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1. Skip prt 0 ihe pes on RER Help B Dai ges The results will be displayed on the screen Note if a spectrum is saved with the resulting band fit the information will be saved with the spectrum for future reference Fit parameters lteration number number Options Search Allows initial start of iterations of the fitting Shape La d parameters to be procedure to be completed OE specified and or fixed before stopping SCH Skip pnt Increasing the SE free a number of skip data point SCHER bn P allows to check quickly the convergence of the algorithm and the quality of Opens a new window e i displaying the results of the initial value selection SS shi the band fit for Error displays the error Ee frt 0 Shapes e baselines and peaks value as distance between Error p position w full original data and curve sum Baseline width half max a amplitude or max intensity g Gaussian contribution s integrated area of all peaks and baselines This value is named usually as x2 Ref Doc UM HRSwHw En 45 72 Explore the future HORIBA HORIBAVJOB IN YVON 5 1 8 Band Integration lpr eg2 afble Full eo 29009 Format 3 Digits E A A A A mama oo IK ine ee be 5 1 9 Data size pcos This function reports the limits for each dimension of the active object in the following window and allows you to re define it if necessary The Size field rep
2. only activated data object is displayed Overlay seen all data objects are displayed on the same place Tile oan eessen data objects will be shown in the separated rectangles Projection trace of the activated data object will be shown in the separated rectangles Trace oe eessen single trace of the activated object will be shown Use the right scroll bar to change the trace v Dimension 1D 2D 3D v Use scroll scale Check this box to use the window bottom and right scroll bar v Always show scroll bar Check this box to always show the scroll bars when the visual zone is large then the data limits v One data per view If you check this box the next time data is displayed a new window will be created v Y scale X scale Uses the same scale for both axes useful for 2D image presentation Ref Doc UM HRSwHw En 94 72 Explore the future HORIBA HORIBAVJOBIN YVON Scale shows the scale properties for each of the window axes From value of low axis limit TO a value of high axis limit Freeze fixes axis limits The scale will be skipped when normalization function is applied Auto enables auto scaling when data object axis is changed Prop enables proper scale limits for each data object Log enables logarithm scale for the axis 5 3 2 Colors The Colors Dialog enables the image color properties to be modified Such properties are used when displaying images To acce
3. HORIBAVCOBIN YVON Correction Icon 35 Filtration Icon 36 EW Fourier filtering Icon 37 a i og l Mathematical treatments Icon 38 peak fitting Icon 39 KR Palette wWindOW Icon 40 Ka Spectral mapping procedure Icon 41 kab Profile Icon 42 RA Model procedure Ref Doc UM HRSwHw En 72 72 Explore the future HORIBA
4. HORIBAVJOBIN YVON LabRAM HR Software and Hardware User Guide HORIBAVJOB IN YVON SAFETY saaa A 3 1 1 GENERAL SAFETY INSTRUCTIONS ceszcisssnistavansqucuausayaaniniiwaiosusennsand lt iuuswendesanerasaqudinvaiawanesadsesanenndatedanidewanes 3 TE EEN 4 1 3 LASER RADIA HON SAFE EE 12 1 4 MAINTENANCE SAFETY INSTRUCTIONS ccccs0ssccceeceeseceeeeceeseceeeseeseeceeceuesceeeeeaeneceeeseaeaeceesesaneeeeeeaenees 14 1 5 SAFETY REQUIREMENTS FOR LASER SOURCE INSTALLATION 15 1 6 MECHNICAL SAFETY FEATURES ENEE REENEN EEN EEN 16 2 LABRAM HR DESCRIPTION conocida n a a a a E aA 72 AANT ODOC TON EE 72 22 PARTO DESCRIP ON EE 72 et WISI ION ANS oa E E E E E E E E R EEA 72 2 2 2 Configuration ofthe EE eet T2 220 Exo mal asos SOUC OS siose ee So E E E EE 72 2 2 4 Complete configuration for BX41 MICFOSCOPE cccccccescccessceesseeeseeseseeseseessseeseseessseeseseeseseeneneens 72 2 2 5 Basic configuration with BXFM Microscope 72 2 3 DESCRIPTION OF EQUIPMENT HARDWARE 72 20 1 erer ele ee 72 Ee E de EE J2 SS OPTICAL DESORPTION EE 72 2 4 1 Optical drawer CESCHONON ME T2 LALAO US OV OEE EA E E EEE A NOE E OAA 72 Te e ee 72 SEENEN 72 ZO OP TIGA SE CIR IGA TIONG EE 72 2 9 1 EENEG 72 Oe ISOC ISI and FOS ONO E 72 Ee 72 Ee 72 203 Reeg Ee ecan treet ee te Teer eee PEON Rec one ree one een teen te aoe ee ete Ee 2 6 POSITION OF THE DIFFERENT STEMS ENEE EEN 72 3 LABRAM HR MAINTENANCE AND USE 00 teeceeeseeennneeeeeeeneeeeeeeaaseeeseeea
5. 5 3 8 Scale bar The Scale Bar Properties dialog allows you to modify the display properties of the scale bar and its associated legend of the 2D presentations To access this dialog select Scale Bar item from the window context Scale Bar menu Options v Click on Line control to select the bar border style and color of the scale bar Y Click on Fill control to select the sacle bar z IR Cancel filling style and color Font 14 Times Color a v Click on Font control to select they the scale bar legend font properties v Click on Color control to select scale bar legend text color Sel Scale bar legend Ref Doc UM HRSwHw En 60 72 Explore the future HORIBA HORIBAJOBIN YVON 5 3 9 Image3D Style Internal IO External I Mode Overlay IMension 30 W Use scroll scale IT Always show scroll bars T One data per view IT Y scale scale To Freeze 391 605 EE r Ver bas 53 4 E r Intens H fi AEN Auto Prop WT r r The Image3D Properties Dialog enables the modification of the appearance of a 3D representation of the data object To open this dialog select the Image3D item from the window context menu v Type Select between polygon and gradation display mode v Gradation Select the axis which defines the direction of the color gradation v Click on the Color control to modify the polygon border style and color v Click on the
6. Test Banc Refresh time fo Binning factor i H Data mode Average Spike filter E T oee E 3 Single Scanning AJE Use detector Active D Double expo This is the more robust algorithm and works by comparing two or more accumulations within an acquisition If a random cosmic ray spike is observed the spike will be removed For this algorithm to work the accumulation number averaging must be set to greater than 2 Single pass This algorithm attempts to locate a spike in a single accumulation acquisition ie accumulation number 1 by analysing bands for sharpness width and intensity This algorithm needs to be used with care for very sharp Raman bands can easily be modified However the algorithm works very well when looking at broad features such as Raman bands of amorphous materials photoluminescence etc Note that both algorithms can be used for SIMPLE MULTIWINDOW single point mapping profiling etc Ref Doc UM HRSwHw En 34 72 Explore the future HORIBA HORIBAJOBIN YVON 4 3 2 Auto save via ACQUISITIONS in the menu bar The Auto Save window allows to modify the properties of the acquisition auto save option To access this window select Auto Save item in the Acquisition menu Auto save Save acquired data Format LabS pec ngs Folder e Memp EE Iw eat W Month Ww Dau Data Iw Hour jw Min Iw Count lo Help Cancel Save acquired data Select this box to
7. access it by selecting the Pointers item of the window context menu v Click on the Style control to choose one of the following pointer types Line single vertical line Pointer Cross vertical and horizontal lines Options Level vertical and horizontal lines Style The position of horizontal line corresponds to Selected the data intensity value ne E Cancel Double two vertical lines Deselected f Rect rectangle Keep E v Click on the Active control to choose the line style when the pointer is selected you can drag pointer with the mouse v Click on the Hidden control to choose the line style when the pointer is deactivated v Check Transparent box to draw a pointer in transparent style In such cases the image of the pointer is superposed with the window background Ref Doc UM HRSwHw En 99 72 Explore the future HORIBA HORIBAJOBIN YVON 5 3 7 Image The Image Properties Dialog enables to modify the appearance of the images data objects To access this dialog select Image item from the window context menu v Select Style mode to make or not a linear interpolation of image intensity along X and or Y axes Options m Help Style i o Hep Smoothed along column be Penal Unsmoothed _y Sa e Rye ey N Smoothed along diagonal Smoothed along line v Click Color to modify the image color This color will be used when you select overlay mode for the data display
8. Fill control to modify the internal polygon fill style Ref Doc UM HRSwHw En Image 3D Style Type s ge g ect Gradation Low Iw 2 i Lancel Color Fill 61 72 HORIBA Explore the future HORIBAJOBIN YVON Spim Pharma o Image 3D M Style Type E fe den Gradation je wv r z Lancel Color Fill Intensity omtisec Splm Pharma Image 3D m Style Type g E 8 den Help Gradation Jy x Yr 2 Cancel Color Intensity entsec Fill 62 72 HORIBA Ref Doc UM HRSwHw En Explore the future HORIBAJOBIN YVON 5 3 10 3Daxes The 3D Axes Dialog allows you to modify the appearance of the axes for 3D representation of data objects You access to this Axes dialog by selecting the Axes item from the window context menu v Options Options W Ticks Iw Values Label e MET Center PO Stile Frame KL Values 12 Times Ticks Labels 12 Times Help Cancel First select the axis you wish to customize Ticks Check this box to enable the drawing of ticks Values Check this box to enable the display of the number values for ticks Label Check this box to create the axis output label and edit the label field to customize the axis X lt gt Y Click on this button to switch between the X and Y axis Center Click on this button to re center the 3D object in the active window v
9. S SC Sg So Se Si S SCH Sex Sc S SCH o SCC eS SCENE S dy i Se ts ey e 3 SCH Gr CN SCH SCH SCH 2 SS S 3 Sg Se SC So e S dy Se S oe e ey 33 Sc ey Sc 2 Sc CN d i o CN CN 23 23 23 23 SCH lt 3 o Ce CN e ey dE 3 S Sex SCH Se See Ce a oS Gr e505 BIS 3 Sc SC d Sex CN o CH CH CN SCH SCH SCH ot e o SC 5 CN Sc So CN Go CN ep Sex 5c Se dy CN Gi e dE dE See Seege dE GEN Serres OOTO GEN eer erer EEN 10 72 Doc UM HRSwHw En Ref HORIBAVJOBIN YVON The label 12 is affixed on the 300x300 optional motorized XY stage to warn against potential risk of trapping fingers Ref Doc UM HRSwHw En 11 72 Explore the future HORIBA HORIBAJOBIN YVON 1 3 LASER RADIATION SAFETY A e On its basic configuration the LABRAM is a class 3B Laser Product A Class 1 accessory is available for complete housing of the microscope and the external laser source The LABRAM is supplied with an internal laser source eventually one or two external sources In case of addition of external source the end user must ensure that the laser source installation complies with all the legal safety requirements during operation maintenance and service For more information refer to 1 5safety requirements for le
10. YVON 4 THE LABSPEC SOFTWARE FOR DATA ACQUISITION 4 1 ACQUIRING A SPECTRUM The LabSpec software provides the user with a number of methods for acquiring a single spectrum Real Time Spectrum Adjustment This method is primarily designed for a fast display of the spectrum on screen corresponding to a single shot window The update is continuous providing a useful way of adjusting the focus position to maximise the Raman signal and quickly monitoring the stability of the spectrum Each spectrum displayed replaces the previously displayed spectrum there is no averaging or accumulation of the spectra and no extended coverage possible To do this you must use Spectrum Accumulation see below S Ensure the central spectrograph position has been correctly selected remember the position typed into the spectro window corresponds to the central position of the resulting spectrum e Choose the integration time required Click on the Real Time Spectrum Ie Adjustment icon S To stop the continuous readout click on the STOP icon on the top right hand side of the screen Ref Doc UM HRSwHw En 26 72 Explore the future HORIBA HORIBAVJOBIN YVON Spectrum Accumulation This method allows spectra to be acquired with multiple accumulations and averaging and or with coverage over extended regions Simple This method allows a single shot spectrum to be acquired with user defined integration tim
11. and accessories is identical for the smaller LabRAM 300 instrument The instrument can be considered in four parts 1 Lasers the HeNe 633nm laser is internal whilst other lasers are external mounted on an extended chassis at the back of the system 2 Microscope sampling is carried out through a standard optical microscope 3 Spectrometer dispersing the Raman signal into its constituent parts for detection by detector usually CCD but other formats can be used 4 Optics for coupling the lasers to the sample and carrying the Raman signal through to the spectrometer Spectrometer running OPTICS Lasers behind underneath top optics main unit 17 72 HORIBA Ref Doc UM HRSwHw En Explore the future HORIBAJOBIN YVON To provide the greatest flexibility and versatility the LabRAMs include a number of simple stems or push pull bars to allow fast switching between different functionalities Present on Present on LabRam HR Function LabRam Operates camera beamsplitter DOWN camera UP Raman LabRam 300 operates switching mirror for point mode IN and line scan OUT stem 2 LabRamHR operates switching mirror for vis NIR IN and UV vis OUT or point mode IN and line scan OUT Operates switching mirror for Stem 3 microscope IN and fibre entrance OUT labRam_ 300 operates grating turret in the LabRam HR this is motorised IN high resolution grating 1 OUT lo
12. selection In the Copy format column select the format Copy Paste Properties x Click on the OK button to validate the settings and close the window of the data exported to the clinhoard j Copy formiat e Paste format miaii ier i i unuunnuunnuunnuunnennnnnnnnnn nn Help Cancel C Data e Data e In the Paste format Text column select the format Click on the Cancel button to abort the changes and e Picture of the data imported from the clinhoard close the window e Data Native NextGen format Can be used to create the exact copy of the data object e Text Create the text spreadsheet of the active data object e Picture Create the screen Ref Doc UM HRSwHw En 64 72 Explore the future HORIBA HORIBAJOBIN YVON 5 4 3 Page Set U Page via FILE in the menu bar allows the print page to be configured including the spectrum itself information from the acquisition logo and comments v The Page window Page tools bar Components of the to print page ER Header page __ Associated tool Ay Page 04 Each icon allows to define the size and the position of Data object ES Associated tool each different predefined component type of the to print page Gaqraphic ob SC Associated tool a mage object d Associated tool ka Information parameters Associated tool ES v The page buttons load To l
13. the AP acquisition j The acquisition can be stopped at any time by clicking on the STOP Sron icon top right of screen Combining mode The Multi Window function allows to set the properties of the acquisition in multi window mode This mode enables to record data automatically over an extended range with a defined integration time and averaging The extended range is covered by taking a number of individual single shot windows and gluing these together To acces this Dialog select Multi Window item in the Acquisition menu From nasas nasenne Edit here the value of the first limit of the spectral window in the SE Den current spectral unit Select the following to activate the SEENEN DEER associated spectral window TO EE Edit here the value of the end limit of the spectral window in the Se EEN current spectral unit Select the following to activate the EE Ee associated spectral window TIME sess eier Edit the relative exposure time in of this window This value is RE EE used to compensate the differences of signal level Min overlap pix Define here the overlap between the individual windows in pixels ee A value of 50 gives generally good results SubPixel Selecting a value gt 1 allows to restore the fine shape features The EE EE method is based on the shifting of the spectrograph position on a EE EE distance less than the detector pixel size The result is to increase peewee EE the number of
14. the future HORIBA HORIBAJOBIN YVON Further specific details For time profiles the step size is in seconds and defines the time between the start of each acquisition Note that the overall acquisition time per data point must be less than the step size For depth Z profiles a negative number in the variation box implies moving the analysis point into the sample ie 20 gt 20 um will move 20 um below the start position and step its way up to 20 um above the start position The start position is always defined as 0 um Ref Doc UM HRSwHw En 33 72 Explore the future HORIBA HORIBAJOBIN YVON 4 3 OTHER ACQUISITION FUNCTIONS 4 3 1 Cosmic Ray Random spike Removal via ACQUISITIONS in the menu bar The CCD detectors such as those used on the LabRAMs are sensitive to other forms of radiation and in particular to random events known as cosmic rays These can interfere with acquisition of spectra by registering as very sharp and strong bands in the spectrum Since the occurrence of a cosmic ray is random it is extremely unlikely that a cosmic ray will occur in exactly the same part of the spectrum in two or more consecutive accumulations Hence it is possible to use a simple algorithm to detect random events in a spectrum as opposed to constant features such as a Raman peak In LabSpec there are two algorithms to choose from Options Exposition timelsec i Accumulation number nr Help Data name
15. you to modify the display properties of the spectrum Spectrum v Select one of the Style controls to display spectrum as a set of Lines Points or Bars syle iha L CW Help v Click Line control to modify the Line z Cancel color thickness and line type Options Point Sample Point Sample Intensity enker Lines Point Sample i te Kei A DA E S D l 1000 Raman Shift em t Ref Doc UM HRSwHw En 57 72 Explore the future HORIBA HORIBAJOBIN YVON 5 3 6 Spectra If the active object is a spectral image selecting Graph item of the window context menu opens the spectra Properties dialog which allows you to modify the display properties of the Splm object Splm Pharma SEs Spectra Options Line Help Intensity cntisec Fil C Cancel Lines No 1000 Raman Shift tech Click Line control to modify the color thickness and line type Click Fill control to modify the color and fill style of the filled zone Click Lines control to select the representation mode of all the spectra You can select to display All the spectra leave the software to display automatically the necessary spectra so it is readable or as shown on the picture above not to display all the spectra LAS 5 3 6 Pointers The Pointers dialog allows you to customize the appearance of the pointers You
16. 18069 2 Ast ACU Qa EY Microsoft Word Microsoft Power gt LabSpec m fe Myriam MOREA ERE EIE 10 24 Spectrum the spectrum at the current cursor Note the following descriptions are based upon analysis of a Raman mapped image but apply equally to analysis of a line temperature time or depth profile Ref Doc UM HRSwHw En 48 72 Explore the future HORIBA HORIBAJOBIN YVON 5 2 2 Analysing with Cursors The initial method for analysing a mapped image or profile is to use three cursors R G and B to define regions The image is generated by displaying the spectral intensity between these cursors For example in the above diagram the image can be seen to comprise three coloured regions which correspond to peak intensities in regions defined by the cursors The three cursors are simply selected from the three coloured cursor These three cursors appear when the window Splm is activated If one or more of the cursors aren t currently displayed on screen they can be returned by clicking on the Cursor normalisation on the toolbar ZS GF pe Px lt b ke J icon Using the cursors provides a fast method of getting useful information from the data However in order to distinguish different components this method does require that there are distinct non overlapping peaks which can analysed through the cursors 5 2 3 Analysing with Modelling With the modelling functionality
17. B version the laser beam remains accessible for a functional use below the microscope in the sample area The end user must ensure to operate in safe condtions with all the class 3B safety requirements Ref Doc UM HRSwHw En 12 72 Explore the future HORIBA HORIBAJOBIN YVON b The following precautions must be observed e Personnel must never look directly into the laser beam and should wear protective eye wear at all times if protective covers are removed while the laser is switched on e All personnel in the vicinity of the laser should also be ordered to wear protective eyewear if protective covers are removed while the laser is switched on Only qualified and trained personnel should be permitted to operate the laser Precautions must be taken to ensure that there are no reflecting objects in the path of the laser beam should protective covers be removed and safety devices deactivated Only beam stops made out of non flammable materials must be used Warning signs indicating the area in which the laser is enclosed should be clearly displayed e Local and national regulations governing the safe use of lasers should be adhered to at all times e Ensure that the laser is properly ventilated using a suitable exhaust Do not connect the exhaust to breathing air systems i e air conditioning or ventilation systems e Viewing laser beam with certain optical instruments eye loupes magnifiers binoculars or telescopes w
18. LES Introduction With suitable accessories on the LabRAM it is possible to acquire XY mapped images and line X or Y depth Z time and temperature profiles The principal of these measurements involves acquiring an array 2D or 1D of spectra with each spectrum acquired with a particular varied parameter for example position or time Variation Description Hardware requirement X and Y position map motorised XY stage X or Y position line profile motorised XY stage Z position depth or Z profile motorised Z stage piezo X Y and Z position volume motorised XY and Z stage piezo Time time profile no accessory required Temperature temperature profile heating cooling sample stage In all cases the spectral array 3D for volume 2D for map 1D for depth line time and temperature profiles is saved as a single file allowing fast and easy analysis of the data Ref Doc UM HRSwHw En 30 72 Explore the future HORIBA HORIBAJOBIN YVON Setting up a mapping profiling experiment With suitable accessories on the LabRAM it is possible to acquire XY mapped images and line X or Y depth Z time and temperature profiles 1 On the microscope select the objective to be used for the mapping experiment and select the same objective in the software LabSpec Yideo Test File Edit PMT Options Acquisition Data Setup Window Help 2 If an XY map XYZ volume or line profile is to be acquired
19. Labels Tick Check the box to enable the painting of mo a w a wm ticks La e R vi Grid Check the box to create a grid for tick lm levels E b B ntensity Values Check the box to enable the painting of Style number values associated to ticks Frame In Values 12 Times v Mam grid Labels E Times MM Sub grid Ticks 30 Look Labels Check the box to create an axis label and enable customization of the axis label In the Labels blank field enter the text you wish to be appeared for the ES corresponding axis l es ee Ref Doc UM HRSwHw En 96 72 Explore the future HORIBA HORIBAJOBIN YVON v Style Frame click on the control to modify the color and style of axis edge This parameter is used for a non 3D look Main Grid chck on the control to modify the color and style of the main grid Sub Grid click on the control to modify the color and style of the sub grid Values click on the control to modify the values font Labels click on the control to modify the labels font Ticks click on the control to modify the color and style of the ticks 3D Look check this box to enable a 3D outlook for the edge 5 3 4 GRAPH This function allows you to customize the appearance of 1D dimension displayed data objects Spectrum or Spectral Image 5 3 5 Spectrum If the active object is a spectrum selecting Graph item of the window context menu opens the spectrum Properties dialog which allows
20. Style Frame Click on the control to modify the color and style of axis edge Values Click on thes control to modify the font values Ticks Click on the control to modify the color and style of the ticks Labels Click on the control to modify the labels font Ref Doc UM HRSwHw En 63 72 Explore the future HORIBA HORIBAJOBIN YVON 5 4 OTHER FUNCTIONS 5 4 1 Multi via OPTIONS in the menu bar The Multi function allows whatever process is performed on the active spectrum to be performed on all open spectrum For example if a constant value is added to one spectrum with Multi selected all the open spectra will have that constant value added to them This can be a useful way to save a number of files with Multi selected a save dialog window will appear for each open spectrum in the order they are displayed in the Objects list 5 4 2 Copy Paste Via EDIT in the menu bar Copy CTRL X To copy the active data in clipboard You can select the data format with the Format command Paste CTRL V To insert the clipboard contents You can select the clipboard format with the Format command Format To select the Copy and Paste clipboard data formats This command permits to define the format of the data you export to the clipboard or and the one of the data you import from the clipboard To access this command select Format from the Edit commands menu in the Main menu bar v The Copy Paste data formats
21. abRAM must be protected by housing 5 The key must not be removed in ON position 6 11 Control that the emission indicator is always visible with the eyes wear Ref Doc UM HRSwHw En 15 72 Explore the future HORIBA HORIBAVJOBIN YVON 1 6 MECHNICAL SAFETY FEATURES Mechanical safety design provides protection against any hazards which could cause physical injury or burns Specific mechanical safety features are listed below H Exposed corners are smoothened e Air fans have grill guards with less than 4 mm access In the event of clogging of the fan grills these grills should be removed and cleaned replaced by Horiba Jobin Yvon personnel during maintenance interventions e No high temperature components are accessible to touch A Optional XY motorised stage case of the 300x300 XY stage It is highly recommended to take precautions when using this stage Indeed this motorised XY stage presents risks of trapping fingers The picture below shows the location of the dangerous parts of the stage They are 8 in total 2 in addition at the back of the stage directly opposite to the front part which are not visible on that picture A The highlighted parts can cause serious finger injury Ref Doc UM HRSwHw En 16 72 Explore the future HORIBA HORIBAJOBIN YVON 2 INTRODUCTION TO THE LABRAM HARDWARE The following diagram of a LabRAM HR system shows the typical layout The arrangement of stems
22. an button to apply the median smoothing The Median smoothing is a non linear data filter method Ref Doc UM HRSwHw En 42 72 Explore the future HORIBA HORIBAJOBIN YVON 5 1 6 Fourier transformation The Fourier Smoothing is based on the direct Fourier data transformation applying filter and apodization function and inverse the Fourier transformation The dialog window shows the real and imaginary Fourier functions and allows you then to select the smoothing property Fourier Fourier Data ok ai Help Cancel m Options Limit 50 Go Apod No sl Fiter No Limit Enter in this field the position of the cut off point in This parameter defines the smoothing factor 0 full smoothed 100 no smoothing Click on the GO button to apply the value or drag the pointer to select the Limit visually Apod Select here the type of the apodization function with apodization value reaching zero at a Limit point No apodization function Linear function Parabolic function Cosinus function Filter Select here the type of the filter function None No filter function Traffic Traffic function Ref Doc UM HRSwHw En 43 72 Explore the future HORIBA HORIBAVJOBIN YVON 14 5 1 7 Peaks and Bands operations iii Allows peaks within a spectrum to be quickly labelled and for particular bands to be fitted in order to accurately calculate band position width amplitude and in
23. and the external laser cover for the class 1 version Ref Doc UM HRSwHw En 5 72 Explore the future HORIBA HORIBAJOBIN YVON Label 5 DANGER The laser product class may increase in case of removing that cover and dangerous visible or invisible laser beam may become accessible if the covers are removed and the safety switch defeated FOR SERVICE PURPOSE ONLY Location On the microscope cover for the class 1 version and coupling chamber trapdoor Label 6 DANGER As microscope objectives are standard components there are not protected by a safety interlock Switch off the laser source before changing the objective Electrical and mechanical Hazard Label 7 Internal laser source label Location On the tube of the internal laser source Ref Doc UM HRSwHw En 6 72 Explore the future HORIBA HORIBAVJOBIN YVON Label 8 WARNING Screws has to be removed only for service Location On the top ot the main cover Label 9 DANGER Lasers are supplied by high voltage Always switch off and remove the power supply cord from the main before opening Take care that all capacitors are correctly discharged Location near the laser connector on the electronic rear panel and the external lasers Label 10 Grounding label Location On the rear panel of the main power supply Ref Doc UM HRSwHw En 7 72 Explore the future HORIBA HORIBAJOBIN YVON Label 11 Pr
24. asseeseoeaaseeesoaaseeessooanneees 72 3 1 OPTICAL ALIGNMENT AND MAINTENANCE OF THE LABRAM nn nsannnnsennnnnnsnnrrrnsnnrrrenennrrresrrrrrsenrrresennni 72 3 2 CHECKING OF THE SPECTROGRAPH LINEARITY sssennnsssennrressnnrrrrserrrrresnnriosnnrrrrsenrrrrrennrrresrnrrrerennn 72 3 2 1 Case of the LOA E 72 3 2 2 Case of EE 72 3 3 DIAGRAMM OF THE ZERO ORDER COMMAND sccccssssseececseasseeeecseasseeecssensesecseaaseesecseaaeeeecseaaeeeesees 72 ATE NOTCH FU Ete ee 72 3 5 CHOICE OF THE OBJECTIVES ccsccssessccsscsserscenccuserscenocssersencuseetecuscusersestoassersentoaueersensoseersensusersentoaues 72 OO eege 72 3 7 WROOBLE elen TEE 72 HORIBA Jobin Yvon Raman Division 231 Rue de Lille 59650 Villeneuve d Ascq France Tel 33 0 3 20 59 18 00 Fax 33 0 3 20 59 18 08 www jobinyvon com Ref Doc UM HRSwHw En 2 72 Explore the future HORIBA HORIBAJOBIN YVON 1 SAFETY 1 1 GENERAL SAFETY INSTRUCTIONS AN The LabRAM series equipment must not be operated without prior reading of this document The user manual contains important information on how to operate the LabRAM safely properly and most efficiently Observing these instructions helps to avoid danger to reduce repair costs and downtimes and to increase the reliability and life of the equipment Horiba Jobin Yvon equipment is perfectly safe as long as it has been properly installed and is operated according to the instructions which are given in this i
25. ata point s the spectrum of the selected data point s Click on the OK button to generate the map following the settings and close the mapping window SE Click on the Convert button to convert the current map into spectral image and close the mapping window Hel Click on the Help button to retrieve the information concerning 2 the parameters of this window Caneel Click on the Cancel button to close the Mapping window without validating the changes setting Ref Doc UM HRSwHw En 47 72 Explore the future HORIBA HORIBAVJOBIN YVON Before any analysis is made it is recommended that the data is saved In all cases ensure the window containing all the overlapped spectra Spectral Image or Spectral_ Profile is highlighted and save in the normal way A spectral image or profile is initially displayed with three active windows as below Splm containing all data of Map the resulting Raman the image profile image or profile MEIE LabSpec Map Blue File Edit PMT Options Acquisition Data Setup Window Help S S W SRP gt 0 GQ US Ge a ee Im oz Ok amp AS te tt SS Sl Map Blue ks Dele ra ke a Spim sample2 im Boe DA Intensity cnt Point Sample BS 4000 3000 S S n S S Ei 1000 Raman Shift cra Laser lt gt fi x fo 8 z ok KE of z fo E S For Help press F1 x 49 4059 37 1 i 1
26. bSpec 5 controls all the HORIBA Jobin Yvon Raman instruments enables different data acquisition modes eg Single spectrum multidimensional data set video image etc The data analysis routines include many commonly used Raman and FTIR tools such as baseline correction linear and non linearfilters peaks fitting Kramers Kronig transform etc The mapping techniques enables to generate and visualize map by using different spectral features band intensity peak position and so on The module architecture allows to customize software to support specific operation for example read foreign data file format implement data acquisition trigger etc The visual Basic script language of Microsoft is supported that yield possibility to create macro commands or include specific data analysis function The LabSpec 5 can be used also as Active X control in third part application The Main LabSpec screen can be divided up into a number of regions as shown below Menu Bar The main icons bar LabSpec Spectrum d03 File Edit Options Maintenance Acquisition Data Window Help Sar Si i SE nz Le ut R A tH Brae go ik amp te tt a Spectrum d03 SE x10 Graphical l Data object manipulation R H Za A D Da Oo RK w Ke X d e A Control panel Status bar l l 899 895 mei Ref Doc UM HRSwHw En 20 72 Explore the future HORIBA HORIBAJOBIN YVON 3 1 THE CONTROL PANEL The Control Panel l
27. data points defining a band ie 2 twice the SR EE number of data points 3 three times the number etc Note that EE this will increase the total acquisition time and it is suggested that RE EE sub pixel acquisition is only used over limited ranges 6000 sub 5000 4 S 5000 4 D ixel 3 4000 4 3000 4 2 30004 2 E 2000 20007 E T i T E T 510 515 520 525 Wavenumber cm 1 Ref Doc UM HRSwHw En 28 72 Explore the future HORIBA HORIBAJOBIN YVON Use multi window Select this box to activate the multi window acquisition mode Auto overlap Select this box to automatically define the overlapping size Merge data Select this box to create one data object for each active spectral window In the other case a data object is generated for each spectral range corresponding to the spectrograph position Adjust intensity Select this box to adapt the intensity to the same values on the overlapping parts This adaptation is based on the linear shift of the data Click on the OK button to validate the settings and to close the window Hep Click on the Help button to retrieve the information concerning the parameters of this window Cancel Click on the Cancel button to close the window without validating the change setting Ref Doc UM HRSwHw En 29 72 Explore the future HORIBA HORIBAVJOB IN YVON 4 2 ACQUIRING MAPPED IMAGES AND PROFI
28. e and averaging s Ensure the central spectrograph position has been correctly selected Choose the integration time required Choose the number of accumulations e Click on the Acquisition Options icon to select a cosmic ray random spike removal algorithm See page 34 Now click on the Spectrum Accumulation icon to start the acquisition s Multiwindow This method allows a spectrum to be acquired over an extended range with a defined integration time and averaging The extended range is covered by taking a number of individual single shot windows and gluing these together This procedure is fully automated through the software e Click on the Spectral Windows icon and choose USE MULTIWINDOW S In the SPECTRAL WINDOW PARAMETERS section type in the Start From and Stop To positions Edit the relative exposure time in of each window This value is used to compensate the differences of signal level Ref Doc UM HRSwHw En 27 12 Explore the future HORIBA HORIBAJOBIN YVON j j j iter l lit Spectro Select the integration time and be pie iee e a F number of accumulations from the ei Rief Ae E PE SE E Kr 3 Tem Test Z jo D tool bar at the bottom of the screen m T E P 2 x e Click on the Acquisition Options of the main menu to select a cosmic ray random spike removal algorithm See page 34 e Click on the Spectrum Accumulation icon to start
29. e are 6 neutral filters installed with the optical densities 0 3 0 6 1 2 3 or 4 no attenuation Po D0 3 Po 2 D0 6 Po 4 D1 Po 10 D2 Po 100 D3 Po 1000 D4 Po 10000 Confocal hole Reinitialise the hole sends the hole to a known reference point SE and then back to the position m displayed in the box I Open the confocal hole to its iah maximum value Close the confocal hole Type in the value for the confocal hole and oress return Slit Reinitialise the slit sends the slit l to a known reference point and als then back to the position Cl he sl z ose the slit bes d vi egener el m Open the slit to its maximum value slit and press return Spectro Reinitialise the gratings position move to the Zero order position L and back to the position displayed Send the spectrometer m Spectra z in the box to Zero order H b 8 s A Move the spectrograph grating to Wee Ne eieiei ate EEE fom the highest value of position retiirn Ref Doc UM HRSwHw En 22 72 Explore the future HORIBA HORIBAJOBIN YVON Select grating Select microscope objective ke Test lt File label prefix a file E name to each accumulated spectrum Exposure times nter here the exposure time value in seconds to be used for the spectrum and image RTD acquisition mode CH 5 lt lt 4 Enter here the exposure time value to be used fo
30. ectly by external optics or mirror In laser head and power supply there are high voltages which remain dangerous even if the device is disconnected from the main supply The following safety labels are affixed to the system and or to the optionel parts following different options Labels 1 to 7 indicate a hazardous situation where there is a risk of serious injury to be caused by laser radiation Label 1 The LABRAM operates with laser sources emitting visible or invisible continuous laser radiation typically bellow 300 mW The class of the laser product is 3B or 1 with the microscope and external laser sources housings DANGER AVOID EXPOSURE OF THE LASER BEAM TO THE EYES Or with the microscope housing or with external laser and microscope housing Ref Doc UM HRSwHw En 4 72 Explore the future HORIBA HORIBAJOBIN YVON Label 2 DANGER During operation the beam laser is emitted in the LABRAM Location On the main cover of the spectrometer and the microscope Label 3 A laser beam is emitted through that aperture DANGER Avoid exposure to the beam Location Above the microscope objective Label 4 The Labram has a protective cover which is fixed by screws This cover may not be opened or removed DANGER The laser product class may increase in case of removing that cover and dangerous visible or invisible laser beam may become accessible Location On the main cover of the spectrometer
31. ighten any screwed connections that have been loosened during maintenance and repair Any safety devices removed for set up maintenance and repair purposes must be refitted and checked immediately upon completion of the maintenance and repair work Spare parts must comply with the technical requirements specified by the manufacturer Spare parts from the original equipment manufacturer can be relied upon to do so Never modify the software of programmable control systems without informing and getting former consent from the manufacturer Ref Doc UM HRSwHw En 14 72 Explore the future HORIBA HORIBAJOBIN YVON A In case of changing of objective it is recommended to switch off the laser source before operating even if the access of the system have been secured by the means of enclosures equipped with approved security switches or firmly screwed shielding covers 1 5 SAFETY REQUIREMENTS FOR LASER SOURCE INSTALLATION All external laser source used by the end user have to follow the regulation CEI 60825 1 August 2001 To comply with legal regulations the following safety requirements need to be respected when installing the laser source po Class 3B X Emission indicator Certification and identification H Classwarninglabel X O e eres 2 training Jl _Eyeprotecton E JL Lasercontrolledarea X 13 Lasersafety officer X p14 Fire protection 1 The laser beam from the laser source to the L
32. it is possible now to distinguish spectral components that have a large number of overlapping bands and look at the distribution of any number of species with the cursors this is limited to just three one for each cursor The modelling algorithms are based upon correlation fitting of known reference spectra the models to the raw data These model spectra can in fact be obtained separately ie run spectra of known raw materials or taken from the mapping data itself Ref Doc UM HRSwHw En 49 72 Explore the future HORIBA HORIBAJOBIN YVON e With the map data open click on the Model icon on the toolbar BA If you wish to load in model spectra which you have already acquired separately open up these spectra e Activate the spectrum you wish to use as the first model component lf you wish to take this model component directly from the mapping data use the map cursor to find the spectrum you wish to use and then activate the raw data spectrum light blue in the Spectrum window e Click on Get in the Model window A new window named Score now contains from the models If the data is in overlay mode through FORMAT gt Mode in right click scroll down menu the image will display all the model intensities overlapped The Spectrum window now shows not only the raw spectrum at the current cursor position but also information about the correlation fit The coloured legends in the top right hand corner indicato
33. ithin a distance of 100 mm may pose an eye hazard and must therefore be avoided The sucking air fan located at the rear of the laser must not be blocked at any time Caution Use of controls or adjustments or performance of procedures other than those specified within this manual may result in hazardous radiation exposure The laser warning labels affixed to the system according to the safety regulations see warning chapter must not be removed The laser can only be switched on with the key switch This prevents inadvertent or unauthorised starting of the laser It cannot be operated with the key in the OFF position and the key can not be removed in ON position If mishandling of the instrument or of the safety devices results nevertheless in direct eye exposure to the laser beam the exposed operator should consult a doctor or a competent eye testing institution JD A security switch mounted on the sliding panel on top of the system which automatically activates the blocking of the laser beam when this panel is opened to intervene in the optical coupling drawer A screw is used in order to lock this sliding panel Therefore the use of a tool is required to open it When such an intervention is completed make sure to firmly screw it again Ref Doc UM HRSwHw En 13 72 Explore the future HORIBA HORIBAJOBIN YVON Laser Housing Laser safety devices must not be made inoperative except for defined checking procedu
34. laces the data intensities by sinus function The 1000 log abs x 1 converts intensity to logarithm scale The following rules are applied for the variables names the variables a b c correspond to the values of 1 2 data axis the x and y value are consider as intensity of data objects v Merge function Ant Click on the MERGE button to multiply data objects into a single Ref Doc UM HRSwHw En 38 72 Explore the future HORIBA HORIBAVJOBIN YVON 5 1 2 Baseline correction Allows removal of a background eg fluorescence from a spectrum Type of baseline rela oi approximation either R polynomial to be polynomial data points E used to lie on a polynomial Daai xl approximate curve or line segment lt baseline data points joined by SCH straight lines Clear Conver Attachment Yes data points forced to attach themselves to the spectrum No data points can be freely placed anywhere in window Build the baseline E Se l oeh ep SP Oe Erase the points of the baseline Once the baseline has been correctly approximated click on Sub to subtract the baseline from the spectrum Ref Doc UM HRSwHw En 39 72 Explore the future HORIBA HORIBAJOBIN YVON 5 1 3 Data correction Ti The Data correction function allows you to modify all the traces of a multidimensional data matrix Some of these operations require an additional spectrum as a second parameter
35. n the profile window Profile Profile S E S S Choose Hor or Ver control to select between horizontal and vertical profiles Click on the Corr button to replace the image profile This operation can be useful when you apply some operations to the profile for example smoothing and then you want to set data back to the image Ref Doc UM HRSwHw En 41 72 Explore the future HORIBA HORIBAVJOBIN YVON 5 1 5 Filtration d Ze This function allows linear and non linear smoothing and calculates the derivatives of first and second degrees The Linear Savitsky Golay smoothing and derivative computing are based on the convolution approach which performs a least squares fit of polynomial The larger is the Size value and the lower is the Degree value result in a higher smoothing effect Degree o Filtration Set in this field the degree of the f options polynomial The lower is the degree the more intense is the smoothing effect Size a Set in this field the number of the adjacent data points to be used for the calculation The larger is the Size value the more intense is the smoothing effect Smooth E Click on the Smooth button to perform the Savitsky Golay smoothing Der MEEO Click on the Der 1 button to calculate the first degree derivate ce eeben Click on the Der 2 button to calculate the second degree derivate ee pues Click on the Medi
36. name so that the Original map with complete data is not lost asa Search Open the Peaks amp Bands window S _ D Sizefpnt Moo Eat s Choose the band function you wish SE to use for the fitting procedure by Ge ve m clicking on the Shape scroll down Skipton F menu A baseline can also be Enr po added at this point SE e With a function selected now select the peak Label tool from the left a hand tool bar e On the Spim window mark which Options peaks you wish to fit Shape Search Level fi 4 d emmmmmmmmmmmmmmmmmmmmng S Click on Approx and then Fit to o start the fitting procedure Note this could take a few minutes Iteration 100 Se Skip pnt 4 to complete for a large map ma V Baseline CR Ref Doc UM HRSwHw En 91 72 Explore the future HORIBA HORIBAVJOB IN YVON Once the fit is complete click on Peaks e A new window will open containing details of the band parameters Fiting resulting from the fit o p Ss These include information on the SE baseline if present and all the do bands in the fit procedure The parameters are as follows p peak position cm a amplitude max intensity w full width half max em g gaussian contribution 1 max s integrated area of band The band parameters shown are for the spectrum at the current cursor position e Clicking on one of
37. nstruction manual The installation of the equipment is to be strictly carried out by personnel delegated by Horiba Jobin Yvon and should not be handled by the end user This manual must always be available whenever operating the equipment Any person carrying out measurements whether it is an engineer or an operator must be aware of the statements enclosed within this manual and apply its contents Tasks requiring familiarity with the manual include routine operation including setting up sample loading troubleshooting in the course of work Never make any modifications additions or conversions which might affect safety without the supplier approval This also applies to the installation and adjustment of safety devices In the event of safety relevant modifications or changes in the behaviour of the LabRAM during operation stop the equipment and namely the laser source immediately and report the malfunctioning to a competent authority person ex Horiba Jobin Yvon Service Ref Doc UM HRSwHw En 3 72 Explore the future HORIBA HORIBAVJOBIN YVON 1 2 WARNINGS CAUTION Performance of any procedures not specified by the manufacturer may result a hazardous radiation exposure This includes removal of covers with laser power ON There is no user maintenance or service internal to the system with power ON When covers or enclosures are removed for any reason extreme care must be taken to prevent the beam being viewed dir
38. oad an already recorded template Teg To save the current template into a file To validate the settings and to close the Page window Ee To close the Page window without validating the current Page settings v Using the Page tools icons Ref Doc UM HRSwHw En 65 72 Explore the future HORIBA HORIBAJOBIN YVON To insert a component in the to print page click on the corresponding tool Then move the mouse on the page so a cross appears Then move the cross at the wished location press on the left mouse button and move the mouse to define the size of the corresponding component v How to select a component of the to print page Move the mouse on the chosen component and click on the left mouse button Then a dotted rectangle is displayed around the selected component v Moving a component of the to print page Select the component Then when the following mouse cursor ck appears click on left mouse button and drag the mouse v Re sizing a component of the to print page First click on the object to select it Selection handles appear around the pe of selection J _ selected object Move the mouse pointer on one of the selection handles until to have the size pointer Ke vi Keep the mouse button pressed and move the selection handle and modify the size of the object v Modifying or creating the contents of a to print page component Except the data object the contents of all the othe
39. obtain a white light image of the sample and define the area line to be analysed Discrete points across a surface right mouse click to add and delete points Horizontal X ck Rectangular XY M gt H Sloped XY line Circular XY gt Vertical Y gt I i cs If a depth Z time or temperature profile is to be acquired simply ensure that the sample is correctly positioned beneath the microscope Ref Doc UM HRSwHw En 31 72 Explore the future HORIBA HORIBAJOBIN YVON 3 Click on the Spectral image properties icon in order to E define the mapping parameters Data Sizes From To Formule 0 8264 55 7051 p 7 4157 71 9101 i XY map set up by defined size or defined defined step will adjust accordingly and vice versa Area Volume mapping For area mapping X and Y dimensions need to be defined R E X Y Z For volume define the Z dimension as well Line profile X Z For horizontal line profile X gt dimension is defined aniv Ge _ Y Z For vertical line profile Y dimension is defined onlv For sloped line profile X and Y dimensions are active but size and step are defined only through the X dimension Time profile For time profile define time only Depth profile For depth profile define depth only Temperature profile For temperature profile define temperature only Ref Doc UM HRSwHw En 32 72 Explore
40. ocated on the bottom of the screen contains divisions which are directly related to the System configuration This panel will show only the devices which are installed on the System The description below details all of the available sub units Laser q mAlter SH m Hale pall pspectio E Soft IT 5 7 EI E e 2 m fie d i x o 8 we J a W ga d K giga Ob jy eo a d Krk um Kr um KE 1em Re Test O Aoo E Laser Laser Select the laser wavelength to be used from d fa pre set list Hehe M If your instrument is full automated Aramis instrument the set of the necessary optical pieces will be automatically set in place so the chosen excitation light reaches the sample In case this part is not automated on your instrument a warning message similar to the following on will be displayed LabSpec A Please select Laser 632 81 When everything is in place click then on the OK button Filter Click here to reset the filter wheel and to set Filter in place the filter displayed in the filter field Click here to open the list of the 6 neutral filters DS Ty installed Then select the one you wish to be in place Note The reset function of the filter wheel is useful when for any reason the selected filter is not well in place or does not match the selected one Ref Doc UM HRSwHw En 21 72 Explore the future HORIBA HORIBAVJOBIN YVON Ther
41. of the operation Correction i l 1000 2000 Ss Raman Shift em t Help From 506 372 To 1551 65 iv Limits H Noma E The Norma button normalizes all the traces to same area value 100 an eee The Zero button moves all the traces to the minimum intensity level Gel ee The Get button takes the activated spectrum and put it in Corrector frame This data object will be used as parameter WR The Del button removes the Corrector spectrum S NEE The Sub button subtracts the Corrector spectrum from all the traces i The Mul button multiplies all the traces by the Corrector spectrum oo ER The Corr button subtracts the Corrector spectrum from all the traces The intensity of the Corrector is multiplied to fit the intensity of the trace Limits Select the small blank box if you want that only a part of the trace is used for operation This parameter is used for Norma and Corr operation Limits From 506 372 To 1551 From and To Edit these fields to modify the limits of the selected region linked to the red cursors Ref Doc UM HRSwHw En 40 72 Explore the future HORIBA HORIBAJOBIN YVON 5 1 4 Profile Fa The Profile operation performs a visualization of the vertical or horizontal profile of an image To start a Profile operation e Load an image and activate image display window e Select cross line pointer by using window menu Pointer item e Click on the following icon to ope
42. oftware to use the Z motor for autofocussing e Piezo Force the software to use the piezo for autofocussing Z motor amp piezo The software uses the both devices for autofocusing A first approach is done with the Z motor The research of the final position is then refined by using the piezo Intensity Display the autofocus diode intensity Validate the settings and close the Autofocus window E Close the Autofocus window without validating the setting Ref Doc UM HRSwHw En 37 72 Explore the future HORIBA HORIBAJOBIN YVON 5 1 DATA ANALYSIS FUNCTIONS 5 1 1 Arithmetic 7 The Math Operation can be used to apply commonly used mathematical formulas to a data object Options Const Const Func 1 Func 2 v Addition of a constant value Enter the value in the Const field and click on the Const button to add the constant value to the data object v Multiplication by a constant value Enter the value in the Const field and click on the Const button to multiply the data object by the constant value v Application of a complex mathematical function Enter the mathematical function in the Funct or Func2 fields and click on the Funct or Func2 button to apply the corresponding function to the data object The mathematical formula can include basic mathematical function negating pow log exp sin cos asin acos atan abs sqrt step For example the sin a formula rep
43. on Icon 3 N Shape correction Icon 4 Zoom Icon 5 Intensity adjustment Icon 6 Shift Icon 7 le Manual addition Icon 8 Q Manual multiplication Icon 9 2 Manual labeling of peaks Icon 10 X Move peak maxima Fit peak width Icon 11 X Delete label of a peak Icon 12 H Integral calculation Icon 13 a build of a baseline Icon 14 Ref Doc UM HRSwHw En 69 72 Explore the future HORIBA HORIBAJOBIN YVON a delete the point of the baseline Icon 15 SS reduce the size of the graph background Icon 16 a delete the active object Icon 17 open a file Icon 18 al Se save a file Icon 19 Print page Icon 20 KR Information about object Icon 21 RE object normlalisatiion Icon 22 H Intensity normalization Icon 23 spi centering of the cursors Icon 24 IR Ref Doc UM HRSwHw En 70 72 Explore the future HORIBA HORIBAJOBIN YVON data sizes Icon 25 1 Informations on the active object Icon 26 Ie Spectrum adjustment Icon 27 BI CCD image adjustment Icon 28 d Spectrum recording Icon 29 RR Spectral images recording Icon 30 Val Automatic Multiwindow spectra recording Icon 31 ES Icon 32 CS Video image Icon 33 yt Baseline correction Icon 34 dh iA Ref Doc UM HRSwHw En 71 72 Explore the future HORIBA
44. orts the number of the spectral points for the corresponding axis From Ta Unit EES 253 337 2187 25 1cm EES 34272 Cnt Get 22 Click on the Get button to update the limits of the current active displayed object Scale o ESTE Use the Scale button to apply the limits to the active displayed object Extract __Evtract feed Extract the data included in the limits defined in the corresponding fields to build a new object Ref Doc UM HRSwHw En 46 72 Explore the future HORIBA HORIBAJOBIN YVON 5 2 ANALYSING MAPPED IMAGES AND PROFILES 5 2 1 Mappin tE The Mapping procedure generates a map by using the signal intensity in the selected spectral regions Up to 4 maps can be displayed when you load or acquire the spectral image the first three are the average signal intensity and the last is the signal ratio The selection of the spectral region is done with the Red Green or Blue pointer in the spectral image window Check Use box to Mapping CH You can edit From and To generate ge fields to adjust the limits of corresponding map ae the spectral regions Check Base box to a 7 subtract baseline d H HERZ S Green 1 24 04 1 355 36 Blue Iw 465 1 46 560 307 Check Green blue box to areen BUN j generate the map of the Spem M Conect Click on the Correct button corresponding ratio to modify data on the Check Spectrum box to see OF Convert Cancel selected d
45. otective earth label Location Inside the spectrometer and the main power supply Label 12 DANGER There is a mechanical hazard with the optional motorized table Always remove your fingers from the table when motors are in operation Location on the motorized table Ref Doc UM HRSwHw En 8 72 Explore the future HORIBA The labels 1 2 and 4 are affixed on the frame of the LabRAM near the CE stickers j Tea i H P L 8 mu Pees men EEN ier FEETES ssn EEEKA GAASS PSETES es Weg am Pe Manufacturer plate Label 4 keng a Weg hem GN or e TS Weg ot kend keng TS be The label 3 and 6 are affixed on the microscope aa wi IIIT IIIT HAREE i ARREA HARAR HARAR Label 3 Label 6 Ref Doc UM HRSwHw En The label 5 is affixed on the microscope enclosure and the slidding panel These parts are securised by the use of laser safety switches IT EZE eee eed seeete ened Label 2 S d Se SC d it e EE SE CN oe S 2 Se d S G Sc sd 2 SS Se d S oe dy SS S SC Ce 3 oe Sc S 2 GE SCH SCH Ce G Si 335 4 Gr CN Sc Ses cet dy Si Se Sex a SS Si ey CN Go d S Gi 5 SE EE SE ZS E ds Co SC CN e e ci 2 fe GE Sc tee e SCC S SE te S d ae ee CN 3 SC e Sc eS ty SC CN CN S
46. r objects could be modified by double clicking on the wished component Then a dedicated window appears with the tools to modify the corresponding component The options window for the header page the bottom of page and the Text object The parameters window for the array of experiment information parameters The Open window for the Image object Ref Doc UM HRSwHw En 66 72 Explore the future HORIBA HORIBAJOBIN YVON v Modifying the general page presentation or display Click anywhere on the to print page on the right button of the mouse to display the following menu Cut Cut To remove the selected component eee Format To define the parameters of the sheet of paper See Page Turn Setup zoom EI Turn To define the page orientation Portrait or Landscape Zoom To define the display of the to print page inside the Page window v The Parameters function On the printed document the selected parameters are laid out in an array The Parameters function permits to select the data parameters you wish to be printed and to define the style of the array To access this command double click with the left mouse button on the information parameters object when defining the layout of the to print document with the Page command v The Parameters window List of the non printed NSParameters tools to Print list List of the data parameters manage the list of the data data parameters parameters to orint selected
47. r the D gt spectrum and spectral image accumulation mode 2 Enter here the number of accumulations Motorized XY Z Table Position of the different axis Select the small blank box attached to of the motorized XY Z table E each table moving axis to have the Enter here also the position 0 E corresponding position value you want the table to be Z D refreshed in real time even when moved moving with the joystick Ref Doc UM HRSwHw En 23 72 Explore the future HORIBA HORIBAVJOBIN YVON 3 2 WHITE LIGHT ILLUMINATION IMAGE ACQUISITION The LabRAM is equipped with standard white light illumination of the sample by reflection and or transmission A colour camera linked to the software allows the sample to be visualised and the image captured on the computer and saved S Put the camera beam splitter into place DOWN camera UP Raman e Turn on the white light illumination e In the software click on the video icon to start the camera read out S To stop the continuous readout click on the STOP icon on the top right hand side of the screen Remember to turn off the white light illumination and take the camera beam splitter out UP before starting a Raman measurement To illuminate by transmission only remove the white light fibre optic from the top left port of the microscope Ref Doc UM HRSwHw En 25 72 Explore the future HORIBA HORIBAJOBIN
48. r the contribution of each component to the correlation fit to provide a match with the raw spectrum The coloured spectra displayed comprise Light blue raw data Red Green Blue etc models Black Fit Results of the fit 0 Raman Shift cm al H e Anew window will open up which contains the model spectrum S This process can now be repeated as desired Remember if model spectra are being taken from the map itself ensure the light blue spectrum in the Spectrum window is highlighted before clicking Get Ref Doc UM HRSwHw En 90 72 Explore the future HORIBA HORIBAJOBIN YVON 5 2 4 Displaying a map of band position width area etc The data contained within a mapped image or profile can be analysed by a band fitting routine and the results can then be plotted to give a map showing band position width area etc Such analysis can be of use when investigating changes in phase stress strain in materials and other such applications e With the map data open in the Spim window use the zoom tool to o select just the area which contains CH the bands which are to be fitted e Click on the Data sizes icon Sizes E select the limits of the spectral gt m CHE al Sch S SCH 1413 31 605 Lem region and then click on the Extract e l Crit button in order to remove all fno Je extraneous data and leave just the region of interest It is best that this extracted data is now saved with a different file
49. re laser power and alignment checking to be carried out by well trained engineers The LabRAM could be equipped with a main device that guarantees a class 1 product in terms of laser safety The CDRH enclosure mounted around the sampling area and embedding the microscope This enclosure contains laser safety interlocks with approved security switches which induce automated laser blocking when doors are open to access the sample holder 1 4 MAINTENANCE SAFETY INSTRUCTIONS Brief operating personnel before beginning special operations and maintenance work and appoint a person to supervise the activities The End User must ensure that the operators of the system are fully instructed in the safety procedures Ensure that the maintenance area is adequately secured If the LabRAM is completely shut down for maintenance and repair work it must be secured against inadvertent starting In any work concerning the operation conversion or adjustment of the LabRAM and its safety oriented devices or any work related to maintenance inspection and repair always observe the start up and shut down procedures set out in the manual and the information on troubleshooting and maintenance work Adhere to the procedures specified in the manual for routine checks and inspection troubleshooting and laser alignment checking For the execution of maintenance work tools and workshop equipment adapted to the task on hand are absolutely indispensable Always t
50. s options list Autofocus ia In the field you can adjust the shift value Ber Sen Then select if you wish to use or not the Autofocus and at which step of the acquisition process you wish to start the autofocus procedure EE Off The autofocus procedure is not used in the data acquisition EEN Process FOEI Before acquisition The autofocus procedure is applied before each data readout OOT Before each point The autofocus procedure is applied at each new measurement point positionning See Shift value he shift value allows to adjust the difference between the position where the laser reflection is at maximum level and the one where the spectroscopic signal is at maximum level This function allows you to define the properties of the Autofocus function To access to this window select Autofocus item in the Acquisition menu Ref Doc UM HRSwHw En 36 72 Explore the future HORIBA HORIBAVJOBIN YVON Autofocus Option From To Step Engine Intensity From Set the start position in um for the autofocus movement Re ao Set the stop position in um for the autofocus movement Step Set the step size in um for the autofocus movement Engine Allows you to select the device used for the autofocusing e Auto The software select automatically between the Z motor and the piezo following the size of the movement defined by the From and To parameters Z motor Force the s
51. save automatically the data when the acquisition process is finished Format Select here the file format The list of the different format supported is the same one as for Open Save functions Folder Edit here the path where to save the files Click on the d button to select the folder in the tree structure Year kee Select this box to add the year number to the folder name Month gt am Select this box to add the month number to the folder name Day nents Select this box to add the day number to the folder name File nana aes Edit here the file prefix name Hour aaa Select this box to add the hour to the file name Mm an Select this box to add the minute to the file name Count EL select this box to add the counter value to the file name The counter is modified each time the auto save function is called But you can edit the start value of the counter sequence Ref Doc UM HRSwHw En 35 72 Explore the future HORIBA HORIBAJOBIN YVON 4 3 3 Autofocus The Autofocus ensures an optimal focus is reached before an acquisition Click on the Acquisition of the menu bar Select Option item of this menu Check the box to activate the autofocus and click on the Autofocus button to set up the autofocus function This allows to adapt or not automatically the Z position to obtain a maximal signal level The process is based on the detection of the laser beam reflection Click on the down arrow icon to display the Autofocu
52. ser source installation m Laser sources used with the LABRAM constitutes a hazard to personnel during periods of operating maintenance and servicing Lasers are high intensity light sources producing visible or invisible light at specific wavelengths This concentrated energy in a narrow laser beam may cause damage to biological tissues especially to eyes To use lasers safely it is important to understand that lasers required final product safety classification is determined by essentially three elements the laser s power wavelength and housing Class 3B laser system Laser beams and reflected beams could be dangerous Diffused reflected beams in most cases do not present any problem Class 4 laser system Laser beam represents an acute hazard to the skin and eyes from direct and scattered radiation Fire hazard must be considered m The various parts of the LABRAM giving access to the laser beam have been secured by the means of enclosures and tubes from the laser source input to the microscope These tubes are firmly tightened and prevent any exposure from the operator to any laser radiation In the optional class 1 version an housing protect all the microscope and another housing encloses the external lasers m In case of addition of laser sources the beam path from the additional laser source to the LABRAM must be secured by the user with an adequate housing see 1 5 Safety requirements for laser source installation m In class 3
53. ss this function click on the right mouse button and select Colors menu items Colors Select here the active color frame W Red W Green Cancel jf Blue Neg Adjust Contrast and Brightness by moving the scale bar cursors In the Colors frame you can turn on off Red Green or Blue color To make image negative choose Neg check box In the Palette frame select one of the following styles rue color wsdecseadscounsesndennes All colors are displayed without modification Black and White False color Green oo eee eeececececeeeceeeeeees Yellow scale Ke OR Magenta scale EIS Cyan scale Blue senersrakenereakr rEranoN Magenta scale IT EE Cyan scale Red eae these dee ee Yellow scale Blue Lee eeeeeeceeeeececeeeeeees Green scale Ki D Green scale EISE Red scale 5 3 3 Axes Ref Doc UM HRSwHw En 55 72 Explore the future HORIBA HORIBAJOBIN YVON The Axes dialog allows you to customize the appearance of the graph axes Spim Pharma 600 s00 1000 1200 Gr I Intensity fentisec S P oy LG Lu T H aa Ei E F a a 1 l rT 1200 Ticks Values Raman Shift Lem BOTTOM Axis Intensity As shown on the picture above there are 4 axes The Axes Dialog allows you to define independently each component of each axis You access to the Axes Dialog by selecting the Axes item of the window context menu v Options Options Ticks Grd Values
54. tegrated area Peak Labelling To activate the peak labelling algorithm either click on the Search button or adjust the Level and Size pnt scroll bars These two scroll bars control parameters used within the searching algorithm for locating peaks in terms of the peak size and proximity to other peaks Adjusting these scroll bars will allow the labelling to be optimised Options Shape Level f1 Clear the labels from the spectrum Size prit Bo Fitting Wars Iteration 1 0 Check Baseline box to Skip pnt 0 Shapes fit baseline together with Error E the peaks nlk Baseline This allows the display to be controlled Note if a spectrum is saved with peak labels the peak label information will be saved with the spectrum for future reference Ref Doc UM HRSwHw En 44 72 Explore the future HORIBA HORIBAJOBIN YVON Band Fitting Options e Select or edit Peak Shape control to modify the peak curve shape e Select the function to be used and add a baseline if necessary SE ae o _ 11 Size prit An PE ONEI Iteration io Se Skip pri f0 O E Error Fra Close e Click on the peak Label icon and mark which peaks A are to be fitted M Options Search Shape Lett sl r SE em e Once all the peaks have been marked click on Level 11 Approx and then Fit geg 13 ot EH m Fitting Wars lheration 0 kou
55. the box noticed MAP will generate a new mapped image giving information about that parameter For example a map could be generated showing the exact peak position of a band according to the coloured scale for example dark regions gt low wavenumber position bright regions gt high wavenumber position Ref Doc UM HRSwHw En 92 72 Explore the future HORIBA HORIBAVJOBIN YVON 5 3 DATA DISPLAY FUNCTIONS 2 S S S E Hap Blue 20 P 30 D II J 40 e s Format so Norma Colors Y um Awe mage Scale Bar Pointer Ref Doc UM HRSwHw En Format Horna Scroll down menu Colors for spectra display Ames Graph Pointer Format Horma Scroll down menu for Colors image display Ayes Image Scale Bar Pointer 93 72 HORIBA Explore the future HORIBAVJOBIN YVON 5 3 1 FORMAT Style Internal H we p External Il e Mode Single MM Cancel Dimension 20 i Use scroll scale Always show scroll bars One data per view scale scale To Freeze Auto Prop Log Hor 20 0 w p a Ver 50 250 wo p E Intens 12 wo I E v Internal Click on this control to modify the color and style of the data field background v External Click on this control to modify the color and style of the window border Usually it is where the axes are painted v Mode Select one of the following data display modes Single
56. to be printed Click on the Style button Parameters ead to open the NSOptions window to define the Enter in the Title field the printed name of SC E E il design of the array the parameter Condition Date selected in the print D ee w click on the OK list Spec width P Spuet Ge d Cancel button to validate the Configuration Accumulation settings and to close Increment reen v the window Enter here the E s EE Click on the Cancel number of columns STE P DEEG of the printed data d to cl parameters array e ee EE the window Ref Doc UM HRSwHw En 67 72 Explore the future HORIBA HORIBAVJOBIN YVON gt Example of a printed data parameters arra y corresponding to the settings of the parameters screen copy as it appears on the printed document psure is the the exposition parameter Ee SE EE You will find below the data parameters arra Remark A column contains 2 fields the data parameter label and the data parameter value gt The Parameters tools EI Add the selected data parameter to the print list Remove the selected data parameter from the print list inthe print list move the selected parameter to the previous line Blin the print list move the selected parameter to the next line Ref Doc UM HRSwHw En 68 72 Explore the future HORIBA HORIBAJOBIN YVON Icon 1 K Standard cursor of work Icon 2 L Peak eliminati
57. w resolution grating 2 Steins LabRam HR operates switching mirror for two detectors IN side detector OUT top detector y always present o sometime present Ref Doc UM HRSwHw En 18 72 Explore the future HORIBA HORIBAJOBIN YVON In conjunction with the software the LabRAM control box provides an interface between the user and the instrument An explanation of the switches on the front panel is given below Laser shutter switch Internal diode UP Shutter i back Line scan open laser on a ignment facility sample UP ON UP ON DOWN laser DOWN OFF DOWN OFF blocked by shutter Detector UP shutter Main key control Shutter switch Camera power controlled by for internal UP UP video detector 1 633nm laser microscope camera on DOWN shutter off position DOWN fibre DOWN video controlled by shown entrance camera off detector 2 SHUTTER LASER OFF DIODE ON SHUTTER ANNER ON CAMERAON LASER ON SHUTTER 1 DRU LASER INTERLOCK i SHUTTER 2 Power indicators for laser and interlocks on when lit Power indicator on when lit For more information on the hardware including optical layout please see the LabRAM User Manual Ref Doc UM HRSwHw En 19 72 Explore the future HORIBA HORIBAVJOBIN YVON 3 INTRODUCTION TO THE LABSPEC SOFTWARE LabSpec 5 is a general data acquisition and data analysis software based on open NextGen module architecture La

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