6850 PRISM PC Software Operating Manual REV A/06-12
Contents SECTION 1 – Introduction ....................................................................................... 4 1.1 1.2 PC SOFTWARE DESCRIPTION .......................................................................................... 4 REQUIRED PC SPECIFICATION ........................................................................................ 4 SECTION 2 – Installation ......................................................................................... 4 2.1 2.2 2.3 2.
6.5.4 6.5.5 6.5.6 6.5.7 6.6 6.6.1 6.6.2 6.7 Spectral Points Analysis ..................................................................................................... 22 Spectrum Derivative ........................................................................................................... 22 Spectrum Smooting ............................................................................................................ 22 Remeasure (Re-plot) Spectrum ...............................................
SECTION 1 – Introduction 1.1 PC SOFTWARE DESCRIPTION The Prism PC software allows the user to fully control the functionality of the Jenway 6850 variable bandwidth, double beam UV/visible spectrophotometer. The software replicates all functions from the instrument interface and adds additional functionality, extensive post-measurement tools, unlimited results storage and allows the easy export of data to other PC software packages.
3. Select Next. 4. Enter the requested user information and select Next. 5. Choose install path, then select Next. 6. Choose a program folder, then select Next.
7. Click Finish to finish the installation. 8. Connect the PC and Spectrophotometer with the USB Cable. 2.3 INITIALISATION The Prism PC software uses a USB security key to provide a licence to enable the software to run on the installed computer. Please ensure that the USB security key is inserted into a USB port of the computer at all times, when running the application. When the Prism software is installed the application shortcut will be displayed in the Start Menu folder.
2.4 SETUP COMMUNICATION PORT Select the Spectrophotometer menu and open the Comm Port Setup option. In the Communication Hub Setup window select the RS232 port being used to connect to the instrument from the available options and set the Baud Rate to 38400. Select OK.
SECTION 3 – PRISM SOFTWARE INTERFACE 3.1 MAIN WINDOW The Main Window is made up of four main areas. 1. Menu Bar 2. Toolbar 3. Data Window 4. Status Bar The information contained in each area is explained further in the following sections. Figure 3.
3.1.
Status font Setup font of status bar Customize Display Setup Peaks Identify Spectrum Peaks Valleys Identify Spectrum Valleys Zoom Activate the Zoom function Undo Zoom function Reset Return to the default display settings Search Search peak/valley one by one Connect to Connect to the instrument Spectrophotometer Re-Initialise Restart the instrument Spectrophotometer Stop measurement View dark Current Stop current measurement Refresh and display system dark current Spectrophotometer Set
Set Unit Select concentration unit Turn on/off W lamp Turn on/off Tungsten lamp Turn on/off D2 lamp Turn on/off Deuterium lamp D2/W Switch Point Set switch point of Deuterium and Tungsten lamps Auto-sample Comm.
Divide Divide one spectrum from another Savitzky-Golay Smooth a spectrum with the Smoothing Filter Savitzky-Golay method filter Derivate Display the derivative of a spectrum Remeasure Remeasure a spectrum New Window Cascade Opens a new measurement window the same as currently displayed Multiple windows are displayed in a cascade on screen Window Help Tile Multi windows are tiled on screen Arrange Icons Arrange all icons minimized Split Split display area Help Topics Displays the Help i
SECTION 4 – PHOTOMETRIC MEASUREMENT 4.1 WAVELENGTH SELECTION A single wavelength photometric measurement is made by firstly selecting the Goto Wavelength icon from the Toolbar. A window will open that allows the selected wavelength to be entered. Once the wavelength has been entered, select the Goto button. 4.2 SAMPLE MEASUREMENT If required, select the Zero button to reset the Readout value to 0 Abs or 100%T, then insert the blank and sample cuvettes into the sample chamber of the instrument.
SECTION 5 – MULTIWAVELENGTH / QUANTITATION The multi wavelength and quantitation measurement mode enables measurements of absorbance, % transmittance and concentration to be performed. In this measurement mode it is possible to perform a photometric measurement at up to 20 separate wavelengths. This mode also allows the concentration of an unknown sample to be determined against a calibration curve or a known concentration factor at up to three separate wavelengths. 5.
5.3 SAMPLE INFORMATION The Information tab allows the user to enter sample and standard information. Additional notes can be entered into the Memo field if required. 5.4 SAMPLE MEASUREMENT AND DISPLAY OPTIONS The Sample tab contains a result window and a Control window. The Control window contain options to Start, Delete, Modify, Recalculate, change the Data Font and Print. 5.4.1 Control Window – Start Select the Start button to initiate a multiwavelength measurement.
5.4.4 Control Window – Recalculate Select the Recalculate button to re-evaluate the concentration result when the Quantitation mode is enabled. 5.4.5 Control Window – Data Font Select the Data Font button to format the result data. 5.4.6 Control Window – Print Select the Print button to generate a report of the recorded data. The user will be asked to confirm if the report should include the text entered into the Information tab fields and the data contained in the result window. 5.
5.5.1.2 Entering Known Calibration Curve Constants The Use Standard Samples checkbox should be clear and the Curve Fit option should be set to the appropriate option. The known calibration curve constants should then be entered into the appropriate K0 – K3 fields where:Concentration = (K3*X3)+ (K2*X2)+ (K1*X) + K0 X = Photometric value of the sample 5.5.
Once the standard measurement is complete and the name and concentration information has been entered, select OK to transfer the information to the main standard tab. Repeat this procedure for each standard sample. 5.5.2.2 Calibration Curve Settings and Display The calibration curve display settings are edited by selecting the Display Setting tab. The calibration curve’s labels, axis range and interval values and concentration units can be edited in this tab.
5.5.3 Sample Measurement To quantify an unknown sample select the Sample tab and select the Start button. A new window will open allowing the sample name and final reported photometric reading to be edited. The actual photometric reading of the standard will be displayed in the Readout window and this value will default to the deltaAbs field. Select OK to transfer the data to the sample result table.
SECTION 6 – SPECTRUM The spectrum measurement mode enables measurements of absorbance or % transmittance over a range of wavelengths to be performed. The absorbance or % transmittance at each wavelength is plotted graphically. Post measurement tools such as peaks/valleys, derivatives and spectral points analysis can be performed. This operating mode can be used to partially characterise a sample. 6.1 SPECTRUM MODE SCREEN Select the Spectrum Measurement icon on the toolbar.
6.4 SAMPLE MEASUREMENTS Insert a cuvette containing the blank solution into the reference position in the sample chamber and insert the cuvette containing the sample solution into the sample position. Close the instrument lid and measurement icon measurement is select the Start a on the toolbar. Once the complete the measured spectrum scan will be displayed on the screen. The scan can be cancelled by selecting the Stop a measurement icon 6.5 POST MEASUREMENT TOOLS 6.5.
6.5.3 Spectrum Zoom Function Select the Zoom function icon from the toolbar. Position the cursor in the upper-left corner of the area you want to select. Hold the left mouse button to drag the cursor to outline the spectrum area you want to enlarge. Release the mouse button. The part of the spectrum which is displayed within the outlined area will be enlarged. Select the undo zoom icon to restore the previous view settings. Select the Zoom function icon again to exit the zoom function. 6.5.
6.5.7 Remeasure (Re-plot) Spectrum Click on the toolbar. A dialogue box appears asking the user to specify the frequency of the data points in the re-plotted spectrum. Select OK to confirm. The re-plotted spectrum will be displayed. 6.6 OVERLAY SPECTRA The Prism Pc software can display multiple spectrum scans simultaneously on the screen by either measuring additional samples or loading stored data. The active spectrum is selected from the drop down menu on the toolbar.
6.6.2 Delete Displayed Spectrum Select the spectrum to be deleted in the active spectrum drop down box (see 6.6). Click on the toolbar to remove the spectrum from the display. 6.7 SPECTRUM DISPLAY AND PRINT OPTIONS The display and print options for the selected spectrum can be accessed by selecting the icon on the toolbar. The display range, peak and valley labels, axis legends, spectrum title, display colours, print information options and printout notes can be edited in the dialogue box that appears.
SECTION 7 – KINETICS The kinetics measurement mode enables the absorbance or % transmittance of an active molecule to be measured over a period of time; for example enzyme analysis of horseradish peroxidase. The absorbance or % transmittance is measured at regular time intervals at a set wavelength over a period of time. The results are plotted on a graph to show the change in absorbance or % transmittance over time.
7.4 SAMPLE MEASUREMENTS Insert a cuvette containing the blank solution into the reference position in the sample chamber and insert the cuvette containing the sample solution into the sample position. Close the instrument lid and measurement icon measurement is select the Start a on the toolbar. Once the complete the measured spectrum scan will be displayed on the screen. The scan can be cancelled by selecting the Stop a measurement icon 7.5 POST MEASUREMENT TOOLS 7.5.
7.5.3 Spectral Points Analysis Select the Scan spectrum icon from the toolbar. Move the cursor over the kinetics spectrum to trace the scan and display the scan data. Select the Scan spectrum icon again to exit the spectral points analysis mode. 7.5.4 Kinetics Derivative Click on the toolbar. A dialogue box appears allowing the user to select the derivative function required (1-10) and enter a name for the calculated derivative spectrum. Select OK to confirm.
7.6.1 Kinetics Display and Print Options The display and print options for the selected scan can be accessed by selecting the icon on the toolbar. The display range, peak and valley labels, axis legends, spectrum title, display colours, print information options and printout notes can be edited in the dialogue box that appears. 7.6.2 Calculate Rate of Change The rate of change for the selected kinetics scan is calculated by selecting the Display and Print settings icon on the toolbar.
SECTION 8 – DNA/PROTEIN The DNA/Protein measurement mode allows the user to measure multi-wavelength absorbance ratios, such as 260nm/280nm and 260nm/230nm, which are commonly used to estimate a protein or nucleic acid sample’s purity. The mode also includes calculations that can be used to estimate the concentration of the protein or nucleic acid sample. Four commonly used protein assay methods are pre-loaded in this measurement mode.
8.3 DETERMINATION OF NUCLEIC ACID CONCENTRATION Select the required method parameters as in 8.2. 8.3.1 Sample Measurements To quantify an unknown sample select the Sample tab. Insert the sample cuvette into the sample cuvette holder and the blank solution into the reference sample holder. Select the Start button. A new window will open allowing the sample name to be edited.
8.4.2 Constructing a New Calibration Curve 8.4.2.1 Measuring Standard Samples Select the Standard tab. Insert the first standard into the sample position in the sample chamber and the reference into the reference position in the sample chamber. Select the Start button to initiate the measurement A new window will open allowing the Standard name and concentration to be edited. The actual photometric reading of the standard will be displayed in the Readout window.
A new window will open allowing the sample name to be edited. The actual photometric reading of the standard will be displayed in the Readout window and this value will default to the C-DNA field. Select OK to transfer the data to the sample result table. The sample result table will display the measured absorbance value concentration value.
SECTION 9 – APPENDIX The multicell accessory measurement mode allows the user to measure a sample’s photometric absorbance or %transmittance at up to 10 wavelengths. Following sample measurement, the photometric readings are displayed on the multi-wavelength mode screen. 9.1 CALCULATIONS IN QUANTITATION MODE Single Wavelength Method ：Abs.=A1 Double Wavelengths Method ：Abs.=m*A1-n*A2 Three Wavelengths Method ：Abs.=A1-(WL1-WL2)*(A2-A3)/(WL2-WL3)-A3 9.
SECTION 10 – TECHNICAL SUPPORT 10.1 TECHNICAL SUPPORT Jenway have a dedicated Technical Support team made up of experienced scientists who are on hand to help with any applications advice and questions you may have about our products and how to use them. If you require any technical or application assistance please contact the team at: E-mail: jenwayhelp@bibby-scientific.com.
