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世聯(lián)翻譯公司完成FORAM軟件說明英文翻譯
發(fā)布時(shí)間:2018-01-15 08:48 點(diǎn)擊:
世聯(lián)翻譯公司完成FORAM軟件說明英文翻譯
CONTENTS1. Turning on the FORAM 12. Logging in to the FORAM Software 2a. Using the Research (Non-Casework) Mode 2b. Creating a new case 3c. Opening an existing case 5d. Creating a New Operator 6e. Deleting Operators 7f. Changing Save Location 83. Setting up the FORAM 9a. Objective Lens Selection 9b. Sample Placement and Stage Manipulation 10c. Using the Light Shields 11d. Laser Power Control 124. Calibrating the FORAM 13a. Automatic Calibration 13b. Manual Calibration 165. Recording Spectra and Image Capture 19a. Recording and Saving Individual Spectra 19b. Recording group spectra 23c. Saving Spectra as a Group 25d. Recording a Sample Image 266. Spectral Display Options 277. Background Correction 33a. Automatic Background Correction 33b. Manual Background Correction 358. Spectral Comparison Options 37a. Overlay of Spectra 37b. Offset of Spectra 38c. Vertical Scale 39d. Horizontal Zoom 40e. Marker 41f. Labels 42g. Zoom Window 449. Exporting Data and Printing Results 4510. Database Searching 47a. Searching Against a Database/Library 47b. Creating a New Database/Library 49c. Adding a Spectrum to a Database/Library 50d. Deleting Spectra from a Database/Library 52e. Deleting a Database/Library 54f. Adding a Commercial Library or Database 56Appendix I – Hints/Tips for Obtaining Good Spectra 59Appendix II – Application of SERRS 62Appendix III – Routine Quality Assurance Checks 63Appendix IV – Technical Information on Laser Spot Sizes and Fields of View 65Appendix V – Application Notes 671. Turning on the FORAMThe FORAM should be switched on in the following order to ensure correct communication between the instrument modules.1. Switch on the computer and wait for the Windows desktop to load.2. Switch on the FORAM (switch at rear of module).3. Start the FORAM software using the shortcut icon on the PC Desktop.4. The FORAM log-in screen should now be displayed5. The FORAM should be allowed to warm up for at least 10 minutes before use for optimum performance.2. Logging in to the FORAM SoftwareWhen the software opens the user must choose whether to create a new case, open an existing case or work in a research (or non-casework) mode.a. Using the Research (Non-Casework) Mode1. At “Case Selection” select Close. There will be no case-details or calibration log for this selection.b. Creating a new case1. Check that the Operator field is correct. Depending on the software set-up this may be ‘Default’ or the user’s name.2. At “Case Selection” choose select New Case.3. The user must then enter the case details.4. The Case and Operator 1 fields are mandatory. Operator 1 is automatically populated with the Operator in use at “Case Selection”.5. Once all the relevant case details have been entered select OK.6. If New Case was selected by mistake select Cancel.7. Once a New Case has been created the user will be reminded to calibrate the FORAM. Select OK.8. This reminder will appear after every new case is created. If it is not required select the tick-box stating: Do not show this message again.Viewing and Editing Case Details1. Case Details can be viewed and edited after case creation.2. Select Case Details to view the case details.3. To edit the details select Edit.4. To complete the process select Save, which will have replaced the Edit button during editing.c. Opening an existing case1. Check that the Operator field is correct. Depending on the software set-up this may be ‘Default’ or the user’s name.2. At “Case Selection” select the required case so that it is highlighted in blue.3. Select Open Case.d. Creating a New Operator1. Select Edit at “Case Selection”.2. To create a new operator select New Operator.3. Enter an operator name in the Operator field.To choose an alternative save location choose Edit next to the Text Path field and either specify a new field by choosing a folder to save to or choose Make New Folder to create a new save location. When finished select OK.e. Deleting Operators1. Select Edit at “Case Selection”.2. To delete an operator select the operator to be removed so that the operator name is highlighted in blue. Then select Delete Operator.3. For the action to be completed select OK for confirmation.f. Changing Save Location1. Select Edit at “Case Selection”.2. To change save location select the desired operator so that the operator name is highlighted in blue. Then select Edit Path.3. Specify a new field by choosing a folder to save to or choose Make New Folder to create a new save location. When finished select OK.3. Setting up the FORAMThe following instructions may all be performed whilst the FORAM is warming up.a. Objective Lens Selection1. The FORAM has three objective lens: 5x, 10x and 20x. The objective lens in use facing directly downwards.2. To change between objective lenses rotate the turret to the desired lens.Lens Magnification Colourx5 Redx10 Yellowx20 GreenThe x5 objective lens is used for positioning the sample whereas the x10 and x20 objective lenses are used for recording spectra. The x10 objective lens allows the user to collect a spectrum from a larger area whereas the x20 objective lens allows the user to be more specific with the area that they are recording a spectrum from.b. Sample Placement and Stage Manipulation1. Place the sample directly under the laser spot.2. Focus on the sample using the Z control of the manipulation stage.3. Fine position the sample using the X and Y controls of the manipulation stage.c. Using the Light ShieldsThe FORAM is sensitive to ambient light. To achieve best results ambient light should be blocked. This can be achieved by using the light shields on the objective lenses.1. Unscrew the shield attached to the objective lens in a clock-wise motion until the shield is lightly touching the sample.d. Laser Power ControlThe power of the laser is one variable that can affect the Raman signal that is measured by the FORAM.1. The laser power is adjusted manually by the control on top of the FORAM as required.When examining dark or colored samples it is recommended to take measurements at 10% laser power due to the sample potentially melting or photo-degrading at the 100% laser power setting.4. Calibrating the FORAMThe FORAM is calibrated during production. For regular calibration checks, the standard Polystyrene beads supplied with the instrument should be used. Polystyrene produces a Raman spectrum with peaks at characteristic wavelengths. Polystyrene was selected as the calibration standard in accordance with the ASTM International Standard E 1840 – 96 Raman Shift Standards for Spectrometer Calibration.The FORAM should be calibrated daily before use.Each time a new case is created there will also be a prompt to calibrate the case for case integrity. An automatic calibration carried out when a case is open will have its information added to a calibration log.a. Automatic Calibration1. Position a polystyrene bead directly under the laser.2. Focus on the polystyrene bead and fine tune the position using the XYZ translation stage. The surface of the polystyrene bead should be clearly seen in the live video.3. Set the scan time to 1 second average count to 6.4. Record a spectrum of the polystyrene bead.5. Navigate to the Calibration menu and select Auto Calibration.6. Select Calibrate.7. If the calibration is successful a message will be shown in green stating that the calibration has been completed.8. If the calibration is not a message will be shown in red stating that the calibration was not completed and that a manual calibration will need to be carried out.9. Select Close to complete the automatic calibration process.b. Manual Calibration1. Position a polystyrene bead directly under the laser.2. Focus on the polystyrene bead and fine tune the position using the XYZ translation stage. The surface of the polystyrene bead should be clearly seen in the live video.3. Set the scan time to 1 second average count to 6.4. Record a spectrum of the polystyrene bead.5. Navigate to the Calibration menu and select Manual Calibration.6. Select the peaks as highlighted in the Manual Calibration window. This will populate the fields for peak number and position.Starting manual calibrationAll peaks selected7. If a mistake was made during peak selection, select Reset.8. Following peak selection select Calibrate.9. Select Confirm to confirm the manual calibration.10. Select Close to complete the manual calibration process.Please note that during Manual Calibration peaks must be selected in the correct order. Failure to do so will not allow the system to calibrate.5. Recording Spectra and Image Capturea. Recording and Saving Individual Spectra1. Place the sample under the objective lens and bring the object image in to focus using the XYZ translation stage.2. Enter the number of Average Counts for the sample being analysed. Increasing the Average Count improves the signal-to-noise ratio but also increases the data acquisition time.Scan Time (s) Minimum Recommended Average Count> 1 60.01 – 1 123. Select the Auto Exposure tick box. Auto Exposure will automatically determine the optimum scan time for the sample being analysed.4. Select the Background Correct tick box. Background correct will automatically background correct the spectrum being recorded to allow for better spectra comparison.5. Record the spectrum of the target object using by selecting the Record Spectrum button.6. The spectrum of the target object is displayed on the spectral graph to the right of the screen; the image updates as each new count is acquired.7. When the spectrum acquisition is finished, the spectrum details appear under the Main tab.8. Select the Spectrum Title field and enter an appropriate file name in accordance with your local laboratory procedure.9. Press the Return key on the keyboard to complete the renaming process.10. Select Save Spectrum and store in an appropriate location on the computer.11. Repeat steps 1-9 to record the spectrum of any further samples in the case.12. Each new spectrum is shown on the spectral graph in a different colour and the same colour is used to identify the record in the Main tab table.b. Recording group spectraTo save time entering sample details for each individual measurements, it is possible to Create a Group for samples from a specific source.1. Before recording any spectra, select the Create Group tool button.2. Enter the required Group name into the pop up window displayed.3. Record the transmission spectrum of the sample using the method detailed in Steps 1 – 9 of Recording and Saving Individual Spectra (Focus on target object => Record Spectrum).4. Each new spectrum is shown on the spectral graph using the same colour.and is identified by the Group name and a sequentially numbered value (1, 2, 3...., etc) under the Main tab.5. Highlight the relevant spectrum by selecting it using the left mouse button so that the spectrum is highlighted in blue. Select Save Spectrum and store the record in an appropriate location on the computer in accordance with your local laboratory procedure6. The Group name will be assigned to each new sample analysed until the Create Group function is used to set up a new group or the FORAM software is closed. If a sample from a different source is accidently analysed whilst the Group function is active, it’s possible to save the record using the correct identifier by selecting the relevant Spectrum Title and entering the appropriate details.c. Saving Spectra as a GroupThe Save Group function is used to save multiple spectra to a single data file to speed up data retrieval for analysis at a later date. The function can be applied to spectra previously stored as individual records or to multiple records that have yet to be saved. The data in each individual spectrum within a Saved Group (.FORAM_group) remains unchanged and is not averaged or otherwise altered by use of this function.1. Select the records from the Main tab table to be stored as a Group. This is achieved by using Select All from the menu options available from right clicking on the Main tab tableor by using the cursor and the standard Microsoft features using the <CTRL> key to select individual records or the <SHIFT> key to select a run of records.2. Select Save Group to store the selected records in an appropriate location on the computer in accordance with your local laboratory procedures.d. Recording a Sample ImageIn addition to saving the spectrum of an analysed sample, the FORAM software also enables the operator to save the image of the analysed sample seen on the Live Video screen. Select the Save Image tool button below the Live Video screen.The saved image shows the sample from which the measurement was obtained without measurement area cross-hairs superimposed.6. Spectral Display OptionsLoading Spectra for AnalysisPreviously stored spectra are opened using the Open command from the File menu or using the shortcut tool button on the toolbar.Changing Spectrum Display ColourThis option avoids confusion arising due to spectra from different sources have been assigned the same default display colour when originally recorded.1. To change a spectrum’s colour, select the required spectrum from the Main tab menu and either Change Colour of Selected Spectrum from the Main tab right-click menu or using the Change Colour tool button.2. Select a suitable colour from the pop up window shown and select OK.3. The selected spectrum’s colour is updated on the spectral graph and in the Main tab table.a) Original Display Colours b) New Display Coloursa) Original display colour b) New display colourGroup Selected SpectraThis option enables the spectra from different samples from the same source to be grouped together. Grouped spectra are shown on the screen the same colour and treated by the FORAM software as coming from a single source.1. Select the records from the Main tab table to be Grouped using the Select All option from Main tab mouse menu or by using the cursor and the standard Microsoft features using the <CTRL> key to select individual records or the <SHIFT> key to select a run of record.2. Right click on the Main tab table again and select Group Selected Spectra.3. The following message is shown:4. Select the Group Colour tool button, pick a suitable colour from the pop up window shown and select OK.5. The chosen colour will appear in the Group Spectra window. Select OK to confirm or use the Group Colour tool button to make a different selection.6. The selected spectra and associated records in the Main tab table will also be shown in the same colour.Displaying Averaged SpectraThis option generates an averaged spectrum from two or more selected spectra. The function does not alter any of the original data files used in the averaging calculation. If comparisons are carried out using averaged spectra it is recommended that the original data files used to generate the average spectrum are saved so that the source data is available for review if required.1. Select the records to be averaged from the Main tab table.2. Select the Display Average of Selected Spectra option from the Main tab right-click menu.3. The Averaged Spectrum is displayed in a new colour on the spectral graph and added to the records listed in the Main tab table.Original data Averaged spectrum shown in redGridlinesGridlines are automatically shown on the spectral graph. To removes these lines from the display and printed report, deselect the Show Gridlines option on the View menu.The colour and appearance of the Gridlines may also be changed to suit individual preferences using the Set Up option from the System menu.Plot ThicknessThis option allows users to change the thickness line used on the spectral graph for the selected spectrum or spectra. It may be used to highlight a specific spectrum where spectra are overlapping and not clearly distinguishable by colour or where use of the Offset function is not desired.1. Select the spectrum or spectra from the Main tab table.2. Right click on the Main tab table again and select the Plot Thickness required from the menu shown.3. The graphical screen is updated to show the new spectrum.Smoothing spectraAlthough the Average Count function will smooth spectra by removing noise there is a supplementary software function for smoothing spectra.Within the FORAM software the smoothing is carried out using a 5-point Savitsky-Golay filter.To smooth spectra select the Smooth tickbox in the Main tab Spectra Details window7. Background CorrectionFor easier spectral comparisons it is recommended that spectra are background corrected. Background correction allows for spectra to be overlaid, spectra to be offset in a more presentable style and finally smaller peaks will become more visible as a result of removing the unused area beneath the spectrum baseline.Background correction can be carried out automatically and manually.a. Automatic Background Correction1. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the Analysis tab.3. Select the spectrum to be background corrected by selecting the corresponding tick box.4. Automatic Background Correction is selected by default.5. Select the Automatically Correct Background button.The uncorrected spectrum will display with the background correction line shown. The background corrected spectrum is also shown.6. To remove a background correction select the Remove Background Correction button.7. After one spectrum has been background corrected all remaining spectra can also be corrected by selecting the Apply to All Spectra button.The Apply to All Spectra function will create the best background correction for each spectrum, rather than using the background correction created for the first spectrum and applying that to all remaining spectra.8. If more than one spectrum has been background corrected it is possible to remove all background corrections. This is done by selecting the Remove from All Spectra button.b. Manual Background Correction1. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the Analysis tab.3. Select the spectrum to be background corrected by selecting the corresponding tick box.4. Select the Manual Background Correct radio button.5. Select points on the baseline of the Uncorrected Data to start background correcting the spectrum.6. As points are added to the baseline of the Uncorrected Data the Corrected Data screen will update showing the current result of the manual background correction.a) During manual background correction b) Result of manual background correction7. To remove a background correction select the Remove Background Correction button.8. After one spectrum has been background corrected all remaining spectra can also be corrected by selecting the Apply to All Spectra button.The Apply to All Spectra function will create a background correction for the other spectra using the same point positions that the user selected. It is not recommended to use this function in Manual Background Correction.9. If more than one spectrum has been background corrected it is possible to remove all background corrections. This is done by selecting the Remove from All Spectra button.8. Spectral Comparison Optionsa. Overlay of SpectraOnce spectra have been baseline corrected the spectra will be overlaid (on top of each other). This allows for easier examination for presence and absence of peaks between spectra.b. Offset of SpectraThis option enables the vertical positions of selected spectra to be increased by a fixed amount across the full wavelength range so altering their position displayed on screen.1. Select the required spectrum or spectra from the Main tab table.2. Use the buttons to increase / decrease the Offset as required.3. The spectral graph is updated as the offset is changed.a) Original display b) Offset display4. A second spectrum may now be selected and a different Offset applied if desired.5. The Reset Offset tool button is used to remove all offsets applied.c. Vertical ScaleThe spectral graph vertical scale is automatically scaled to show the acquired spectra in full. This option enables the user to disable the Auto Scale function and manually set the vertical scale.1. Use the buttons to increase / decrease the vertical scale as required. The Auto Scale function is automatically disabled.2. To reactivate the automatic scaling, select Auto Scale check box.d. Horizontal ZoomThe Zoom function enables users to magnify the spectral graph from the full, default range from 300cm-1 to 2100cm-1 to a maximum zoom range of 450cm-1. The scroll bar at the top of the spectral graph is used to scan through the different wavelengths in the collected spectra. Unlike Zoom Window the spectra shown are not normalised when displayed.1. Move the slider on the Zoom bar to the desired magnification.2. Use the scroll bar at the top of the spectral graph to scan through the wavelength range.Please note these spectra have also been offset.e. MarkerThis option places a vertical line on the spectral graph. The marker line can be dragged to any position throughout the wavelength range shown to help users align spectral features at a given wavelength.1. Select the Marker check box in Display Options; the marker line is automatically shown at 1000cm-1.2. Use the mouse to drag the marker to any position required desired in the spectral graph; the relevant wavelength is automatically shown at the top of the screen.Please note that the marker is not printed.f. LabelsThis option provides users with a means of placing labels on the recorded spectra.1. Select the Labels check box in Display Options.2. Select the spectrum in the Spectra Details that you wish to add labels to. Once the spectrum has been selected it will be highlighted in blue.3. Using the mouse left click on the peak you want to label. The label will then appear.4. To remove a label right click with the mouse. This will remove the last added label.5. To remove all labels select the Clear Labels button.The colour and appearance of the Gridlines may also be changed to suit individual preferences using the Set Up option from the System menu.g. Zoom WindowThe Zoom Window provides users with a magnified view of a selected area of the overall wavelength range enabling more detailed comparisons to be made. The function also normalises the spectra displayed in the upper, Zoom Window making it easier to compare the significant spectral features between samples with significantly different depths of colour.1. To open the Zoom Window select Zoom Window from the View Menu.2. This opens a seconds window above the original spectrum / spectra in which only the data from the wavelengths shown between the blue and red markers are shown.3. To change the zoomed wavelength range shown, drag the blue and red markers using the mouse cursor to the new positions required.9. Exporting Data and Printing ResultsThe FORAM software provides the user with a range of options for exporting or printing the spectral data obtained during examinations. Data can be exported in text or graphical format and copied into local laboratory formats for further processing or reporting. An overview of the options available is provided below.File MenuPrintPrints the information currently displayed in the window to the right of the screen. On the Main tab screen the spectral graph will be printed; on the Analysis tab screen the selected spectrum in its uncorrected form with baseline will be printed; and on the Database tab screen the search and match spectrum will be printed.Print PreviewGenerates a preview of the information to be printed.Page Set UpEnables the user to select paper size, orientation and margin size for printed reports.Print to ClipboardCopies spectral graphs to the Windows Clipboard enabling the user to paste the results into supported applications such as Microsoft Word.Print OptionsEnables the user to determine whether the Spectrum report includes: a) the spectral graph and associated data file names or b) the spectral graph, data file names and the Scan Time and Average Counts used to acquire the data.Export All Data to Clipboard as XY PairsExports the intensity value at each wavelength from 400-2000cm-1 for all data files in the Main tab table to the Windows Clipboard to be pasted into supported applications such as Microsoft Word and Excel.The XY pairs for an individual spectrum can be exported by using the right mouse click menu on the Main tab and selecting Export Spectrum XY Pairs to Clipboard.10. Database SearchingThe FORAM software includes the functionality to create databases and search against them. In the future the FORAM software will also include the functionality to incorporate commercial libraries and search against them.Searching against databases is accessed through the Database taba. Searching Against a Database/Library1. A spectrum must be available to be used to search against a database/library. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the spectrum you wish to search by selecting it using the corresponding tick box. The selection of a spectrum can be confirmed by the spectrum name being highlighted in blue3. Select the library/database(s) that you wish to search against by selecting from the Available Libraries.4. Search against the library/database(s) by selecting the Search button.5. The results of the search are shown in the Search Results. The results are sorted from the highest % Match to the lowest % match. The spectrum name and the library/database name are also shown.6. To compare the search spectrum with other database spectra select the required spectrum name from the Search Results. The Match Spectrum will then update with the selected spectrum.7. By default only spectra with a % Match over 50% will be displayed. This threshold can be adjusted to show spectra with different % Match. The Search Threshold ranges from 1% to 100% and is adjusted by selecting the appropriate arrow buttons to increase or decrease the Search Threshold.8. The results can be removed by selecting the Clear button.The search algorithm, as shown by the Search Mode, is Pearson’s Correlation of the first derivative. This a standard algorithm used in spectral searching/matching.b. Creating a New Database/Library1. A spectrum must be available to be used to add to a database/library. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the Add Spectra to Database button.3. Select the Create New Library button.4. Enter the name for the new database/library.5. Select OK to confirm the name. Select Cancel to return to the previous window without creating a new menu.6. To complete the creation of a new database/library select Close. The Available Libraries in the Database tab will now be updated showing the new database/library.c. Adding a Spectrum to a Database/Library1. A spectrum must be available to be used to add to a database/library. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the spectrum you wish to search by left mouse clicking on its name. The selected spectrum name will be highlighted in blue. More than one spectrum can be selected by holding the left mouse button and dragging over the names to be selected. The <Shift> key and <Ctrl> can be used in conjunction with the clicking on spectra names to select multiple spectra.3. Select the Add Spectra to Database button.4. Select the spectrum/spectra to be added to a library/database.5. Select the library/database you wish to add the spectrum/spectra to by selecting the relevant tick box.6. Select the Add Selected Spectra to Selected Library button.7. To complete the process select Close.The Available Libraries in the Database tab will now be updated with the relevant database/library containing the added spectra.d. Deleting Spectra from a Database/Library1. A spectrum must be available to access the Library Editor. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the Add Spectra to Database button.3. Expand the database/library you wish to delete a spectrum from by selecting the + next to the database/library name.4. Select the spectrum you wish to delete.5. Select the Delete Selected Spectra button.6. To confirm the deletion of the spectrum click OK.7. To complete the process select Close. The Available Libraries in the Database tab will now be updated with the deleted spectra removed.e. Deleting a Database/Library1. A spectrum must be available to access the Library Editor. Either record a spectrum as detailed in Recording Spectra or open a spectrum as detailed in Spectral Display Options.2. Select the Add Spectra to Database button.3. Select the database/library you wish to delete.4. Select the Delete Selected Library button.5. To confirm the deletion of the database/library click OK.6. To confirm the deletion of the database/library click OK.Deleting a database/library is confirmed twice. The first instance confirms the deletion of the library. The second instance gives notification that the database/library contains spectra which will be deleted with the deletion of the database/library.7. To complete the process select Close. The Available Libraries in the Database tab will now be updated with the deleted database/library removed.f. Adding a Commercial Library or DatabaseThe FORAM software supports commercial libraries. Typically the commercial libraries will be purchased with the FORAM. To use the libraries they must be added to the software by the user.To add the library:1. Check that the Database tab is selected.2. Select Library Directories from the File menu.3. The Library Directory dialogue window will open4. Select Add Path5. Browse to the Library location. This can be a location on the hard disk (C:) or a temporary file location (CD/DVD Drive)6. The Library Directory dialogue window will now be updated displaying the new directory location7. To finish the process select Close.If required a path can be removed by highlighting the relevant path so that it is highlighted in blue and then select Delete Path.8. The Available Libraries within the Database Tab will now be updated to show the included library.Note: If a temporary file location (for example the CD/DVD Drive) was selected during this process when the CD/DVD is removed it will not be possible to search against the libraries on that CD/DVD until the CD/DVD is replaced in the CD/DVD Drive.Appendix I – Hints/Tips for Obtaining Good SpectraThe FORAM is a powerful tool for comparing a variety of forensic materials. To obtain results from the FORAM that enable the user to achieve the best interpretation of the findings, there are a few basic rules when obtaining sample spectra.• All samples in a case must be prepared using the same materials and preparation methods.• Select samples from each source for comparison that represent the full range of variation present.• Thin film samples can be attached to aluminium foil on a glass microscope slide to allow for better positioning and focus.• Avoid mounting transparent samples on glass slide, as glass can weakly fluoresce and mask the raman signal• Select an area of the sample that, wherever possible, is clean and free from damage. The presence of dirt and/or damage can have a significant effect on the sample spectrum obtained due to diffraction of the light passing through the sample.• Before recording spectra the sample should be placed and focused so that a circular laser spot can be seen close to the centre of the live video image.• Ensure that no ambient light is allowed through to the detector. Ambient light will cause positive/negative peaks to be observed that are not caused by the sample. Ambient light can be blocked by: Working in a dark room environment with lights switched off and using curtains to cover windows. Using the light shields attached to the objective lenses. [Place a cloth over the FORAM hardware unit]• Although 100% laser power will produce a result in the quickest time it is sometimes advantageous to use a lower laser power of 25% or 10% with a longer scan time. This allows more information to be collected by the spectrometer and provide better results. This is true for highly absorbing samples, which may burn or melt on the highest laser power setting.• Set the Average Count to a minimum of six (6). Using the Average Count function will reduce the amount of noise observed in the final result.• Obtain a minimum of six (6) measurements from each sample to observe any variation.Extraction MethodIt has been observed that results can be improved (notably for laserjet toners) by extracting the sample from the document. The method is carried out as follows:1. Take a small portion of the sample (for toner, this would be an area of toner on a piece of paper).2. Place the portion of the sample in a 350μl vial.3. Submerge the sample in acetone (typically 40μl).4. Shake the vial vigorously for a minimum of ten (10) seconds. The acetone may become discoloured if the sample was toner.5. Place a strip of aluminium foil on a microscope slide.6. Pipette the sample ‘extract’ solution from the vial using a micropipette and place a drop of the solution on the foil.7. Allow the extract solution to dry.8. Repeat Step 6 in the same position to concentrate the sample area.9. Record a minimum of six (6) spectra of different regions of the dried sample extract using the FORAM.10. Repeat the method for other samples.The final sample should appear as a white film on the foilAppendix II – Application of SERRSSERRS is the combination of two separate methods, Resonance Raman (RR) spectroscopy and Surface Enhanced Raman Scattering (SERS) spectroscopy. SERRS provides an improvement in sensitivity of up to fifteen orders of magnitude greater than standard Raman spectroscopy to be achieved. This allows fluorescence to be quenched and can cause peaks arising from some vibrational modes to be enhanced. As a result a SERRS spectrum may be different to the original Raman spectrum which then allows for further discrimination.It must be noted that not all dyes and pigments will show this effect. For example the spectrum of many black ballpoint inks, contain peaks from a copper Pthalocyanine component, with an underlying fluorescence curve from Methyl Violet dye component.When treated with colloid the Methyl Violet enhances, whilst the copper Pthalocyanine does not, giving rise to a different spectrum consisting of peaks from the copper Pthalocyanionne and SERRS peaks from the methyl violet.• All samples in a case must be prepared using the same materials and preparation methods.Method of Application1. Using the micropipette place 0.5 μl of Poly-L-lysine affixing reagent to the sample target area.2. Allow the Poly-L-lysine to dry.3. Change the tip of the micropipette to avoid contamination.4. Using the micropipette place 0.5 μl of Gold colloid to the sample target area.5. Allow the Gold colloid to dry.6. Record a minimum of six (6) spectra of different regions of the enhanced sample target area using the FORAM.7. Repeat this method for other samples.Appendix III – Routine Quality Assurance ChecksQuality Assurance checks are performed on the FORAM to:• Check that the instrument is operating correctly before use.• Demonstrate that the instrument’s performance is consistent over a period of time.• Check that the supplementary materials are still providing accurate results.The checks are designed to demonstrate:a) The wave number accuracy of the instrumentb) The reproducibility of the instrument.c) The reproducibility of the associated materials.The following information details performance standards that are achievable using the Foster & Freeman FORAM and which may be used to help develop local Quality Assurance protocols. Each laboratory must develop and document a system of checks appropriate to their local working practices and quality requirements.Calibration CheckFrequency: DAILYQ.A. Test: Polystyrene standard check.Acceptable Result: Successful calibration.Quality Assurance of Gold ColloidFrequency: EACH TIME OF USEQ.A. Test: Benzyl Alcohol and Methyl Violet check.Place a solution of Benzyl Alcohol and Methyl Violet on to a piece of white office paper. Carry out SERRS analysis of this solution using the method described in Appendix II.Acceptable Result: The spectrum of the SERRS treated sample should be greatly enhanced compared to the non-enhanced sample.Blue spectrum: Methyl Violet without SERRSRed spectrum: Methly Violet with SERRSAppendix IV – Technical Information on Laser Spot Sizes and Fields of ViewThe size of the laser spot is fixed, so the area of the sample from which the spectrum is obtained is determined by the magnification of the microscope objective used.The table below provides details for the field of view and laser spot size for each of the microscope objectives supplied with the FORAM.Field of View VSC Suite Laser Spot Size (μm)Objective Lens X (mm) Y (mm)x 5 1.6 1.2 20x 10 0.8 0.6 10x20 0.4 0.3 5Appendix V – Application NotesApplication Note 1: Blue gel pensApplication Note 2: TonersApplication Note 3: Printer inks