PerkinElmer Spotlight 200 Page 53 User Manual Download

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Preparing Samples . 53

Tools for Sample Preparation

This section lists the tools you need for preparing samples:

•

Tools provided with the Spotlight 200;

•

Tools in the microsampling toolkit;

•

Materials to have available;

•

Specialized accessories you may want to purchase.

Tools Provided with the Spotlight 200

The following items for use in sampling are provided in the Sampling Accessories Kit that is

supplied:

Item

Use

Holder for 13 mm disks

Supporting 13 mm disks on the sample stage

Slides, glass (box)

Supporting samples for sample preparation

Rotating 13 mm disk holder

Supporting samples and allows rotation of the sample

Support for large samples

Clips on to the sample stage; supporting bulky samples so

that the stage clip does not interfere with them

Gold mirror assembly

Reflection measurements

KBr windows (2)

Supporting samples

Summary of Contents for Spotlight 200

Page 1: ...Spotlight 200 User s Guide MOLECULAR SPECTROSCOPY...

Page 2: ...s contained herein for incidental consequential damages in connection with furnishing performance or use of this material Copyright Information This document contains proprietary information that is p...

Page 3: ...ller 27 Overview of the Spotlight 200 29 A Guided Tour of the Spotlight 200 30 Connections 31 Operation 33 The Optical System 34 Visible Light Optics 34 Infrared Optics 37 Spotlight System Requirement...

Page 4: ...6 Removing the ATR Crystal Holder from the Microscope 87 Fitting a Crystal Assembly to the ATR Objective 88 Manual ATR Objective 88 Automated ATR Objective 88 Cleaning the ATR Objective Crystal 90 Aut...

Page 5: ...ide 5 Connecting the Microscope to the Spectrometer 122 Connections to the Stage Controller 123 Appendices 127 Appendix 1 Decontamination and Cleaning 128 Appendix 2 WEEE Instructions for PerkinElmer...

Page 6: ...6 Spotlight 200 User s Guide...

Page 7: ...Introduction...

Page 8: ...s Guide contain information on using and maintaining your Spotlight 200 Getting Ready to Use the Spotlight 200 gives you information on how to set up your Spotlight 200 at the beginning of the day s w...

Page 9: ...s two keys at the same time for example ALT F All eight digit numbers are PerkinElmer part numbers unless stated otherwise Spectrometer refers to the Frontier IR System Spectrum 100 Series Spectrum 40...

Page 10: ...n los recuadros como ste se proporciona informaci n sobre este tipo de circunstancias Caution Attention Nous utilisons le terme CAUTION ATTENTION pour signaler les situations susceptibles de provoquer...

Page 11: ...los recuadros como ste se proporciona informaci n sobre este tipo de circunstancias Warning Danger Nous utilisons la formule WARNING DANGER pour avertir des situations pouvant occasionner des dommages...

Page 12: ...l for the Spectrum Two spectrometer Spectrum Software Spectrum software has on screen Help which you can access by choosing the Contents command from the Help menu by pressing the F1 key or by clickin...

Page 13: ...Warnings and Safety Information...

Page 14: ...ide The Spotlight 200 The Spotlight 200 consists of a microscope spectrometer PC stage controller and joystick Figure 1 Spotlight 200 microscope and Frontier IR System Figure 2 Spotlight 200 microscop...

Page 15: ...body of the microscope or spectrometer switch off the power and call a PerkinElmer Service Engineer If your Spotlight 200 is fitted with an MCT mercury cadmium telluride detector DO wear safety glass...

Page 16: ...elative humidity 80 maximum non condensing If possible avoid any adjustment maintenance and repair of the opened operating instrument If any adjustment maintenance and repair of the opened instrument...

Page 17: ...ser serviceable parts in the microscope or the stage controller Capacitors inside the microscope and stage controller may still be charged even if the microscope or stage controller has been disconnec...

Page 18: ...cooling the system should not be sited near to room heating equipment for example central heating radiators There should be a minimum gap of at least 15 cm 6 inches from the top and side surfaces of...

Page 19: ...nstrument it means Caution risk of danger Refer to the manual to find out the nature of the potential hazard and any actions which have to be taken Microscope Safety Labels The following safety labels...

Page 20: ...or versions only NOTE See Cooling the MCT Detector on page 43 for information on how to fill the Dewar Stage Controller Safety Labels Figure 5 Rear of Stage Controller MAINS INPUT 100 240 V AC 60 VA 5...

Page 21: ...Information 21 Warning Signs on the Microscope Caution risk of electric shock Caution risk of danger Refer to accompanying documents to find out the nature of the potential hazard and any actions whi...

Page 22: ...Safety WARNING When you are using a motorized stage do not place your fingers between the moving and fixed parts of the stage The motors driving the stage from side to side front to back or up and do...

Page 23: ...ents Take care not to injure yourself or others or to drop the instruments The microscope weighs approximately 35 kg including the motorized stage Lift the microscope only by the base Do not attempt t...

Page 24: ...MC standard EN61326 EMC standard for electrical equipment for measurement control and laboratory use and EN55011 ISM class A rf emissions FCC rules and regulations This product is classified as a digi...

Page 25: ...such as large motors connected If possible do not use photocopiers discharge lamps radio transmitters and other equipment with large or frequent transient loads on the same supply circuit Microscope T...

Page 26: ...nt that is relatively dust free Make sure that the bench top is free from vibrations or mechanical shocks and is flat and level Do not place the Spotlight 200 near to room heating equipment for exampl...

Page 27: ...ge Controller Power supply 100 240 V 10 50 60 Hz 10 Primary fuse 1 6 A T time lag 250 V The control box is not designed for use with all three motors simultaneously over extended periods Maximum speed...

Page 28: ...28 Spotlight 200 User s Guide...

Page 29: ...Overview of the Spotlight 200...

Page 30: ...re enables you to find points on your sample and focus the microscope Two sizes of stage are available Sample sizes up to 75 50 mm part number L1860160 Sample sizes up to 160 69 mm part number L186016...

Page 31: ...output connections on the microscope and stage controller Figure 6 Connections to Stage Controller Figure 7 Electrical Connections top rear of imager Connections to stage controller SP1 connects to sp...

Page 32: ...way D type Stage Z Connects to Stage controller Z motor Connects to imager Drive Signals for the Z motor Connects to stage back connector Drive signals for Y motor stage Connects to stage front conne...

Page 33: ...les Transmittance Single areas of thin solids fibers and films The optimal aperture range depends upon the detector type as follows MCT detector Between 10 m and 100 m DTGS detector Between 50 m and 2...

Page 34: ...ttance and reflectance operation Dichroic mirrors form part of the optical system The dichroic mirrors used in the microscope reflect infrared and transmit visible light Visible Light Optics When you...

Page 35: ...es it at the sample position The upper cassegrain collects light from the sample and sends it upward through the aperture and the upper dichroic mirror Figure 10 Path of the Visible Beam for Viewing a...

Page 36: ...the aperture The lower transmission illuminator is also active in reflectance mode giving simultaneous illumination from above and below Figure 11 Path of the Visible Beam for Viewing a Sample in Refl...

Page 37: ...hen viewing a sample in transmittance as follows Figure 12 Instead of receiving light from the illuminator light from the spectrometer is reflected off the toroid onto the lower dichroic mirror which...

Page 38: ...gh the other side of the cassegrain toward the remote aperture The detector cassegrain focuses the beam on to the detector Figure 13 Path of the Infrared Beam for Collecting IR Spectra in Reflectance...

Page 39: ...ugh protection at a folder and file level to ensure that users and groups of users cannot delete or amend data files while at the same time being able to create new data files A graphics card with an...

Page 40: ...40 Spotlight 200 User s Guide...

Page 41: ...Getting Ready to Use the Spotlight 200...

Page 42: ...spectrometer is switched on Switch the stage controller and microscope on Cool the detector MCT detector versions only Set up the microscope Focus the microscope We recommend that you use these proced...

Page 43: ...exposure by wearing heavy gloves and safety goggles whenever you work with it WARNING When liquid nitrogen warms to room temperature nitrogen gas vaporizes so rapidly that resulting pressures can sen...

Page 44: ...liquid nitrogen Stand back and let the funnel empty completely The liquid nitrogen bubbles rapidly as it drains into the dewar This first amount of liquid nitrogen vaporizes completely as it cools th...

Page 45: ...id nitrogen settles down and bubbling slows 8 When the nitrogen stops bubbling refit the detector cap The filled dewar cools the MCT to the correct operating temperature for several hours After that t...

Page 46: ...agram and accessory toolbar are updated to show that the microscope is connected 9 Click the microscope button on the accessory toolbar 10 Make sure that the lower cassegrain is fitted that the ATR ob...

Page 47: ...ter and this has been setup to work in the near infrared you may find that there is a red tint to the image when viewed in the visible range To remove this fit a 1 T attenuator to the external beam wi...

Page 48: ...h a Frontier or Spectrum 400 FT IR FT NIR dual range spectrometer you will be provided with an Attenuator kit part number L1160560 containing 1 T 4 T 6 T 14 T and 32 T attenuators The attenuators atta...

Page 49: ...50 accumulations taken at 8 cm 1 resolution using an MCT or InGaAs detector 20 accumulations for a DTGS detector can be collected reasonably quickly For mapping or imaging experiments the total time f...

Page 50: ...y throughput Either manually adjust the lower cassegrain for maximum energy using the up or down buttons of the Correction function or use the Maximize Energy facility 4 Once the maximum energy throug...

Page 51: ...Preparing Samples...

Page 52: ...ransmission spectra the sample should ideally be thin enough approximately 5 to 20 m to give good detail and undistorted absorption bands The area of the sample must be large enough to give an adequat...

Page 53: ...items for use in sampling are provided in the Sampling Accessories Kit that is supplied Item Use Holder for 13 mm disks Supporting 13 mm disks on the sample stage Slides glass box Supporting samples f...

Page 54: ...s Picking up extremely small objects Roller knife Cutting knife end and flattening roller end Steel probe Pulling samples apart separating fibers Forceps 4 inch Cd plated Picking up small objects Tung...

Page 55: ...302613 NaCl 2 mm thick N9302614 KBr 2 mm thick N9302615 Wide tipped forceps hooked 09908138 Wide tipped forceps flat 09908400 1 5 mm microdisk fits in 13 mm disk holder to support very small samples 0...

Page 56: ...addition to the items provided with the Spotlight 200 we recommend that you have the following available Tape with adhesive on both sides double sided tape for holding long or large samples on the sa...

Page 57: ...m 1 The major disadvantage of this material is that it is hygroscopic so that the windows fog easily BaF2 Barium fluoride is not hygroscopic Its transmittance cut off is 750 cm 1 It can break or crack...

Page 58: ...as glass or metal A flat black cap from a jar makes a good surface for rolling a light colored sample If you roll the sample on a small flat piece of metal you can view it and collect spectra in refle...

Page 59: ...ecause the area of contact is small Rolling a hard sample with the side of a sewing needle held in a pin vise presses it into a flake Slicing Samples from Solids Cutting a wedge of sample Cutting a we...

Page 60: ...sed materials are paraffin wax This is the preferred medium for infrared spectroscopy It produces few spectral interferences and it can usually be easily removed from the sample with warm xylene pinen...

Page 61: ...ree screws and tighten them finger tight NOTE If the spectrum collected with the diamond anvil cell shows interference fringes place some KBr in the cell and collect a background spectrum through it F...

Page 62: ...er is left behind as ash 3 Score and break the pipette between the ash and the pyrolysate 4 Add a drop of solvent to the pyrolysate to wash it on to a salt plate If the amount of sample is small use a...

Page 63: ...simple map to help you positively identify the particles under the microscope Transferring with a tungsten needle When necessary sharpen the tungsten needle Nujol or fluorolube mulls Suspending fine...

Page 64: ...u can use a micropipette to apply liquid to the surface of a salt plate or to the edge of the junction between two salt plates In the latter case the liquid flows between the plates by capillary actio...

Page 65: ...Techniques for Collecting Spectra...

Page 66: ...to apply pressure across the windows The sample is held between the two windows The compression cell fits into the sample slide holder on the stage of the microscope Windows with thicknesses equal to...

Page 67: ...phase transitions this information is not available from thermal data only Polymers pharmaceuticals and liquid crystals are examples of materials where investigations of phase transition are important...

Page 68: ...g the Z control on the joystick CAUTION Once the lower cassegrain is removed lowering the stage when at the limit of its travel backwards will cause it to collide with the dovetail connector This will...

Page 69: ...he stage using the Z control on the joystick to its highest position 3 Click Zero on the Setup Microscope Advanced tab 4 Slide the cassegrain assembly back into the dovetail connector as far as it goe...

Page 70: ...rging system Some parts of the purging system are shown in Figure 22 Figure 22 Parts of the Purge System The purge system consists of The gas inlet connector on the metal plate at the rear of the micr...

Page 71: ...tlight 200 starting on page 41 3 Place the sample in position 4 Purge the microscope and spectrometer for 15 to 20 minutes at a rate of 10 l min 1 5 Collect the background spectrum and the spectrum of...

Page 72: ...f only those waves that have their plane of vibration in one particular direction The light that emerges is said to be polarized and is represented on the right side of Figure 23 Figure 23 Representat...

Page 73: ...change in polarization is dependent on the wavelength the color of the light emerging changes with the distance traveled through the sample and the amount of birefringence encountered Applications Dif...

Page 74: ...and their cylindrical shape can cause lensing effects For these reasons fibers are usually flattened in preparation for infrared microspectroscopy This flattening affects the birefringence of the str...

Page 75: ...he analyzer in with the wheel facing towards you It has two positions The first position allows the full beam to pass through When the analyzer is inserted fully the polarizing element is in the beam...

Page 76: ...parallel to or perpendicular to the bond direction For example stretching an acrylic fiber aligns the molecules with the general direction of the polymer chains parallel to the fiber axis and the nit...

Page 77: ...ope and the wheel facing outwards Figure 28 Infrared Polarizer Position 3 Push the analyzer into the slot until it stops 4 Turn the wheel to orient the polarizing element CAUTION The polarizer element...

Page 78: ...2 1 2 21 2 1 sin 2 n n d p p Where 1 n is the refractive index of the ATR crystal for germanium 4 0 is the wavelength of the radiation is the angle of incidence of the beam 21 n is the ratio of the re...

Page 79: ...at the sample position It is reflected once within the crystal then the totally internally reflected beam is collected by the rear portion of the upper cassegrain which focuses it on the remote apertu...

Page 80: ...e 5500 cm 1 to 600 cm 1 Ge Di 5500 cm 1 to 600 cm 1 Si 7800 cm 1 to 800 cm 1 Area of contact with sample Nominally 100 m diameter flat surface Single internal reflection from surface Maintenance The A...

Page 81: ...ly under the upper cassegrain Be careful not to change the alignment of the two adjusting levers 4 Fit the two adjustment thumb nuts Figure 30 The assembly is aligned if the adjusting levers have not...

Page 82: ...tlight 200 User s Guide Figure 31 The Automated ATR Crystal Holder and Cassegrain Jack plug socket plug not shown ATR crystal assembly Crystal holder Motor lowers and retracts the crystal Knurled thum...

Page 83: ...is directed through the microscope and not the spectrometer by using the Setup Instrument BeamPath tab 4 Place the slide holding the reference mirror supplied with the microscope on the sample stage...

Page 84: ...eference mirror by turning the bar clockwise and slowly lowering the toggle bar You must always support both ends of the toggle bar when lowering or raising the crystal OR Click the button to lower th...

Page 85: ...ng the knurled height adjustment nut left to right a quarter of a turn If there is a deep impression on the tape which does not disappear within a few seconds the crystal height should be raised sligh...

Page 86: ...2 Look at the image in the Camera View pane and focus the microscope on the surface of the mirror with the Z control on the joystick or the Auto Focus option There are usually dust particles or scrat...

Page 87: ...he Spectrum on screen Help for further information 3 Make sure that the ATR crystal is in the raised position 4 Screw the plastic protective cover in place over the crystal 5 For the automated ATR obj...

Page 88: ...older 5 Place the new crystal assembly into the ATR crystal holder and refit the height adjustment nut Take care not to damage the crystal 6 Refit the ATR crystal holder to the microscope as described...

Page 89: ...ine with the sprung loaded ball bearing in the wall of the holder Figure 37 You will feel the ball bearing slide into the alignment groove and lock the crystal assembly in position Figure 37 Crystal A...

Page 90: ...NOTE Do not use acetone or xylene to clean the crystal 1 Pour a small volume of a solvent into a shallow dish 2 Using the Z control on the joystick lower the sample stage to allow the dish to be place...

Page 91: ...s per year Every 4 5 months Medium User 15 000 cycles per year Every 8 months Light User 5 000 cycles per year Every 12 Months Materials Needed Methanol in a squirt bottle Small Foam Head Swab Recomme...

Page 92: ...embly crystal up on a clean work surface Remove the coned knurled collar to expose the crystal holder 5 Carefully grab the crystal holder and remove it from the ATR assembly Set the crystal holder asi...

Page 93: ...drain from the assembly 7 Spray methanol in hole above alignment bearing to flush out any free particles 8 Insert a methanol moistened swab into the larger gap between the bearing and the retracted pl...

Page 94: ...has evaporated it s time to reassemble the system Insert the crystal holder into the bore of the bearing being careful to line up the groove with the ball in the crystal holder 11 Gently press down o...

Page 95: ...ll set the lower limit of travel for the crystal 14 Reinstall the Auto ATR accessory in your Spotlight system and perform the crystal alignment and focusing procedure to maximize IR energy 15 Reset th...

Page 96: ...a battery When the battery power starts to decrease Spectrum displays a symbol in the Status bar at the bottom of the screen You should change the battery at this point If the symbol changes to the w...

Page 97: ...ridge battery 5 Replace the battery cover and tighten the screws firmly 6 Place the weighbridge on the stage and fix it in place with the four corner screws The PerkinElmer logo should be at the front...

Page 98: ...llecting a Sample Spectrum Once you have collected a background spectrum follow the procedure below to collect a spectrum of your sample CAUTION To prevent serious damage to the crystal do not move th...

Page 99: ...can and the spectrum is displayed as it is collected CAUTION Always retract the ATR crystal immediately after data collection has been completed in order to avoid accidental damage Scanning a Line or...

Page 100: ...has been completed in order to avoid accidental damage Automated ATR Objective Collecting a Sample Spectrum 1 Select the ATR sampling mode on the Setup Microscope Basic tab 2 Choose the Scan Settings...

Page 101: ...can change the size of each stage Z axis adjustment step using the options in the Setup Microscope Basic tab 12 When the spectrum is satisfactory click The sample spectrum is collected Scanning Marker...

Page 102: ...these will change the stage height during the analysis 5 Click Background spectra will be collected and the markers lines and maps will be scanned using the specified stage height The results will be...

Page 103: ...ercentage Sample Force reading in the Stage View tab and click Halt 8 Set the Auto Pressure slider to the required pressure 9 On the Setup Microscope Data Collection tab select the Determine Height Us...

Page 104: ...ed here assumes that you are familiar with using the ATR Imaging Accessory Further details of the installation and operation of the accessory are given in the ATR Imaging Accessory User s Guide L10500...

Page 105: ...registration mark clearly Figure 42 Image of ATR crystal registration mark NOTE From this point onward do not adjust the stage X and Y positions using the joystick 12 Select Microscope Set Stage Origi...

Page 106: ...ccount for the refractive index of the ATR crystal Refer to Adjusting the Aperture Sizes on page 107 19 Swing the crystal arm back over the anvil and clamp it in place on the pillars Ensure that the c...

Page 107: ...sses through the ATR crystal The ATR crystal is made of germanium which has a refractive index of 4 0 Adjusting the Aperture Sizes The crystal s refractive index causes the actual area scanned on the...

Page 108: ...nd maps to the image survey Make sure that the areas covered by the lines and maps are sufficent to allow for movement or distortion of the sample when it comes into contact with the crystal 3 Multipl...

Page 109: ...e of reflectance is seen in samples with matt surfaces such as paper and powders The broad collection angle of the microscope enables it to capture a large proportion of the diffusely reflected radiat...

Page 110: ...ion in one direction Figure 43 This is the type of reflection that occurs from a smooth polished surface Absorption information cannot be obtained directly from a specular reflectance spectrum of a di...

Page 111: ...ples Reflection Absorption Reflection absorption occurs when the incident radiation passes through a thin absorbent film that is on a reflective surface typically a metal and is then reflected back th...

Page 112: ...Maintenance...

Page 113: ...tage controller fuse Service This chapter contains a list of the available spare parts and optional equipment for the microscope If you need to replace a part use only PerkinElmer approved spare parts...

Page 114: ...d the excessive use of solvents Flowing solvents dissolve the cement on cemented optics dissolved cement can damage mirror surfaces Cleaning the Cover CAUTION Ensure the power is switched off and the...

Page 115: ...1 Switch off the microscope and disconnect it from the power supply 2 Insert a screwdriver into the slot at the side of the fuse drawer pull out and flip to one side over the mains inlet The fuse may...

Page 116: ...lowing manner Left Right joystick movement gives X movement of the stage Top Bottom joystick movement gives Y movement of the stage Twisting the joystick gives Z movement of the stage Left twist count...

Page 117: ...ns cord 2 Insert a screwdriver into the slot at the side of the fuse drawer and pull out and flip to one side over the mains inlet The fuse may now be removed 3 Fit the replacement fuse into the fuse...

Page 118: ...ng to keep it performing properly and to compensate for wear We recommend that the Spotlight 200 is cleaned examined and adjusted periodically by a PerkinElmer Service Engineer NOTE If you experience...

Page 119: ...y max 600 C 220 V 60Hz Korea L1860636 Cooler Accessory for use with L1860635 110 V L1860632 Cooler Accessory for use with L1860634 and L1860636 240 V L1860633 Rotatable Sample Disk Holder N1873039 Hol...

Page 120: ...ire Color 100 110 120 V Wire Color 220 230 240 V Ground Earth Green or Green Yellow Green Yellow Line Black Brown Neutral White Blue WARNING To ensure safe and satisfactory operation of the instrument...

Page 121: ...r 60 Hz and at voltages in the range 100 to 240 V without adjustment The stage controller will operate at all voltages in the range 100 240 V 50 or 60 Hz without adjustment Fit the molded connector of...

Page 122: ...ector marked on the rear of the spectrometer The connections from the pre amp in the microscope to the spectrometer are described below Pl 1 PCB D type Connector Line Power Requirement 1 11 Output Sig...

Page 123: ...OUT1 24 V max 2 52 A peak 2 OUT2 24 V max 2 52 A peak 3 OUT3 24 V max 2 52 A peak 4 OUT4 24 V max 2 52 A peak 5 X_END Y_END Z_END 5 V max Input signal 6 X_ZERO Y_ZERO Z_ZERO 5 V max Input signal 7 5...

Page 124: ...cribed below Pin Description Voltage Current 1 0VL 0 V 2 0VL 0 V 3 X_IN 5 V max Input signal 4 Y_IN 5 V max Input signal 5 Z_IN 5 V max Input signal 6 NC 7 NC 8 5 V 5 V 10 mA normal operation 2 4 A ma...

Page 125: ...Current 1 CTSOPT 5 V max 15 mA 2 RXD 5 V max 15 mA 3 TXD 5 V max 15 mA 4 DTR 5 V max 15 mA 5 0VL 0 V 6 CTS 5 V max 15 mA 7 RTS 5 V max 15 mA 8 CTS 5 V max 15 mA 9 ATR_stop 5 V max 15 mA 10 STEPSTAGE...

Page 126: ...126 Spotlight 200 User s Guide...

Page 127: ...Appendices...

Page 128: ...the Equipment Decontamination Procedure and complete the Certificate of Decontamination These documents are available on the PerkinElmer public website http las perkinelmer com OneSource decontaminati...

Page 129: ...effective means of protecting the customer s environment The requirements for waste collection reuse recycling and recovery programs are set by the regulatory authority in your location Contact your...

Page 130: ...tal 90 Auto ATR crystal 91 microscope cover 114 optical system 114 Cleaning the instrument 128 Compliance EC directive 24 EMC 24 FCC 24 Connections microscope 31 122 power 120 stage 31 stage controlle...

Page 131: ...ng 34 Sample preparation coatings 64 elastomers 61 fibers 63 fibrous solids 63 filled polymers 62 liquids 64 micro amounts 62 particles 62 polymers 60 techniques 58 Service 118 Setting up 46 Software...

Page 132: ...55 needle tungsten alloy 54 other useful 55 pin vise 54 probe steel 54 provided 53 roller knife 54 58 rotating disk holder 53 slides 53 support large samples 53 tungsten probe 64 tweezers 54 window s...

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