Leica INM100 Optical Microscope Operation Manual
Contents
1. Startup Before turning on the microscope make sure that there is no sample on the stage that will collide with the objective lenses Find the power switch on the lower right rear of the main body of the microscope and flip 1t on Figure 1 This powers up the standard mercury illumination lamp for normal viewing and also powers on the electronics that drive the microscope Stage Positioning The manual stage 1s positioned by turning the knobs under the right front side of the stage table The gear engagement lever arm should be pushed away from to the side of the positioning knobs to engage the knob gears with the stage The lever arm pushed the other way enables easy and rapid manual positioning of the stage just by pushing it to an approximation of the intended location The upper knob moves the stage in and out relative to the operator Y Axis The lower knob moves the stage to the left and right X Axis The knob positioning is very precise and fine allowing positioning of very small objects into the field of view With the sample in place on the stage drive the stage manually to position the sample under the active objective lens Make sure that there will be no collision between the objective lens and the sample never force anything Stage Drive Selection lever Free Position Stage Drive Selection lever Engaged Position Y Axis Stage Drive knob X Axis Stage Drive knob Figure 3 Stage drive engagement lever pos2. CAB12006 6 foot S Video Cable This instruction manual Unpack all items carefully Check each item against contents list above Inspect unit for shipping damage If there was shipping damage call Microlmage Video Systems Immediately Do NOT plug unit in to power if damaged Further destruction and or injury may result Precautions Ls Do not touch the surface of the image sensor 2 Only use the specified power supply Use of a power supply other than the specified power supply will damage the camera 3 Do not attempt to aim the camera at the sun Never attempt to aim the camera at the sun or other extremely bright objects that cause smear to appear irrespective of whether the camera is operating or not This can damage the CCD 4 Do not allow the camera to be subjected to strong impacts or shocks DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 24 of 28 EA A Connectors and Controls po OQTg ee an The D108A camera back panel contains two connectors two rotary controls and one set of four DIP switches An additional set of switches is located under a narrow panel on the top of the camera body and may be accessed by removing the two small Philips screws The VIDEO connector provides a composite video signal which may be routed to a monitor via the supplied BNC cable The circular 12 pin connector marked DC IN accepts the mating plug on the PP108A power supply
3. extra magnification 1s set by the upper thumbwheel in the contrast selection module as shown in Figure 7 There are 3 magnification lenses in this section of 1X 1 25X and 1 6X To calculate the total magnification multiply the objective lens magnification by the extra magnification factor and then by the eyepiece magnification For example if you were using the 100X objective and the 1 25X transfer lens for extra magnification and viewing your sample through the standard 10X eyepieces you would have 100x1 25x10 1250X total magnification In addition to this there is a transfer lens between the microscope and the camera Figure 11 This lens will continuously add magnification from 0 3X to 1 6X to the total microscope magnification as you rotate the body of the transfer lens tube However there comes a point that extra magnification just causes loss of detail in your image so be cautious about how you utilize this feature Figure 11 Variable Magnification transfer lens Image Capture Software This microscope has a rudimentary video image capture system that allows us to capture an image onto a flash drive memory stick The resolution conforms to the standard NTSC video format not really appropriate for high resolution microscopy If you need publishable images use the INM200 microscope and its more comprehensive software DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page
4. 14 of 28 Basic Image Capture Procedure The basic procedure to capture an image starting from a dormant computer screen 1s as follows e Activate the Windows system by wiggling the mouse and clicking on the Computer icon e Click on the FB Spectrum icon in the system tray to bring up the image capture software This icon looks like a clear box with a big stylized red S inside o A small window will appear with the microscope image displayed The program name in the upper line of the window will be FlashBus Spectrum FBG 640x480 e To capture an image follow the steps o Click Grab in the upper right end of the tool tray This saves the image and displays the static capture image This is the place to do any editing of the image if necessary However the editing tools are quite rudimentary Click File and find your memory stick under the My Computer option Locate your target file folder on your memory stick and open it Type in the name you want to give the file o Click Save e To return the screen to the Live mode click the Live icon in the upper right tool tray O 200 Conclusion This microscope 1s a high quality manual microscope for general purpose examination of your samples It has very high magnification capabilities and also has a Fluorescence imaging capability as well as the full standard microscope features The scope is complex enough that incompatible feature selecti
5. DEFAULT Gamma 0 2 SW3 ON SW4 ON eS ee eee Internal Controls There are eight DIP switches located under a cover plate on the top of the camera body The first four of these are not used for normal operation and should be set to the OFF position Switches 5 6 and 7 are used to set the shutter mode or speed The available settings are Manual 1 60 sec SW5 OFF SW6 OFF SW7 OFF Manual 1 1000 sec SW5 ON SW6 OFF SW7 OFF Automatic Shutter SW5 OFF SW6 ON SW OFF Smooth Auto Shutter SW5 OFF SW6 OFF SW7 ON The back panel gain control will function normally in either of the manual modes When using the automatic fast shutter the camera will override the gain adjustment In Smooth Auto Shutter mode DEFAULT the control will override the automatic setting Switch number 8 allows the camera to display a negative image when in the ON position The camera produces a normal image when this switch is in the OFF position DEFAULT DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 26 of 28 Technical Assistance E EE For technical assistance contact WORLD VIDEO SALES CO INC P O Box 331 Boyertown PA 19512 Attention Customer Service Phone 610 754 6800 Www mivs com support mivs com Returns Microlmage Video Systems is a division of World Video Sales Co Inc Please call for a RMA Number and shipping address on all repairs WORLD
6. This connector also includes an S Video or Y C signal which may be routed from the 4 pin mini DIN connector on the PP108A to an appropriate monitor using the supplied S Video cable This is the preferred video connection IE A External Controls E EA Rotary Controls The control in the lower left corner of the panel labeled GAIN may be used to adjust the level of the video image in either manual or automatic smooth shutter modes The control in the lower right corner of the panel labeled COLOR may be used to adjust the red blue color balance of the image to compensate for different color temperature lighting systems DIP Switches The first two DIP switches on the back of the camera are used to set the level of edge enhancement This ranges from soft to very harsh vertical edges useful in bringing out details or determining a transition point Move the switch levers up for ON or down for OFF The possible setting combinations are Soft Edge SW1 OFF SW2 OFF Normal Edge SW1 ON SW2 OFF DEFAULT Moderate Edge SW1 OFF SW2 ON Hard Edge SWi ON SW2 ON DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 25 of 28 The third and fourth DIP switches are used to set gamma correction Move the switch levers up for ON or down for OFF The possible setting combinations are Gamma 1 0 SW3 OFF SW4 OFF Gamma 0 6 SW3 ON SW4 OFF Gamma 0 45 SW3 OFF SW4 ON
7. VIDEO SALES CO INC 625 Hoffmansville Road Suite 3 Bechtelsville PA 19505 Attention insert RMA number here Phone 610 754 6800 Note You MUST have an RMA number for returns amp repairs DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 27 of 28 Horizontal Resolution Image Sensor Scanning System Scanning Frequency Chroma Frequency Sync System NTSC Output S Video YC Output Shutter System Auto Shutter Range Gain Negative Image White Balance S N Ratio Minimum Illumination Gamma Aperture Lens Mount Connectors NTSC S Video Power Out Power Power Requirements Operating Range Dimensions Specifications ee AN 480 lines 1 2 CCD 768 H x 494 V 2 1 interlace RS 1 0 15 734 Khz H 59 94 Hz V 3 579545 Mhz Internal 1 Vpp composite video Y 1 Vpp C 0 28 Vpp burst Auto Manual 1 60 1 10 000 sec Manual On Off Manual better than 50dB AGC Off 0 72 Lux F1 2 5600 deg K 4 positions 0 2 0 45 0 6 1 0 4 positions C Mount BNC Female 12 pin Hirose HR10A male 12 Pin connector 12VDC 10 170mA 10 50 deg C 90 RH max 2 x 2 3 x 2 8 Simm x 59mm x 78mm Weight 6 7 oz 190g DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 28 of 28
8. microscope General technical background information on how a microscope works is contained in a lengthy document produced by Florida State University and can be found on their web site at http micro magnet fsu edu primer Introduction The INM100 is a semi automatic high performance optical microscope designed for the Semiconductor Industry It 1s equipped with a nominal CCD camera and a computer system to capture the images There is a large screen LCD monitor in the table with which to view and capture the images and a camera output LCD screen mounted on the wall behind the large screen The microscope is specially configured with an Ultra Violet Fluorescent module that will enable it to produce pseudo self illuminated fluorescent images of certain organic films that may be found on samples such as photoresist etc The microscope is somewhat complicated with many manual knobs to turn some combinations of which will produce total darkness through the lenses Therefore this document should serve as a solace of understanding in the midst of confusion and perhaps darkness Microscope Description Labeled photos of both sides of the microscope are shown in Figures and 2 It operates basically like this you look through the eyepieces to see a magnified image of your sample placed under the objective lenses on a manual mechanical stage illuminated by a mercury lamp at the rear of the microscope stand This microscope utilizes reflected l
9. the microscope body as seen in Figure 1 Figure 6 Light input Filter Selection pushrods There are three filters in this bank The rear push rod with no plastic ID cap is empty The next push rod is labeled Diff diffuser filter This filter is merely a frosted glass lens that insures that the illumination field is uniform in brightness It will reduce the light intensity by something like 20 The next push rod controls the DLF or Daylight Filter that compensates for the blue shift of the Mercury arc lamp source spectrum Inserting this filter will make the image appear slightly redder than the color without any filter The front push rod inserts the Blue Filter labeled BG 20 that accentuates the borders of the spectrum according to the instruction manual It makes the image appear with a bluish hue Viewing There are many ways of viewing your sample with this microscope 1 Look at it with the naked eye to see approximately where your pattern of interest is in relation to the substrate so you can position it under the objectives in the vicinity of what you want to see 2 Bright field illumination normal magnified view 3 Dark Field illumination shallow angle illumination which highlights topology so that the edges of the pattern light up and the flat areas appear dark 4 Interference contrast illumination where the contrast is enhanced or diminished by splitting the input light into two coaxial beams and
10. users have samples about the thickness of a 4 inch water This setting results in a thin gap between the lens and the stage so if you have a thick sample please take extra care with the system by lowering the stage inserting your sample setting the objective lens to a low power away from your sample then moving the sample under the lens and carefully bringing it into focus If you are viewing multiple thick samples you might want to reset the upper bound of the stage travel to prevent inadvertent collisions Bright Field Imaging Bright field imaging is the normal illumination mode for general purpose viewing This mode lights up the entire field with bright light so you can see your sample as if from directly overhead The bright field mode 1s selected by rotating the lower thumbwheel of the contrast function selection turret until it is latched into the BF DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 9 of 28 detent See Figure 7 The brightness of this mode is generally set by the lamp control thumbwheel on the front face of the microscope below the stage Extra Magnification Selector Thumbwheel Contrast Function Selector Thumbwheel Figure 7 Bright Field BF selection position in the Contrast Function Selection thumbwheel Dark Field Imaging Dark Field imaging is a technique that employs shallow angle illumination to darken flat areas but highligh
11. you to select either the NTSC default or PAL video i standard NTSC is used in the Western Hemisphere Japan South Ko e rea The Philippines and Taiwan PAL is used almost everywhere else ALLAI Figure 4 Input Type allows you to select the type of video input RGB com posite S Video or YUV depending on the type of camera consult the The Video Setup camera s documentation panel controls the appearance of live video and the video input type and standard DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 21 of 28 t KUE T a fae gt tal uE y E A ni EnA i lis Figure 5 The Grab Setup panel controls the appearance of captured images and flash parameters Effects Clicking on Effects displays a menu that allows you to rotate the video image clockwise CW Rotation or counterclockwise CCW Rotation Grab Setup Clicking on Grab opens the Grab Setup panel see Figure 5 which contains the following settings for flash grabs and grabbed images Brightness changes the brightness of a captured frame Contrast changes the contrast of a captured frame Field Delay indicates the number of fields to wait between a flash and a capture This number is typically anywhere from 0 to 3 Line indicates the number of lines to wait between a flash and a cap ture This rarely needs to be changed from the defau
12. E ains 14 Basic mase Capture Proceduro earn an e eE E i NSE 15 SAA no A A E E 15 ADP A a E E E O o A 16 SNC aN e VAC ico 16 The Most Frequent Faults in Microscopy occccccccnnnocncnnnnnonononanocnnnnnnnnnonnnnnnncnnnnnnnnnnnos 16 ls A E E E 16 Fa OAN RP ee on UNO II E a eeencencecordio st sat desea gee dueaesetcseeeondauvaaes 16 Un sharp Patches in the Microscopic mage oooonccnnccccnnonnnocncnnnnnnnnononnnnnnnnnnononnnnos 17 PP o uneseeceioaeease 17 DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 2 of 28 Lack of Fine Details because useful Magnification has been Exceeded 17 AN AMA T A A 18 BETAS TTS US POC eor 18 User Mangal TE A 18 OW 1 OMS italia ett 18 DOS Ose eee eee ee eee o een ener tere ent AA 23 Reference Material for Video Camera cccccccecccccsseccceseccceseccceeseceeunececeeceseeeceauaecs 23 DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 3 of 28 Leica INM100 Optical Microscope Operation Manual Roger Robbins 5 17 2006 C MyDocuments CleanRoomGeneral Equipment OoticalMicroscopes INM 100 INM100Manual doc Purpose The UTD Clean Room has purchased a new high performance optical Microscope from Leica Inc Itis a manually operated microscope and is called the INM100 This document gives brief operating instructions on how to physically operate our particular
13. Leica INM100 Optical Microscope Operation Manual Roger Robbins May 17 2006 The University of Texas at Dallas Erik Jonsson School of Engineering DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 1 of 28 Leica INM100 Optical Microscope Operation Manual Roger Robbins 5 17 2006 C MyDocuments CleanRoomGeneral Equipment OoticalMicroscopes INM 100 INM 100Manual doc Table of Contents es e o o E A EEA AEEA 2 Leica INM100 Optical Microscope Operation Manual ooonnnncncccnnnnnocncnnnnnnnnononannnnnnnnnnnos 4 PUTOS oscars ana E at taste 4 sn ee oe OE S 4 Microscope DESCUPIO Mba a roca 4 General Microscope Operation ccccccccnonnocccnnnnnnonononnnnnnnnnnnononnnnnnnnnnnnnnnnnnnnnnnncnnnnnnnnnnos 6 eR NN E o II II 6 A doe dare one gaa ett coonyne oe ee ercagctonao yeaa mitten cboacteee 6 OOD ECL Lens CVC CIO y MOP cdi ansaaereia serenrcasadesene rend PA 6 JEEE DAE A PU O on oO E 7 Pi A PI E S ese s aveanseses eee 7 A e O A 8 NC yy MAN eee se ae sone e E O eats oia cavemen oo ance a easton asa aces 8 FOO PEA O a o O A EE 9 Ba EC TVA ibas 9 Dark FPicld Aias N Tesne e a a e TE 10 Condenser Aperture Contrast Control ccccccccncnnoonnncnnnnnnnonnnnnnnnnnnnnnnnnonaninenoss 10 Intert rence Contrast TMA S110 sida a isis 11 Special Microscope Operati Ol errata indistinta 12 Fluoresconco Mario srr e E T RR 12 Extra Mastin odas 13 Mass O ELLA e E A O est A
14. e before you click LTE Enable The advanced Configure panel LiveMode not used with FlashBus Spectrim controls series captures and long Camera Type change to 3 when using a long term exposure camera term exposures You must select 3 before choosing LTE Enable see the Application Notes chapter for camera type information DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 20 of 28 Setup The Setup menu contains three choices for configuring video and image capture adjustments Limit Window Clicking on Limit Window limits the size of the video to a maximum of 640x480 in NTSC or 760x570 in PAL Video Setup a eel ie Clicking on Video opens the Video Setup panel see Figure 4 which contains a u the following settings for live video in FBG Brightness changes the brightness of the live video Contrast changes the contrast of the live video Saturation changes the saturation of the live video This is valid only for composite and S Video input Hue changes the hue of the live video This is valid only for compos E ite and S Video input i Sharpness changes the sharpness of the live video This is valid only o for composite video input Auto Iris changes the iris voltage level for auto iris cameras only This can also be controlled on the Grab Setup panel Standard allows
15. e with the vertical thumbwheel located just under the green light switch See Figure 8 DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 10 of 28 Figure 8 Details of illumination aperture control Green light out indicates that aperture size is under automatic control according to objective lens selection If light switch is toggled on then manual control with the vertical thumbwheel 1s in effect Interference Contrast Imaging This is a technique of introducing color hues by light wave interference to increase the contrast in the viewed image Basically the illumination light is split into two paths one impinges on the sample and the other bypasses it and then the two beams are recombined producing an interference effect much like the Michelson Interferometer Principle The result is a coloration of the image as various wavelengths of light reach constructive and destructive interference because of topological steps in the sample which cause light path length differences Along with this coloration contrast an additional 3 dimension like shadow effect appears that helps to bring out features of the specimen in greater contrast This function is selected by rotating the contrast function selection thumbwheel Figure 7 until it seats in a detent where the DIC label Differential Interference Contrast is showing In addition the DIC splitting prism thumbwheel setting Fi
16. eater storage space Figure 2 Source changes the camera displayed on the video screen The cam sis CON ere pane controls the size and era inputs are labeled 0 15 with 1 being the default a field aspects of the FBG video screen DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 19 of 28 Advanced Configuration zi Choosing Advanced Configuration on the Tools menu opens another Configure wai panel see Figure 3 which contains the following options Serial address not used with FlashBus Spectrim Remote Grab allows you to use a remote trigger for a flash grab with a compatible cable Series Delay sets the delay between each capture in a series For ex ample setting this to 10 causes FBG to capture every ten seconds Un der the Series Delay setting are the following options Series Off disables the Series Delay default Timed Series enables the Series Delay ee 7 Manual Series allows you to capture an irregular frequency of a frames as part of a series LTE Max Frames captures the indicated number of frames in a single long term exposure See the Application Notes chapter for more in formation about long term exposure SyncMode required for certain LTE cameras such as the Sony DXC950 and 970 Figure 3 LTE Enable enables the long term exposure You must change a the Camera Typ
17. g distances The fine focus knob is the smaller diameter knob that turns inside of the larger diameter coarse portion of the knob This is the tricky part of fine focus If the fine focus knob section is pushed coaxially to the left one rotation equals 80 microns of z axis movement If it 1s pushed to the right one rotation equals 20 microns of stage z movement This extra fine positioning of the z axis of the stage enables better control of focus for high magnification conditions The black ring next to the body of the microscope at the focus knob set is the stage z height lock This prevents the stage from moving too high and causing collisions with the lenses as they are rotated This is used in cases where the samples are thick and the stage needs to be low in order to achieve the proper focus gap between the sample and the lens If someone then wants to look at a thin sample the stage will not drive high enough to achieve focus This will require the black ring about the focus knobs to be loosened and the focus knob rotated to achieve focus with a high magnification lens and then re locked This however puts the stage at risk from the next user who may have a thick sample If one fails to take care in adjusting the stage for proper focus the stage and sample may collide causing breakage of either the sample or the lens or both The standard focus lock height is that which achieves proper high magnification focus for a 4 inch wafer since most
18. gure 9 must correspond to the proper objective lens See Table 1 Once selected the fine adjustment knob Figure 9 allows optimization of contrast by slightly changing the interfering light beam path length DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 11 of 28 Interference Contrast prism selector thumbwheel Interference Contrast fine adjustment knob Figure 9 Wollaston prism selection thumbwheel for Interference Contrast This setting must correspond to the selected objective lens to produce proper contrast The fine adjustment knob optimizes contrast by adjusting the interfering beam path length Table 1 Interference contrast prism selection vs objective Lens Prism Position Purpose Prism D Use for HC PL FLUOTAR lenses Mag 5x 10x 20x 50x which we don t have but it works ok with the FLUOTAR lenses above Prism C Use for PL APO lenses Mag 100x Normal position for bright and dark field illumination Prism D1 Use for HC PL FLUOTAR lenses Mag 5x 10x 20x 50x This 2 has a higher degree of splitting optimized for PL FL L Series objectives Special Microscope Operation Fluorescence Imaging This microscope is fitted with a fluorescence imaging system to allow detection of organics on substrates that will fluoresce from excitation by the microscope illumination This technique works in the following manner Extremely bright light from a sho
19. he Contrast Function Selection thumbwheel Figure 7 to the FL position Currently we have the red filter cube installed If you need the green one notify appropriate staff and we can physically exchange filter cubes for your application There is another requirement for fluorescence illumination and that is an extremely bright source lamp This is provided as a separate lamp added to the rear of the microscope upper body It is controlled by an external power supply as shown in Figure 10 To power on the Fluorescence lamp just turn on the Power switch It will take a number of seconds to warm up and reach full intensity When the lamp is at full intensity switch from the standard lamp to the Fluorescence lamp by pulling the lever out This rotates a deflection mirror and switches lamp sources Lamp Switch Lever Standard to Fluorescence Figure 10 Fluorescence Illumination lamp is on the left power supply on right Note that the life of this bulb is only 200 hours and it costs 200 to replace Therefore you must turn it OFF when you are finished Extra Magnification Occasionally you might want to have a little extra magnification to show an up close image of a specific aspect of your sample This can be done via the variable magnification transfer lenses which adjust the microscope internal magnification The DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 13 of 28
20. ight only compared to the INM200 microscope which can use transmitted light also good for photomask viewing Focus 1s achieved by rotating the stage focus knob and moving the stage with the manual drive knobs at the lower right side of the stage The light intensity is controlled by the thumbwheel on the front face of the microscope below the stage The other controls manage the more subtle capabilities of the microscope and will be described individually below DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 4 of 28 CCD Camera Zoom transfer lens Vario TV adapter Variable Magnification Dial Contrast Function Selector Dial Filter Selection module Wafer Rotation Table since removed Microscope Power Switch Stage gear engagement lever E Mechanical Stage drive knobs b Figure 1 Right side of Microscope Location of small Video display on wall Standard Illumination Housing Fluorescence lamp housing Lamp Adjustment Viewing Port Tiltable compensating Eyepiece Assy Focus Knob with Sensitivity Selection Objective Lens Selection Buttons Aperture selection button Fluorescence Lamp power supply Figure 2 Left side of microscope Standard Illumination adjustment on front face below stage DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 5 of 28 General Microscope Operation
21. itions Right freewheeling Left Gears engaged Objective Lens Selection To avoid collisions between the sample and the lenses lower the stage using the focus knobs to clear the lenses by a large margin The objectives are mounted on a motorized rotating nose piece that can be rotated to select the objective lens giving an appropriate magnification The higher the magnification the closer the lens comes to the DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 6 of 28 sample The lenses and nose piece are designed so that when the nosepiece rotates to a new magnification the image is still in focus Therefore the operator must start with the lowest magnification widest separation between lens and sample and focus After the low power focus 1s accomplished the lenses may be rotated to higher power using the lens selector buttons just to the rear of the left focus knob Figure 3 These two red buttons electronically rotate the nosepiece to change objectives The top button rotates the nosepiece from low power to high power and the bottom button rotates the nosepiece in the opposite direction a 3 Z Stage Max Height Lock Coarse Focus 15 i 5 Fine Focus e O O Figure 4 Focus knob and Red objective change buttons WARNING NOTE One should never rotate the lenses from the lowest power to the highest power The reason for this is that the lower power
22. lens has a large depth of focus making it difficult for the operator to achieve exact focus but the higher power lens has much less depth of focus and requires a much closer sample lens separation If the operator has set an imprecise focus separation on low power the high power lens could rotate into the sample crushing it and perhaps damaging the lens or rotator as well Lighting Lamp Power Since our normal eyesight has great difficulty seeing anything in total darkness we need light to illuminate the expected sample The Mercury vapor lamp for standard viewing is located at the upper rear of the microscope body It will light up when we turn on the microscope Its intensity is controlled by the little horizontal thumbwheel control knob located on the front face of the lower microscope body under the stage Rotate it to a numerical value of about 5 to 8 for a nominally illuminated field DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 7 of 28 aa Figure 5 Mercury lamp power thumbwheel horizontal orientation Filter Selection The last control over brightness is the light input filter pushrods as shown in Figure 6 These rods rotate filters into the lamp input path thus controlling the amount of light that passes through as well as selecting the light wavelengths input to the microscope optics By the way these are the pushrods located on the upper right rear of
23. lt of 0 Auto Iris changes the iris voltage level for auto iris cameras only This can also be controlled on the Video Setup panel Align aindicates which field should be grabbed first Any default is usually sufficient although Even or Odd is better when using a flash grab try both Even and Odd to determine which one is correct for the situation Type indicates the type of camera and flash unit used consult the camera s documentation DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 22 of 28 Appendix C Video Camera Operation Manual For Staff Only Reference Material for Video Camera D1081A DSP Color Camera Operation Manual Included for staff debug operations DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 23 of 28 Features _ 5 o el 1 2 CCD with 380K pixels produces image with horizontal resolution up to 480 TV lines Manual gain control e Adjustable white balance e Selectable gamma correction Negative image option e Composite video and S Video YC output gt Includes all necessary cabling and power supply Microlmage Support Unpacking AMA The Microlmage D1081A camera system for NTSC composite and S Video consists of Microlmage D108A camera module PP108A or equivalent Wall Plug power supply CAB11006 6 foot BNC cable
24. nfigure panel see Figure 2 which contains the following options Scale Video resizes live video to fit within the FBG window If you do E not select this only a portion of the video appears in the window Keep Aspect maintains the videos proportion even if the window is 4 resized If you do not select this the video is squeezed horizontally or vertically to fit in the window T af Field Rep eliminates uneven flash illumination on images Live video consists of two rapidly alternating fields one that displays video on odd numbered lines and one that displays video on even numbered lines With a flash grab one field is often illuminated more than the other i When you select Field Rep the even field is copied over the odd field to 2 even out the flash illumination sip One Field resizes video to less than or equal to 240 lines This is small enough that only one field is required Offscreen transfers video to the system memory first and then to the tak l VGA display memory as X Center adjusts the horizontal position of the video in the view screen f E to cover any black lines on the right or left sides Ji Y Center adjusts the vertical position of the video in the view screen 2 to cover any black lines on the top or bottom g es JPEG Q changes the size of files that you save as JPEGs Increasing the value creates higher quality JPEGs but requires gr
25. ngaged locked in position e Is the binocular tube set for your interpupillary distance and are the focusing eye lenses of the eyepiece set correctly e Is the optical system clean Uneven Illumination This can be caused by various errors To begin with check to see whether the revolving nosepiece and aperture diaphragm are in the correct position If not switch the microscope off and back on again and check for changes Remove all filters in the optical path which could cause vignetting in the image Check to see if the beam splitter of the Ergotube is in the correct position locked in place Check the lamp alignment Flat Images Defective objectives either produce no images at all or the images are flat or move when focused through It is often the case that front lens is damaged although the spring mount offers a high degree of protection Such objectives must be returned to the factory or to your local agency Do it yourself repairs usually compound the defect Dirty front lenses however are far more frequently seen This should always be the first suspicion when the image lacks contrast Finger prints and dust should be removed with a soft DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 16 of 28 cloth Resistant dirt should be removed with DI water Occasionally the eye lenses of the eyepieces should be cleaned They are often covered with oils from eyelashes Un
26. ons may produce no image at all This manual is intended to give you a basic understanding of the many features of the system so that you can successfully obtain revealing images of your samples Because of this complexity however we would like you to be trained by the clean room staff on this microscope before actually using 1t With this training and this manual you should be able to utilize the full capability of this microscope to reveal the secrets of your samples DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 15 of 28 Appendix A Suggestions from the Manufacturer Servicing and Maintenance e From time to time clean the outer surfaces and the contamination shield with a soft lint free cloth and distilled water with Isopropyl Alcohol in a 1 1 ratio e Accessible optics should be cleaned with a dry cotton swab Q Tip or lint free cloth e If the instruments and their accessories are still not clean do not proceed with further measures Please call the regional office of Leica for further instructions The Most Frequent Faults in Microscopy Before you start using the microscope ask yourself the following questions e Is the illumination correctly set e Is the lamp centered amp aperture diaphragm correctly set for the selected objective e Be sure that no filters are in the optical light path which do not belong there e Is the revolving nosepiece correctly e
27. rt arc mercury lamp is directed to a filter cube in the body of the microscope This filter cube filters the incoming light and passes a fixed band of wavelengths through the objective to illuminate the sample The short wavelength high energy light then excites the molecules that will fluoresce which then relax and emit a longer wavelength lower energy fluorescent light This longer wavelength must be separated from the exciting wavelength to prevent contamination of the signal with illumination background light DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 12 of 28 This 1s done in the same filter cube by a dichroic mirror which passes the fluorescent light but reflects the incoming illumination light Because the filtering action of the dichroic mirror is incomplete there is an additional bandpass filter on the exit port of the filter cube that sharply selects the wavelength of the fluorescent material that is finally directed to the viewing lenses of the microscope Now after having read the above paragraph it becomes apparent that this is a very specific light selection technique that must be matched to the fluorescent material you intend to examine Fortunately most fluorescent materials emit in either the red or green region of the optical spectrum and we have filter cubes that match both of these wavelength regions The fluorescence mode is selected by rotating t
28. s a reference however all of this information is available from the help file on the computer File and Edit The FBG file and edit menus contain standard Windows options Following are the notable options on the File menu see Figure 1 Save Settings saves the current FBG configuration for the next time you open the application To return FBG to default values click Reset on each appropriate panel as described in the following sections and choose Save Settings again Save Image saves an image in BMP TGA JPEG or TIFF file for mat Load Image retrieves saved images for viewing on the FBG win dow Series File saves a series of images First enable a series capture as described in the next section Then choose Series File type a filename Figure 1 and the series capture begins Files are given the name you entered ap The File menu pended with numbers that increase by one as the series progresses See contains standard the Advanced Configuration section of the next section for more about Windows saving and file series printing options DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 18 of 28 Tools The Tools menu contains two important choices pertaining to the FBG video window remote video captures image series captures and more Configuration Choosing Configuration on the Tools menu opens the Co
29. sharp Patches in the Microscopic Image Un sharp patches which remain stationary when the sample is moved are caused by dust etc on lenses and other optical faces The precise location can be seen when the eyepiece condenser deflecting mirror lamp condenser filter etc are rotated or moved With some experience it 1s possible to determine where the dust is located by observing how the dust patches move or not with the moving elements Here too cleaning should be carried out with a piece of soft rag or a soft brush Unnatural Contrast The aperture diaphragm contributes to the resolution and contrast An incorrectly set aperture can result in a too flat or too contrasty image with correspondingly reduced resolution The correct aperture should therefore always be ensured NEVER ADJUST BRIGHTNESS WITH THE APERTURE DIAPHRAGM Lack of Fine Details because useful Magnification has been Exceeded Excessive secondary magnification e g using the magnification changer at highest magnification as standard setting may produce empty magnification The image can then lack fine details Taken from the Leica Microsystems Operation Manual Version 1 4 10 2003 DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 17 of 28 Appendix B FlashBus Spectrum User Manual Excerpts Software Definitions A portion of the FlashBus Spectrum software documentation has been included here a
30. t edges and projections in the image field It is especially effective in illuminating edge steps in the pattern or particles on the surface of your sample It is selected by rotating the contrast function selector thumbwheel to the detent that shows DF on the flat of the thumbwheel Figure 7 Usually this technique also requires that the illumination intensity be increased either opening source apertures or simply increasing the lamp intensity However this microscope has an automatic aperture control which will attempt to compensate for the dark field darkness by opening the aperture Condenser Aperture Contrast Control The condenser aperture has a subtle effect on the contrast of the image By changing the size of this aperture the illumination cone projected into the objective lens is changed This affects the brightness edge contrast and resolution of the image The effective operating range is from 60 to 90 open Opening the aperture too much produces glare in the image and some loss of resolution Closing the aperture too much causes darkness and loss of resolution Physical control of the aperture is normally automatic green light off whereby the aperture is adjusted to optimum preset sizes according to the objective lens selected The automation even adjusts for bright field and dark field illumination automatically If your sample requires a different setting just toggle the green light switch on and adjust the aperture siz
31. then slightly adjusting the phase of one causing an interference color to appear over the image thus increasing its contrast 5 Fluorescence illumination with short wavelength light black light causing an organic material in the substrate to fluoresce give off its own light by down converting the incident high energy waves to lower wavelength red emission and 6 Combinations of the above All of these methods except the naked eye will also produce an image on the LCD display on the wall and on the large screen image capture software via the CCD video camera in the tower above the microscope body DOCUMENT TITLE Leica INM100 Optical Microscope Operation Manual 5 17 2006 DOCUMENT NUMBER SU2006 LI 001 Page 8 of 28 In any case look through the eyepieces to see the sample while driving the stage around with the right hand to locate your area of interest and adjusting the focus as necessary with the left hand you don t have to do anything with your feet Focus Focusing the image involves moving the stage up and down until the image is sharp in focus The Z axis of the stage focus is driven by the dual knob on both sides of the microscope base Figure 4 The focus knob set has three functions 1 coarse focus 2 fine focus and 3 max stage height lock to protect samples and lenses from collisions The coarse focus is controlled by rotating the large diameter knob section This allows the stage height to be moved rapidly over lon
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