67 TO Clear meas reM ntS secre a 67 To reset standardS s sssssssnrssrssrsnrrurnnenrrnrsnrenrrrns 67 To select a different curve fit for a standard curve 67 Absorbance Ratio cccscsssssssssssssessesseseesessesstssessesssaesatseesessseacsacsaess 69 Recalling a testean oireann nan a aA 69 Setting up test parameters ccc cece cece eceeeeeeeeeeeeeeees 70 Measuring UNKNOWNS ci ecntus teresa ied icgecetecs Gente w ies 71 Absorbance Difference c cccccssssssssesssssssessssssssssseseesessesaeeaeseess 73 Recalling a testaria dia ee es 73 Setting up test paraMeters cece eee e ccc eeeeeeeeeeeeees 74 Measuring UNKNOWNS ceeceee eee tenet ee eeee ee nnnee enna eens 74 Evolution 60 User s Guide iii iv Evolution 60 User s Guide Lg a 2h gt ee enti nae OR NER CRORE NE E RDO STON OR SPREE A CR SRT OM RE RTO 76 Recalling a Test 2 cc056s a dvtwrini eei a a 76 Setting Up test parameters ccc cccceeceee eee eee e ee eeeeeees 77 Measuring UNKNOWNS sssssssssssssssssrrrrsssrrrrrssssrrrresesnns 78 Recalling and recalculating graphical kinetics results 79 Auto Scale fUNCtON ieni rne a Ea E 80 Manual Scale function c cece ee ecee eee ee eseeeeeneneaes 80 CURSOR TUREUOMN ese ranea a a E 81 Rescaling and recalculating tabular kinetics results
94. n the material used Cell Type Wavelength Glass Optical Glass 360 to gt 1100 nm Special Optical Glass 320 to gt 1100 nm Pyrex 320 to gt 1100 nm Borosilicate Glass 330 to gt 1100 nm Quartz also called silica Herasil Quartz 230 to gt 1100 nm Infrasil IR Quartz 220 to gt 1100 nm Suprasil 200 to gt 1100 nm ES Quartz 190 to gt 1100 nm Spectrosil Far UV 170 to gt 1100 nm Disposable Polystyrene gt 340 nm Methacrylate gt 300 nm Acrylic gt 280 nm UV transparent Plastic gt 220 nm Refer to manufacturer s specifications and ensure that you work within the recommended range Square cuvettes yield very precise results However matched test tubes when properly handled can show as little as 1 2 deviation between readings The pathlength of test tubes is not as well defined as in square cuvettes However constructing a standard curve eliminates the need for great accuracy in knowing the pathlength provided that the same pathlength cell is used for all blanks standards and samples Be sure to position cuvettes and test tubes so that the clear sides face the light beam This means that one clear side should face the front of the instrument and the other clear side should face the back of the instrument Evolution 60 User s Guide 7 Note Z dimensions Cell holder configurations Evolution 60 User s Guide Test tubes should always be placed in the instrument in exactly the same orientation in
95. ncentration based on a factor An optional correction at 320 nm is available Bradford Measures absorbance at 595 nm determines concentration for either standard or micro sample concentrations Lowry Measures absorbance at 550 nm for standard and 770 nm for micro determines concentration for either standard or micro sample concentrations Bicinchoninic Acid BCA Measures absorbance at 562 nm determines concentration for either standard or micro sample concentrations Evolution 60 User s Guide 137 138 Evolution 60 User s Guide Note Note Biuret Measures absorbance at 540 nm Several of these categories include multiple tests that are similar so this section includes screen samples for the standard Bradford test only For a complete list of all parameters for each test refer to the Parameters chapter for a list of calculations used for the tests refer to the Calculations for Bio Tests Software chapter for default values To get started with the Bio Tests screen displayed highlight Protein Tests and press Enter A list of protein tests appears Highlight the desired test and press Enter The Bradford Standard parameter screen appears The following screens show the parameters for the Bradford Standard test All the other protein standard curve methods work in a similar fashion a Bradford Standard 16 40 28Mar02 Cell 1 Linear Regr 2nd Order umber of Standards nits pg mL Sample Positi
96. not active when the cell holder is set to 1 Cell or Manual 6 nor when the Single Cell Holder is installed a Before running the cell correction program e Clean the inside and outside of all the cells to be matched e Fill the cells with distilled water or other blank solution and place them in the sample compartment see Selecting and Positioning Cuvettes Be sure to place the blank cuvette in Cell B of the sample compartment Thermo Scientific Evolution 60 User s Guide 39 Running the 1 Load the test type or stored test Cell Correction program Advanced A T C Ref Wavelength Correction Ref Wavelength Delay Time min sec 0 00 Sample Positioner Auto 6 umber of Samples 1 More parameters Press fT or to select item to change ave ored Run Tests 2 If Cell Correction is not visible on the parameter list then highlight More parameters and press Enter 3 Highlight Cell Correction and press Enter Advanced A T C ell Correction Correction ID CO OFF 1 ow High Limits 999979999 tatistics Off More parameters Cell Correction Not Calculated Press T or to select item to change ored Tests The Cell Correction function is now activated as indicated by the word On across from Cell Correction on the test setup screen 40 Evolution 60 User s Guide Thermo Scientific Thermo Scientific Note When Cell Correction is activated a
97. ns to re position the cell holder and measure the rest of the unknowns manually Evolution 60 User s Guide 75 Note Note Recalling a test Thermo Scientific Kinetics When you use the Kinetics program you can perform these tasks e Recall a test or set up new test parameters e Run the Cell Correction program e Measure a blank e Measure an unknown e Recalculate reaction rates e Modify the scale of the plot When you use the Kinetics program you can choose to work with either graphical data or with tabular data You can perform the same functions regardless of the type of display you select However the location of the function keys varies depending on the type of display you choose The Kinetics program allows you to measure only one unknown at a time a The Kinetics program allows you to collect up to 400 data points per run When you set up your test parameters be sure to select the interval time and total run time accordingly a To get started press the Test button When the Test Types screen appears highlight Kinetics and press Enter L With the Kinetics screen displayed press Stored Tests 2 Highlight the name of the test you want to recall and press Enter Evolution 60 User s Guide 76 Setting up test parameters Note Note Thermo Scientific inetics 13 45 14Feb02 Cell 1 easurement Mode Absorbance ave length 546 Onm ef Wavelength Correction Off elay Time min sec 0
98. nstrument highlights the selected data points You can press Graph to display the plot using the highlighted data points Thermo Scientific Thermo Scientific Note Note 3 Point Net When you use the 3 Point Net program you can perform these tasks e Recall a test or set up new test parameters e Run the Cell Correction program e Measure a blank e Measure unknowns To get started press Test When the Test Types screen appears highlight 3 Point Net and press Enter B Point Net 10 11 15Feb02 BLANK BILIRUBIN Sample Positioner umber of Samples ore parameters Press ft or to select item to change If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a If Auto Save Data is ON you must enter a Data File Name before you can access Run Test or Measure Samples a Evolution 60 User s Guide 97 Recalling atest 1 Press Stored Tests 2 Highlight the name of the test you want to recall and press Enter The parameters for the selected test appear on the screen From this screen you can e Set up test parameters e Run the Cell Correction program e Save atest e View the list of stored tests e Measure a blank e Measure unknowns Setting up test 1 Highlight the name of the parameter you want to parameters set 2 When the parameters are set press Save Test to save the test or Run Test to measure a blank or
99. nt mode you select See the Parameters chapter for a complete list When the parameters are set press Save Test to save the test or Run Standard to measure a standard if in Conc Std or press Run Test if in Conc Factor With the Measurement Mode set to Conc Std press Run Standard Advanced A T C 10 17 20Feb02 Test Name STANDARD TEST BLANK Conc Abs Factor ppb 650 0nm calculated Entry Enter a number 0 001 to 9999 Enter the concentration value of the standard and press Enter Press Measure Blank Place the blank and standard in the correct cell positions Evolution 60 User s Guide 57 5 Press Enter to measure the blank and the standard The instrument automatically measures the blank first then measures the unknowns and displays the absorbance and calculated factor Measuring a standard manually L Press Run Standard using Manual 6 or Single Cell Holder 2 Enter the concentration value of the standard and press Enter 3 Place the blank and standard in the correct cell positions 4 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 5 Press Measure Standard to measure the standard When the instrument is finished measuring the absorbance of the standard it displays the absorbance and ca
100. ollow these guidelines Repeat the test twice to verify that the test is failing consistently Contact technical support for more troubleshooting advice Thermo Scientific Thermo Scientific Resolution This test measures the ability of the spectrophotometer to resolve adjacent features in a spectrum The test is performed using a 0 02 v v solution of toluene in hexane and requires a hexane blank When running the internal standard test remember that the wavelengths and tolerance values are preset and cannot be changed 1 2 Highlight Resolution Press Enter Ensure that Hexane is in the blank position and Toluene in Hexane is placed in Cell 1 performance Verification 16 37 2Mar07 esolution Maximum Minimum Ratio Result 0 100 0 037 2 672 Pass Place Hexane in the Blank position Place Toluene in Hexane in Cell 1 Press Start Test Press Start Test to run the test The results of the test appear on the screen indicating pass or fail for each wavelength If the test fails follow these guidelines Repeat the test twice to verify that the test is failing consistently Contact technical support for more troubleshooting advice Evolution 60 User s Guide 113 Photometric Accuracy 114 Evolution 60 User s Guide Note This test measures the absorbance or T of a set of standards and compares the results with specified tolerances The wavelength absorbencies and tolerances are preset
101. oncentration a _ slope The slope is calculated as a SXY SXX This model requires Slope is not equal to zero or infinity At least one standard data point with concentration gt 0 The absorbance of the 0 concentration blank OA Segmented The segmented model requires model Data for at least two standard data points with different concentrations and absorbances Slopes of all segments must be ascending positive or descending negative Validity of A c gt Alc for all c gt c NY standard IW a curves or a ORE e e MG Alc lt Alc forall c gt c 4 I where A absorbance concen az EN Ci C concentration N TRATION Ri S J gt Cmn C max CONCENTRATION DOMAIN CONCENTRATION Valid non linear standard curve If this is not the case then there will be more than one solution within the specified domain and the message Curve cannot be used to determine unknown concentrations it may produce ambiguous results will appear when the curve is viewed ABSORBANCE a CONCENTRATION Invalid non linear standard curve 168 Evolution 60 User s Guide Thermo Scientific Calculation Calculation s Graphs Statistics y2 Linear ae DAY J regression N n 1 general case where n Degree of polynomial ISSXY SOX SQY Note The calculation for the correlation coefficient only applies to first order linear re
102. oner Auto 6 umber of Samples ore parameters Press T or to select item to change kee e e Standards Test Tests If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Thermo Scientific Setting up test parameters for a standard curve Measuring the standards for a standard curve Measuring standards automatically using Auto 6 or Auto 3 Thermo Scientific 1 Highlight the name of the parameter you want to 2 L set Set the parameters for measuring the standards Refer to the list of parameters in the Parameters chapter for a description of the parameters and Calculations for Bio Tests Software for the default values When the parameters are set press Save Test to save the test or Run Standards to measure the blank and standards If you need to edit concentration values select the standard you want to edit and press Edit Standards From the Edit Standards window you can edit add or delete one or all standards Bradford Standard 10 38 1 pr02 Test Name Bradford Std BLANK Std Conc Abs Page 1 of 2 Standards 1 5 Press f or to view data pT RES Toate tema Test Standards Standards Place the blank and standards in the correct cell positions When all the standards are correct press Measure Standards to set up and run the standards Evolution 60 User s Guide 139 Measuring standards m
103. ou use the Absorbance Ratio program you can perform these tasks e Recall a test or set up new test parameters e Run the Cell Correction program e Measure a blank e Measure unknowns To get started press Test When the Test Types screen appears highlight Absorbance Ratio and press Enter Recalling a test 10 32 20Feb02 BLANK avelength 1 avelength 2 Ref Wavelength Correction off Sample Positioner Auto 6 umber of Samples 5 ore parameters Press T or to select item to change ana A Test Tests Test 1 With the Absorbance Ratio screen displayed press Stored Tests 2 Highlight the name of the test you want to recall and press Enter Thermo Scientific Evolution 60 User s Guide 69 70 Setting up test parameters Evolution 60 User s Guide Note Note From this screen you can e Set up test parameters e Run the Cell Correction program e Save atest e View the list of stored tests e Measure a blank e Measure unknowns L With the Absorbance Ratio screen displayed highlight the name of the parameter you want to set 2 When the parameters are set press Save Test to save the test or Run Test to measure a blank or unknowns If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a If Auto Save Data is ON you must enter a Data File Name before you can access Run Test or Measure Samples a Th
104. r sales representative for more information a If the printer cable is not purchased from us it is the responsibility of the customer to determine if the printer cable is compatible with the instrument a Evolution 60 User s Guide 19 Entering parameter Thermo Scientific values Numeric entry Setting up the Instrument This chapter provides important information about using your instrument to analyze samples For models with Local Control you will use the keypad to enter or select values This section describes the various modes of entering or selecting parameter values With the parameter e g Wavelength highlighted start typing the numeric value An Entry window with the value range appears Type the complete entry and press Enter Advanced A T C 17 02 28Feb0 BLANK Test Name leasurement Mode h Sample Positioner umber of Samples Entry g Enter Wavelength 190 1100 Alternatively you can press Enter to display the Entry window with the value range and then type the complete entry and press Enter Evolution 60 User s Guide 20 Alphanumeric entry Menu selection On Off toggle Setting utility Thermo Scientific parameters With the parameter e g Units or Sample Positioner highlighted press Enter The selection list will appear Highlight the appropriate item and press Enter Sample Positio umber of Samp Unit ore parameter Press fT or to select Press ENTER
105. r password to allow user to lock or unlock the file Utility Low High Limits Enter lowest and highest acceptable results outside of which the result is flagged as Low or High Adv A T C Std Curve Abs Ratio Abs Diff Kinetics 3 Pt Net and some Bio Tests Math Accesses manipulation functions of graph Scanning Measure Blank as function key Initiates measurement of blank Measure Blank as test parameter Selects frequency of zeroing the instrument Once or Every Reading Kinetics Measurement Mode Selects the type of photometric data reported for a measurement Abs T Conc in A T C Kinetics Scanning Multiwavelength Measure Samples Initiates measurement of samples Max X Enter a maximum x value to manually rescale the graph Kinetics Scanning Max Y Enter a maximum y value to manually rescale the graph Kinetics Scanning Min X Enter a minimum x value to manually rescale the graph Kinetics Scanning Enter a minimum y value to manually rescale the graph Kinetics Scanning Molarity of cation Enters molarity of Na in incubation mixture Tm calculation in Oligos tests Next Cursor Selects a cursor point in functions using more than one cursor setting Scan Area and Scan 3 Pt Net calculations in graph Scanning Number of bases Enters number of bases in the oligonucleotide Oligos tests Number of Matched C
106. ram before you can access Run Test or Measure Samples a Setting up the test L Highlight the name of the parameter you want to parameters set 2 When the parameters are set press Save Test to save the test or Run Test to measure the blank and unknowns 144 Evolution 60 User s Guide Thermo Scientific Measuring the sample Measuring unknowns automatically using Manual 6 or Single Cell Positioner Measuring standards manually using Manual 6 or Single Cell Positioner Thermo Scientific Place the blank and unknowns in the correct cell positions Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the absorbance and concentration of each unknown Press Run Test Place the blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually When the instrument has measured all the unknowns it displays the absorbance and conc
107. rameter to either Auto 3 when using three large cell holders or Auto 6 when using six small cell holders 2 Highlight Number of Matched Cuvettes and press Enter 3 Specify the number of cells you are matching Press Enter Cell Correction 3 12pm 24Jan01 orrection Mode Discrete nms Date Corrected oC Sample Positioner Auto 6 umber of Matched Cuvettes 1 Entry Enter Number of Samples 1 5 4 Press Set nms to select the wavelengths for which the Cell Correction program will be run A list of wavelengths appears Thermo Scientific Cell Correction 15 33 26Jun02 BLANK nm 1 600 2 540 3 450 Es Entry Enter Wavelength 190 1100 a aa ae Note Cells should be matched at all analytical wavelengths because matching at one wavelength does not guarantee matching at others a 5 Highlight the position where you want to enter the first wavelength 6 Press Add nm 7 Enter the value for the wavelength and press Enter 8 Continue until you have entered all the wavelengths After all the wavelengths have been entered press Run Corr to start the Cell Correction program The Cell Correction program will measure the other cells against the blank and will record store and date the measurements From these measurements the Cell Correction program establishes the required correction factors which then are automatically applied during all subsequent tests if Cell Correction is act
108. rates a different frequency therefore if your bath operates at the resonant frequency of a cell the cell will break If a cell was cleaned in an ultrasonic bath the warranty is void by the manufacturer a Do not dry cells in an oven a Micro flowcells can be kept clean by e Flushing well with a solvent after use Thermo Scientific Cleaning the windows of the sample compartment Changing the fuse Thermo Scientific Notice Notice e Aspirating dilute acid base non filming detergent or Clorox through the cell in short bursts e Storing with distilled water in the cell Follow these guidelines to clean the windows of the sample compartment Do not use acetone or abrasive materials to clean the windows of the sample compartment Instead use a non abrasive laboratory cleaning solution Such as a commercial cell cleaning solution distilled water or alcohol e Use the liquid and a soft lint free cloth to clean the windows Do not apply too much pressure or the surface of the windows may be damaged Be sure to remove all fingerprints The fuse is located in the power entry module located at the center of the back panel of the instrument e 120VAC 2 5A Slo Blo e 240VAC 1 25A Slo Blo 2 required The instrument fuse must be replaced with the same type and rate a If the fuse fails repeatedly it may indicate a serious problem with the instrument Contact technical support as soon as possible a 1
109. s Setting baseline expiration time Thermo Scientific Utility 3 45am 16Mar07 Set Date 16Mar07 Time Format Press fT or to select item to change e e Minute AM PM Any changes you make are saved automatically even during power down by battery backup a To prolong xenon lamp life your spectrophotometer was set at the factory to automatically go into standby mode after 15 minutes To change Standby Mode time 1 Highlight Standby and press Enter tility 3 46am 16Mar07 oftware Revision 1 200 Instrument Serial Number 2M7A123005 Standby Mode OFF Go into Standby Mode now Go into Standby Mode in 15 min Go into Standby Mode in 30 min Go into Standby Mode in 60 min Go into Standby Mode in 90 min Press Tf or to select Press ENTER alculator ress fT or to select 2 Highlight the length of time you want the instrument will wait before entering standby mode and press Enter If you will be performing scans on your samples you can set a time limit for which a collected baseline will be valid To set the baseline expiration time Evolution 60 User s Guide 25 1 Setting the screen contrast 1 Highlight Baseline Expiration hr min and press Enter tility 3 47am 16Mar07 Software Revision 1 200 Instrument Serial Number 2MN7A123005 Printer Internal Date Time Setup 3 47am 16Mar07 Standb Entry Enter time 00 10 24 00 0 0FF Enter the desire
110. sition 1 Press ENTER to measure ae aT a ee 1 Place the blank and unknowns in the correct cell positions 2 Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays absorbance oligo concentration in g mL and pmol uL Dligos calc factor 14 47 28Mar02 Test Name 123 Cell 1 Abs Oligos Oligos IDH 260nm pg mL pM pL 1 0 507 14 94 1 662 Page 1 Sample 1 pea Ed Sample 136 Evolution 60 User s Guide Thermo Scientific Measuring unknowns manually using Manual 6 or Single Cell Holder Protein measurements Thermo Scientific 1 Press Run Test 2 Place the blank and unknown in the correct cell positions If a 6 Position Cell Holder is installed you can place up to five samples in the cell holder 3 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 Press Measure Sample to start the measurement If a 6 Position Cell Holder is installed press the cell position buttons to re position the cell holder and measure the rest of the unknowns manually Use the following test methods to determine the concentration of protein in a given sample Protein Direct 280 Measures the absorbance of the sample at 280 nm and determines the protein co
111. sitioner 100 mm Cylindrical Long Pathlength Cell Holder Cannot use 100 mm path cells with Cell Positioner Thin Film Filter Holders Must be installed in the B 2 and 4 positions Adjustable Filter Lens Holder Must be installed in the B 2 and 4 positions Combination Systems Not applicable Must be installed in the B 2 and 4 positions 10 Evolution 60 User s Guide Thermo Scientific Cell holders and Thermo Scientific cell holder accessories Cell holder initialization Accessories This chapter briefly describes the types of sampling and system accessories that are available for your spectrophotometer Complete descriptions and operating instructions are included with the accessories These accessories may be installed or removed without the need to switch off the instrument Term Description Cell holder assembly Complete unit that installs in the Cell holder accessory sample compartment Cell holder Part that actually holds the cell in place and is installed on a baseplate Baseplate Part that holds the cell holder and is secured in the sample compartment floor The spectrophotometer is shipped with both the 6 Position Cell Holder installed at the factory and the Single Cell Holder Directions to remove and install these cell holders and other cell holder accessories are below When you press Run Test to start a measurement the instrument will display the message Calibrating and Chec
112. sresrsrserene 12 Installing the 6 Position Cell Holder and the Single Cell Holder renee ea EEEE 13 Removing the 6 Position Cell Holder and the Single Cell OIG resorts e a aA 14 Installing accessory cell holders cece essen eee ee ees 15 Installing the internal printer optional ce0ee 15 External Printers erige eicon eentos erica le tatedvece EEEa 19 Setting up the Instrument sssssssssssssrrrrirrrrrrreeeereesnsnnnnnnnnnns 20 Entering parameter values ssssssssssrssssrrrrrssesrrrrene 20 Numeric entye o aa a a pE E EATA 20 Men SEIGCEON n scersreresrar iesean ena te aeiaai 21 On Off toggle naai aa a eaaa aiaa 21 Alphanumeric entry ssssssssssssssssnrseesssnnnnressnnnnress 21 Setting utility parameterS ssssssssssrssssrrrrrresrrrerress 21 Setting the date and time ccc cece eeeeeeeeeeeeeees 22 Evolution 60 User s Guide i Evolution 60 User s Guide Selecting standby SettingS cccccecesesaeeeeeeeeseaeees 25 Setting baseline expiration tiMe sssssssssrrresssrrrsss 25 Setting the Screen CONtraSt ccece cece eens eeeeeeee ees 26 Setting up the internal printer ssssssssssssssrrrrrssssrrn 27 Setting the utility parameters for the printer 27 Managing Stored Tests o 0 ccccssscsesssesssessssstessteeatesteesteeeesteentessees 29 Software password noc ssc avius tradlesct eattach mead susdeadoede 29 Naming a testir m npr a erine A RA A 30 SAVING
113. sssrrrrrrssrrrns 125 DNA 260 280 and DNA 260 230 cccceeeeeee ees 127 DNA with Scan 260 280 and DNA with Scan 260 230 EE E E E E T 129 Collecting a baseline SCAN sssssssrsrnsrnsnnresnrrren 131 Measuring the Sample ccccessseeeseeeeesseeeenaees 131 DNA Direct 260 dsDNA ssDNA RNA and Oligos entered factor measurements ccceeeeeeeaees 132 Oligos calculated factor cc cccccee eee eee eee neaee 135 Protein measurements ccc ce eeee cece cent eeeeeeeeeennees 137 Direct UV 280 and Direct UV 205 c cece eee 142 Warburg Christian 00 0cccecscecsecceeeeeessessccesneaees 144 Setting up the test parameters cccee cece cece eeeee 144 Cell growth vate canted str a aia animus a a su a NET 146 Setting up test paraMeters ccccccceee cece eee e ee eeee 146 Oligo Calculator vesciscssieees e tes Sides ces 148 Using the oligo calculator cececeeeeeeee ee eee eee ene eens 148 Utility Calculator FUMCtion nn escent 150 Maintenance j ccnetcenee tee ees 151 R UtINE GANG cironi ol cust cate eeeame tas DAAA EE iea eet 151 Cleaning and maintaining CellS cccceeeeeeeeeee ees 152 Cleaning the windows of the sample compartment 155 Changing THE MISE w 6c0ste sd usthinledecou teint aaa ari 155 Evolution 60 User s Guide v vi Evolution 60 User s Guide Parameters u ceececsccsssssssesescessccessscesssescassscesacessaesecaeaceesaeensaesseneasensaseneaeaeees 15
114. st measures the amount of stray light at selected wavelengths and compares them to expected values The wavelengths and expected values are preset and cannot be changed Running the stray light test takes approximately 30 seconds When running the stray light test remember that You will need Stray Light standards that are designed to measure stray light at 198 nm 220 nm 340 nm i e must have lt 0 1 T at the wavelength of interest Position B should be empty Position 1 should be used for the 198 nm stray light standard SRE 198 or equivalent Position 2 should be used for the 220 nm stray light standard SRE 220 or equivalent Position 3 should be used for the 340 nm stray light standard SRE 340 or equivalent Highlight Stray Light Press Enter Thermo Scientific Running the test Internal Printer test Thermo Scientific With the Stray Light screen displayed make sure the wavelengths and tolerances are set correctly 1 Press Start Test to run the test The results of the test appear on the screen indicating pass or fail for each wavelength If the test fails follow these guidelines e Repeat the test twice to verify that the test is failing consistently e Make sure all the filters being used are specifically designed to measure stray light at the specific wavelengths e Verify that the filters are placed in the correct cell positions e Contact technical support for more troubleshooting
115. start measurement Not available in Kinetics and Scanning Auto 3 turret auto moved B 2 4 always zeros on position B then goes to position 2 to start measurement Not available in Kinetics and Scanning Auto 6 turret auto moved B 1 2 3 4 5 always zeros on position B then goes to position 1 to start measurement Sample Volume Enters total volume of sample under Dilution Multiplier in some Bio Tests Save Test Saves all parameters of current test in internal memory for later recall all tests 164 Evolution 60 User s Guide Thermo Scientific Parameter Description Scan speed Selects speed nm min for a scan Slow Medium Fast Scanning tests only Screen Contrast Allows you to improve visibility of display by changing contrast between background and text Utility Select Test Tags highlighted test name with gt to include test in the SmartStart menu Utility Stored Tests Directory Set Max X Set cursor position in graph as the minimum X and the maximum X values for recalculating rate Set nms Allows you to enter and edit wavelength and factor values Multiwavelength tests Set Options Select factor entry or the baseline to calculate area under the peak in graph Scanning Setup correction Initiates the procedure to collect the data necessary to correct for absorbance differences between cuvettes all test types Slope Enter AAbs ACon
116. t the standard you want to delete and press Edit Standards 2 Highlight Delete Standard and press Enter to delete the standard To clear measurements L With the standard curve displayed on your screen press Edit Standards 2 Highlight Clear Measurements and press Enter All the absorbance measurements will be removed from the screen To reset standards L With the standard curve displayed on your screen press Edit Standards 2 Highlight Reset Standards and press Enter All the standards and measurements will be removed To select a different curve fit for a standard curve Note To change the curve fit for a standard curve you must display the standard curve as a graph not as a table a 1 With the standard curve you want to edit displayed as a graph on your screen press Change Fit Thermo Scientific Evolution 60 User s Guide 67 68 Evolution 60 User s Guide Standard Curve AMMONIA Test Name 3 0 1 urve Fit lope Intercept ppm i 0 538 Linear 0 6091 Std Dev 0 011 0 01272 Corr Coeff 0 995 16 21 4Feb02 BLANK Linear Through Zero Linear Regr 2nd Order Linear Regr 3rd Order Linear Regr 4th Order Segmented Press f or to select Press ENTER ESS Highlight the curve fit you want to use for the standard curve and press Enter The instrument applies the selected curve fit to the data and displays the new fit on the graph Thermo Scientific Absorbance Ratio When y
117. ter e Parallel port to an HP PCL printer To ensure that the spectrophotometer can output information correctly to the printer you need to select the appropriate device 1 Press Utility 2 Highlight Printer and press Enter Thermo Scientific Evolution 60 User s Guide 27 Internal RS232 Parallel Press Tt or to select Press ENTER to change Press ESC to exit 3 Select the printer that you want to use and press Enter until On appears 4 Press Esc to save the settings and return to the Utility screen Note Text and graphics can be output via the internal printer and an external printer on the parallel port Only text no graphics can be output via the RS232 a 28 Evolution 60 User s Guide Thermo Scientific Managing Stored Tests The Evolution 60 spectrophotometer uses test files that contain the values for all the parameters needed to runa test including the alignment of the cell changer and the other parameters for the accessories installed Once you select the values for the parameters you can assign a test name and save the test You can then restore the test and run it without having to set up the parameters again When you power down the instrument the current test is maintained by battery back up This means that when you turn the instrument on again the cell holder alignment and values for all parameters will be the same as they were when the instrument was last used When you load a test that h
118. test allows you to verify that the RS232C port is functional To run the test you will need to have the RS232 Test Plug accessory attached to the RS232C port on the back panel of the instrument Running the RS232C test takes approximately five minutes 1 Verify that the RS232 Test Plug is securely attached to the RS232C port on the back panel of the instrument With the Performance Verification screen displayed highlight RS232C Test Press Enter erformance Verification 16 59 2Mar07 8232C Test Install serial port wrap plug ress Start Test Press Start Test to run the test When the test is complete the message RS232 OK or RS232 Failed appears on the screen While the test is running you can press Stop Test to stop it If the test fails follow these guidelines Make sure that the test plug is installed correctly and is not loose or damaged Contact technical support for more troubleshooting advice Evolution 60 User s Guide 121 Table of parameters Thermo Scientific Bio Tests The Evolution 60 instrument allows you to run a variety of popular assays frequently used to characterize samples in the life science laboratory These tests fall into the following categories e Nucleic acid measurements e Protein measurements e Cell growth analysis e Oligonucleotide calculations All of the parameters for the Bio Tests described in this chapter are set at the factory If you want to change
119. the parameters specify a different name for the test and save the new test parameters To get started press Test When the Test Types screen appears highlight Bio Tests and press Enter The list of Bio Test categories appears Bio Tests 16 14 21Feb07 ucleic Acid Tests Protein Tests Cell Growth Oligo Calculator Press f or to select Start Tests Tests ATC Parameters used for each test and the literature reference for each test is located in the Parameters chapter Evolution 60 User s Guide 123 SmartStart feature Setting up a single test SmartStart 124 Evolution 60 User s Guide Note You can use this list as a reference when you are setting up tests The SmartStart feature enables you to select the test methods you use most frequently and have them appear when you start up your instrument If your laboratory runs only a single test use the SmartStart feature to select it and it will appear each time you start up your instrument Similarly if you have a set of tests you run you can use SmartStart to select them so the list appears when you start up the instrument 1 With the Bio Tests screen displayed press Stored Tests A list of all the tests on the instrument appears on the screen 2 Scroll down through the list until the appropriate test is highlighted 3 When the appropriate test is highlighted press Select Test to add the selected test to the SmartStart menu An arrow s
120. the 6 Position Cell Holder installed place the unknowns in the cell positions and press the corresponding cell position button to move the cell holder to the measuring position The absorbance ABS or transmittance T measurement appears on the display If you have the Single Cell Holder installed remove the blank and place the unknown in the cell holder The absorbance or Transmittance measurement appears on the display Measuring concentration is similar to measuring Absorbance or T Change Mode can be used to switch to concentration measurements The spectrophotometer allows you to measure concentration using either a factor or one standard to convert absorbance readings to concentration units e When you use a factor you need to specify the factor and the concentration units e When you use a standard you need to specify the concentration of the standard and measure its absorbance When Basic ATC is set to Conc Std or Conc Factor you can perform these tasks e Set the wavelength e Measure a blank e Measure a standard or enter a factor e Measure unknowns The steps for taking measurements in the two modes are similar the only difference will be whether you measure a standard or enter a factor Thermo Scientific Setting the wavelength and mode Measuring a blank Thermo Scientific L pm m Press Set nm or any other number key to set the wavelength Enter the wavelength where you want the me
121. tific Thermo Scientific Advanced A T C Abs amp T When you use the Advanced A T C program for Absorbance or Transmittance measurements you can perform these tasks e Select the measurement mode you want to use Absorbance or Transmittance e Run Cell Correction program e Recall a test or set up new test parameters e Measure a blank e Measure unknowns To get started press Test When the Test Types screen appears highlight Advanced A T C and press Enter Advanced A T C 11 42 4Feb02 BLANK Absorbance easurement Mode ave length 546 Onm Delay Time min sec 0 00 Sample Positioner Auto 6 umber of Samples 1 ore parameters Press ft or to select item to change ored i Tests Evolution 60 User s Guide 53 Recalling atest 1 From the Advanced A T C screen press Stored Tests 2 Highlight the name of the test you want to recall and press Enter From this screen you can e Set up test parameters e Run Cell Correction program e Save atest e View the list of stored tests e Measure a blank and unknowns Setting up test 1 Highlight the name of the parameter you want to parameters set Some parameters appear only if you select one of the concentration modes while others appear regardless of the measurement mode you select See the Parameters chapter for a complete list 2 When the parameters are set press Save Test to save the test or Measure Sample
122. tion 60 User s Guide Thermo Scientific Recalling a test Thermo Scientific Absorbance Difference When you use the Absorbance Difference program you can perform these tasks e Recall a test or set up new test parameters e Run the Cell Correction program e Measure a blank e Measure unknowns To get started press Test When the Test Types screen appears highlight Absorbance Difference and press Enter Absorbance Difference aa ot 3Jan0Z avelength 1 546 Onm avelength 2 546 Onm Ref Wavelength Correction off actor 1 1 000 actor 2 1 000 nits ppn Sample Positioner Auto 6 umber of Samples 1 L With the Absorbance Difference screen displayed press Stored Tests 2 Highlight the name of the test you want to recall and press Enter From this screen you can e Set up test parameters e Set up Cell Correction program Evolution 60 User s Guide 73 e Save atest e View the list of stored tests e Measure a blank e Measure unknowns Setting up test 1 with the Absorbance Difference screen parameters displayed highlight the name of the parameter you want to set 2 When the parameters are set press Save Test to save the test or Run Test to measure a blank or unknowns Note If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Note If Auto Save Data is ON you must enter a Data File Name before you can access Run Test or Measure
123. ucleotide based on a base sequence that you enter manually This concentration factor is used to calculate the concentration of oligonucleotides in your sample from the absorbance measurement Refer to the Parameters chapter for a description of the parameters and the Calculations for Oligo Calculator chapter for the default values To get started highlight Nucleic Acid Tests and press Enter A list of nucleic acid test appears Highlight the desired test and press Enter The Oligos calc factor parameter screen appears Note If Cell Correction is ON you must run the Setup Correction program before you can access Run Test or Measure Samples a Dligos calc factor 13 02 28Mar02 BLANK 2 Dilution Multiplier calculated 1 000 Sample Positioner Auto 6 umber of Samples More parameters ase Sequence TCGTCATTGAGCAGGT TCACGTGTGCTA ase Sequence Factor 29 45 umber of bases 29 Press T or to select item to change Sequence Test Tests Test Thermo Scientific Evolution 60 User s Guide 135 Setting up parameters 1 Highlight the name of the parameter you want to set 2 When the parameters are set press Save Test to save the test or Run Test to measure the blank and unknowns Measuring unknowns j Dligos calc factor 14 16 28Mar02 Measuring unknowns fest Name Oligos calc BLANK automatically A an ta A sS 1gos 1igos using Auto 6 or Auto 3 IID 260nm pg mL pM pL Insert sample 1 into po
124. unknown Evolution 60 User s Guide 65 66 Editing a standard curve To edit the concentration of a standard To add a standard Evolution 60 User s Guide You may edit the concentration of any standard ona standard curve In addition you may change the number of standards select a different curve fit or delete points from the curve Standard Curve 11 31am 21Jan01 Test Name STANDARDCURVE Edit Concentration Add Standard Delete Clear Measurements Reset Standards Press Tf or to select Press ENTER Curve Fit Linear Slope 0 002877 Std Dev 0 017 Intercept 0 002364 Corr Coeff 0 985 age 1 of 0 Standards 1 5 ress f or to view data L With the standard curve displayed on your screen highlight the standard you want to edit and press Edit Standards Si i 2 With Edit Concentration highlighted press Enter 3 Press Edit Conc or a number key 4 Enter the concentration value in the Entry field 5 When the concentration value is correct press Enter to accept the value L With the standard curve displayed on your screen press Edit Standards 2 Highlight Add Standard 3 Enter the concentration value of the additional standard in the Entry field 4 When the concentration value is correct press Enter to accept the value Thermo Scientific 5 Press Measure Standards to remeasure all the standards To delete astandard 1 With the standard curve displayed on your screen highligh
125. unknowns Taking measurements Taking measurements L With the 3 Point Net Setup screen displayed and automatically the parameters set press Run Test Auto 6 or Auto 3 The 3 Point Net measurement screen appears 98 Evolution 60 User s Guide Thermo Scientific Taking measurements manually Thermo Scientific using Manual 6 or Single Cell Holder L B Point Net 10 25 15Feb02 Test Name BILIRUBIN BLANK Abs Abs Abs Result ID 429 0 515 0 630 0 C Insert sample 1 into position 1 Press ENTER to measure gt E Place the blank and the unknowns in the correct cell positions Press Enter to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen Press Run Test B Point Net 9 26 4Feb02 Test Name 3_PTNET Cell 1 Abs Abs Abs Result ID 630 0 660 0 680 0 pg mL ir a E Blank Place the blank and unknowns in the cell holder If a 6 Position Cell Holder is installed be sure to place the blank in the B position You can place up to five samples in the cell holder Evolution 60 User s Guide 99 3 Press Measure Blank to measure the blank If a 6 Position Cell Holder is installed it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 Press Measure Sample to measure the unknowns If
126. uvettes Enter number of cuvettes that will be run in the Correction Program maximum of 5 Thermo Scientific Evolution 60 User s Guide 163 Parameter Description Number of Samples Enter number of samples to be measured in the test Not available in Kinetics or Scanning Number of Standards Enter number of standards to be measured for standard curve Printer Selects output mode internal RS 232 Parallel Utility Printout Contrast Allows you to improve visibility of internal printer hardcopy by changing the darkness of the print Utility Protein Factor Enter factor to calculate Protein concentration DNA Bio Tests Ref Wavelength Enter internal reference wavelength value for each reported measurement measures analytical wavelength amp reference wavelength Reported measurement Abs Analytical WL Abs Reference WL Ref Wavelength Correction Turns internal zeroing on or off Not available in Scanning Multiwavelength 3 Pt Net and some Bio Tests Run Corr Initiates the collection of cuvette data for cell correction Run Standard Goes to Standards entry screen Run Test Goes to the data collection screen all tests Sample Positioner Select type of Positioner 1 Cell no movement zeros and measures sample in same position Not available in Kinetics and Scanning Manual 6 cell changer moved by buttons always zeros on position B then returns to set position to
127. volution 60 User s Guide Scanning 13 33 20Jun02 Test Name Filter BLANK 0 730 Wave length ID 1 Baseline collected 20Jun02 With your scan data displayed on the edit scale screen press Auto Scale The instrument automatically adjusts the minimum and maximum values for the X and Y axes so all the data appears on the plot Thermo Scientific To use the Cursor L With your scan data displayed on the edit scale screen press Cursor Scanning 13 36 20Jun02 Test Name Filter BLANK 9 730 500 Wavelength Abs 415 2 Use Cursor to move to the desired minimum wavelength value Press Set Min X to redraw the plot using the new minimum wavelength value 3 Repeat using Cursor and Set Max X to set the new maximum wavelength To use the Manual Scale L Press Manual Scale function The manual scale options appear Scanning 9 01 15Feb02 Test Name FILTER BLANK 2 220 Wavelength 500 0 ID 1 im collected 15Feb02 2 To set the minimum or maximum value for the X or Y axis press Min Y Max Y Min X or Max X Thermo Scientific Evolution 60 User s Guide 91 3 Enter the correct value and then press Min Y Max Y Min X or Max X to accept it The instrument redraws the plot using the minimum and maximum values you have entered Performing calculations You can modify your graph by performing calculations on onthe scan data the data From the Edit Graph screen press Math When t
128. wo ways automatically or manually When you select Auto Scale the instrument automatically scales the X and Y axes so all the data appears on the plot When you select Manual Scale you select specific minimum and maximum values for the X and Y axes Whenever you modify the scale the instrument also automatically recalculates and displays the new reaction rate and result Evolution 60 User s Guide 79 When the edit screen appears you can Auto Scale function L Manual Scale function 1 80 Evolution 60 User s Guide Use the Auto Scale function to change the scale display the new graph and recalculate the results Use the Manual Scale function to change the scale display the new graph and recalculate the results Use the cursor to select new minimum or maximum values for the X axis and recalculate the results With your kinetics data displayed on the Edit Graph screen press Auto Scale The instrument automatically adjusts the minimum and maximum values for the X and Y axes so all the data appears on the plot The instrument also recalculates the results using all the data and displays it on the screen With your kinetics data displayed on the edit screen press Manual Scale The manual scale options appear Kinetics 3 24pm 27Jan01 Test Name ENZYME BLANK 0 740 A 0 583 Time 3 00 IDH A min Result 1 0 050 22 69 Enter the appropriate minimum or maximum value for the X or Y axis and then press
129. you must enter a Data File Name before you can access Run Test or Measure Samples a Collecting a baseline scan Note If a 6 Position Cell Holder is installed be sure to place the blank in the B position The instrument always uses the B position to collect the baseline a 1 Press Run Test 2 Place the blank in the B position 3 Press Collect Baseline to collect the baseline When the instrument is finished measuring the blank the message disappears Scanning 15 30 14Feb02 Test Name Cell 1 3 000 A 0 10 400 0 Wavelength 500 0 ID 1 Seline Tabular Baseline Note To switch between tabular and graphical displays press Graph Tabular a Thermo Scientific Evolution 60 User s Guide 87 Scanning an unknown Note Note Note Viewing and manipulating scan data Evolution 60 User s Guide If a 6 Position Cell Holder is installed be sure to place the unknown in cell position 1 The instrument always uses cell position 1 to scan the unknown a 1 Press Run Test If a 6 Position Cell Holder is installed be sure to place the unknown in cell position 1 2 Press Measure Sample to measure the unknown To switch between tabular and graphical displays press Graph Tabular a You may need to use the up and down arrow keys to view all the data for the scan a Test Name Filter 0 730 Scanning 13 28 20Jun02 BLANK 5 Wave length 500 ID 1 Baseline collected 20Jun0 olle

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