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Care and Use of the BrightCare and Use of the BrightField MicroscopeField MicroscopeMalaria parasiteMycobacterium tubercul...
Importance of the MicroscopeImportance of the Microscope• Important for hematology, microbiology, TB, andmalaria testing• ...
Basic Components of the MicroscopeBasic Components of the Microscopeand their functions...and their functions...Power swit...
Care and Maintenance of the MicroscopeCare and Maintenance of the Microscope• Good preventive maintenance and care include...
Cleaning the MicroscopeCleaning the MicroscopeRoutine Cleaning Supplies:Routine Cleaning Supplies:• Commercial lens tissue...
Cleaning Oculars and ObjectivesCleaning Oculars and Objectives• Unplug the microscopeUnplug the microscope• Wash handsWash...
• Unplug microscope and allow bulb toUnplug microscope and allow bulb tocoolcool• Carefully place microscope on its sideCa...
• Plug in microscope and turn on illuminator. RotatePlug in microscope and turn on illuminator. Rotatenosepiece to lock 10...
• Close field diaphragm half way and focus smear at 10XClose field diaphragm half way and focus smear at 10X• Close field ...
Operation of the Microscope –Operation of the Microscope –Examining SmearsExamining Smears• Put smear on stage and center ...
Examining SmearsExamining Smears (continued)(continued)• Adjust sharpness of image by moving adjustmentAdjust sharpness of...
Microscope Problems – Troubleshooting 1Microscope Problems – Troubleshooting 1ProblemProblem: Black Field: Black FieldPoss...
Microscope Problems – Troubleshooting 2Microscope Problems – Troubleshooting 2ProblemProblem: Field only partially illumin...
ProblemProblem: Difficulty focusing with 10X: Difficulty focusing with 10XobjectiveobjectivePossible Causes:Possible Cause...
Microscope Problems – Troubleshooting 4Microscope Problems – Troubleshooting 4ProblemProblem: Difficulty focusing with 40X...
Microscope Problems – Troubleshooting 5Microscope Problems – Troubleshooting 5ProblemProblem: Blurry image at 10X or 40X: ...
CreditsCreditsThanks to STD/HIV Prevention Training Center ofNew Englandin Collaboration withState Laboratory Instituteand...
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Care of microscope

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Care of microscope

  1. 1. Care and Use of the BrightCare and Use of the BrightField MicroscopeField MicroscopeMalaria parasiteMycobacterium tuberculosis
  2. 2. Importance of the MicroscopeImportance of the Microscope• Important for hematology, microbiology, TB, andmalaria testing• Compound microscope used in bacteriology, biology,and medicine to examine minute objects such asbacteria, other unicellular organisms, and plant andanimal cells and tissue• Advances in fluorochrome stains and monoclonalantibody techniques caused growth in use offluorescence microscopy in both biomedical analysisand cell biology
  3. 3. Basic Components of the MicroscopeBasic Components of the Microscopeand their functions...and their functions...Power switchPower switchLight intensity controlLight intensity controlCondenserCondenserStageStageObjectivesObjectivesEyepiece lensEyepiece lensStage motionStage motioncontrol knobscontrol knobsCourse and fineCourse and fineadjustmentsadjustmentsknobsknobsField diaphragm ringField diaphragm ringAperture diaphragmAperture diaphragm
  4. 4. Care and Maintenance of the MicroscopeCare and Maintenance of the Microscope• Good preventive maintenance and care includes:Good preventive maintenance and care includes:• Regular cleaning of oculars and objectivesRegular cleaning of oculars and objectives• Avoid damaging oculars and other optics with eyeAvoid damaging oculars and other optics with eyemake-up or other debrismake-up or other debris• Careful handling to avoid abrupt motionsCareful handling to avoid abrupt motions• Protect from direct sunlight, high temperature,Protect from direct sunlight, high temperature,humidity, dust and vibrationhumidity, dust and vibration• Use appropriate materials to clean the lensesUse appropriate materials to clean the lenses• Cover when not in use with vinyl or plastic dust coverCover when not in use with vinyl or plastic dust cover
  5. 5. Cleaning the MicroscopeCleaning the MicroscopeRoutine Cleaning Supplies:Routine Cleaning Supplies:• Commercial lens tissue for opticsCommercial lens tissue for optics• Caution: DoCaution: Do notnot use paper towels oruse paper towels orother rough paper productsother rough paper products• Cotton swabs with wooden shaft (optics)Cotton swabs with wooden shaft (optics)• 70% isopropyl alcohol70% isopropyl alcohol• Dilute methanol is satisfactoryDilute methanol is satisfactory• Mild detergent and soft cloth for stageMild detergent and soft cloth for stageand base of microscopeand base of microscope
  6. 6. Cleaning Oculars and ObjectivesCleaning Oculars and Objectives• Unplug the microscopeUnplug the microscope• Wash handsWash hands• Remove dust from optical glass surfacesRemove dust from optical glass surfaces• Carefully remove eyepieces, objectives,Carefully remove eyepieces, objectives,condenser, and filters–one at a timecondenser, and filters–one at a time• Excessive rubbing can cause damage toExcessive rubbing can cause damage toiridescent coating on lensiridescent coating on lens• Clean and replace as completedClean and replace as completed• Do NotDo Not take eyepiece or objectives aparttake eyepiece or objectives apart
  7. 7. • Unplug microscope and allow bulb toUnplug microscope and allow bulb tocoolcool• Carefully place microscope on its sideCarefully place microscope on its side• Open bulb house; use tissue to removeOpen bulb house; use tissue to removebulbbulb• Use tissue (to avoid fingerprints) to pickUse tissue (to avoid fingerprints) to pickup new bulbup new bulb• Insert new bulb and close bulb houseInsert new bulb and close bulb houseReplacing Microscope BulbReplacing Microscope Bulb
  8. 8. • Plug in microscope and turn on illuminator. RotatePlug in microscope and turn on illuminator. Rotatenosepiece to lock 10X objective in placenosepiece to lock 10X objective in place• Place smear on stage and center it under the 10XPlace smear on stage and center it under the 10Xobjectiveobjective• Open the field diaphragm all the way and closeOpen the field diaphragm all the way and closecondenser diaphragm all the waycondenser diaphragm all the way• Move up (rack up) stage to its highest positionMove up (rack up) stage to its highest position• Adjust the oculars for interpupillary distance so thatAdjust the oculars for interpupillary distance so thatonly one circle of light is seenonly one circle of light is seen• Rack up condenser as high as possibleRack up condenser as high as possibleSetting the Koehler IlluminationSetting the Koehler Illumination
  9. 9. • Close field diaphragm half way and focus smear at 10XClose field diaphragm half way and focus smear at 10X• Close field diaphragm until diameter of illuminatedClose field diaphragm until diameter of illuminatedimage is smaller than the field of viewimage is smaller than the field of view• Lower condenser with positioning knob until you haveLower condenser with positioning knob until you havea sharp, focused image of the edges of the fielda sharp, focused image of the edges of the fielddiaphragmdiaphragm• Adjust condenser using centering screws so that theAdjust condenser using centering screws so that thecircle of light is centered in fieldcircle of light is centered in field• Open field diaphragm until illuminated image is justOpen field diaphragm until illuminated image is justlarger than the field of view. If more light is needed,larger than the field of view. If more light is needed,use the transformer.use the transformer.• Koehler illumination is now set. It is important not toKoehler illumination is now set. It is important not tomove the condenser up or down or change the fieldmove the condenser up or down or change the fielddiaphragm.diaphragm.Setting the Koehler IlluminationSetting the Koehler Illumination(continued)(continued)
  10. 10. Operation of the Microscope –Operation of the Microscope –Examining SmearsExamining Smears• Put smear on stage and center it under the 10XPut smear on stage and center it under the 10Xobjectiveobjective• Adjust intensity of the light to a comfortable levelAdjust intensity of the light to a comfortable levelwith the transformerwith the transformer• Open condenser diaphragm about 70% to achieve aOpen condenser diaphragm about 70% to achieve agood balance of resolution and contrastgood balance of resolution and contrast• Adjust oculars for interpupillary distance so thatAdjust oculars for interpupillary distance so thatwhen looking with both eyes only one circle of lightwhen looking with both eyes only one circle of lightis seenis seen
  11. 11. Examining SmearsExamining Smears (continued)(continued)• Adjust sharpness of image by moving adjustmentAdjust sharpness of image by moving adjustmentring on adjustable ocularring on adjustable ocular• Once 10X focus is achieved, rotate nosepiece so thatOnce 10X focus is achieved, rotate nosepiece so thatthe 40X objective is in placethe 40X objective is in place• Readjust the intensity of light to a comfortable levelReadjust the intensity of light to a comfortable levelusing the transformerusing the transformer• Use the fine adjustment knob to focus up and downUse the fine adjustment knob to focus up and downthrough the different planes of the fieldthrough the different planes of the field
  12. 12. Microscope Problems – Troubleshooting 1Microscope Problems – Troubleshooting 1ProblemProblem: Black Field: Black FieldPossible Causes:Possible Causes:• Microscope not plugged inMicroscope not plugged in• Power not available at outletPower not available at outlet• Illuminator not turned onIlluminator not turned on• Bulb burned outBulb burned out• Objective not clicked into placeObjective not clicked into place• Condenser too low withCondenser too low withdiaphragms closeddiaphragms closed
  13. 13. Microscope Problems – Troubleshooting 2Microscope Problems – Troubleshooting 2ProblemProblem: Field only partially illuminated: Field only partially illuminatedPossible Causes:Possible Causes:• Objective not clicked into positionObjective not clicked into position• Condenser not centered correctlyCondenser not centered correctly• Condenser too lowCondenser too low• Field diaphragms closed too muchField diaphragms closed too much
  14. 14. ProblemProblem: Difficulty focusing with 10X: Difficulty focusing with 10XobjectiveobjectivePossible Causes:Possible Causes:• Wrong objective in placeWrong objective in place• Objective not screwed into placeObjective not screwed into place• Not in correct plane of focusNot in correct plane of focusMicroscope Problems – Troubleshooting 3Microscope Problems – Troubleshooting 3
  15. 15. Microscope Problems – Troubleshooting 4Microscope Problems – Troubleshooting 4ProblemProblem: Difficulty focusing with 40X: Difficulty focusing with 40XobjectiveobjectivePossible Causes:Possible Causes:• Not in correct plane of focusNot in correct plane of focus• Not initially focused at 10XNot initially focused at 10X
  16. 16. Microscope Problems – Troubleshooting 5Microscope Problems – Troubleshooting 5ProblemProblem: Blurry image at 10X or 40X: Blurry image at 10X or 40XPossible Causes:Possible Causes:• Dirty objectiveDirty objective• Dirty slideDirty slide• Dirty coverslipDirty coverslipProblemProblem: Ground glass appearance: Ground glass appearancePossible Causes:Possible Causes:• Condenser too highCondenser too high
  17. 17. CreditsCreditsThanks to STD/HIV Prevention Training Center ofNew Englandin Collaboration withState Laboratory InstituteandNational Laboratory Training Network, CDC

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