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Practical:2 L ight Microscopy

Practical:2 L ight Microscopy. presented by: Ms Asmaa A Basonbul. Objectives. Recognize different parts of a compound light microscope. Learn how using appropriate objective lenses e.g 10x for focusing 40x for wet preparation 100x oil immersion

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Practical:2 L ight Microscopy

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  1. Practical:2Light Microscopy presented by: Ms Asmaa A Basonbul

  2. Objectives • Recognize different parts of a compound light microscope. • Learn how using appropriate objective lenses e.g 10x for focusing 40x for wet preparation 100x oil immersion • Learn how using condenser and iris diaphragm to adjust the light source to the optimum.

  3. Introduction Microscopy: The use of microscopes in all their various forms. Q. Why we use microscope in haematology lab? To study various blood cells and inclusion bodies.

  4. How? By using a system of lenses and illumination sources it’s make object visible. Q. How many times the microscope can magnify that object? Microscope can magnify an object from 100-1000 times of it’s original size.

  5. Principle To magnify the object the LM use a system of lenses (objectives and oculars) to manage the path of light beam that travel b/w object which we’re studied to the eye.

  6. Parts of light microscope • Eye picec (ocular lenses): Magnify lens with magnification power 10x.

  7. Cont: Parts of light microscope • Body tube: Contains mirrors and prisms that transmit the image from the objective lens to the ocular lens.

  8. Cont: Parts of light microscope • Objective lenses: Primary lenses that magnify specimen • Low power 10x • High power 40x • Oil immersion 100x

  9. Cont: Parts of light microscope • Stage: Holds slide in position.

  10. Cont: Parts of light microscope • Condenser: Lens system that condenses light before it passes through the specimen.

  11. Cont: Parts of light microscope • Iris diaphragm: Control the amount of light entering the condenser.

  12. Cont: Parts of light microscope • Coarse and fine adjustment knobs: Used for focusing the specimen.

  13. Cont: Parts of light microscope • Light: Source of illumination, bulb.

  14. Types of Light Microscopy • Bright field microscopy. • Dark field microscopy. • Fluorescence microscopy. • Phase contrast microscopy.

  15. Bright filed microscope Light bulb Day light transfer bent Object in the specimen

  16. This method use to examine stained preparation

  17. Dark filed microscope Special objective U Special condenser in mico Light source transfer bent Object in the specimen “Microorganism”

  18. This method use to examine unstained preparation

  19. Fluorescence microscope

  20. Fluorescence microscope

  21. Specimen labeled with fluorescence dye • Acridine orange • Auramine/rhodamine

  22. Specimen labeled with fluorescence dye • Calcofluor white

  23. Phase contrast microscope contrast Bright(denser) Dark(lighter) special objectives Light source Picture appear with different darkness and brightness bent transfer Object in the specimen Special condenser

  24. This method use to examine unstained preparation

  25. Materials • Compound microscope. • Lens cleaning paper/ cloth. • Immersion oil. • Wet preparation. • Stained preparation.

  26. Wet preparation method

  27. Oil immersion objective(100x): • Put drop of oil immersion on the slide. • Turn the oil immersion objective 1oox and put above the slide. • The 100x objective will immersed in the oil. • The picture is transmitted from objective lens to ocular lens.

  28. The oil helps to keep light rays together as they pass b/w the specimen and objective lens.

  29. Stained preparation method

  30. Total magnification power of a microscope: = power of the objective lens X power magnification of eye piece e.g : using oil immersion objective 100x ocular lens (eye piece) 10x ??!! So 100x X 10x =1000

  31. Resolving power of microscope: Is measure of its ability to discriminate b/w two adjacent objects. Absolute limit of the resolving power = wave length of LM = 400-800nm

  32. Maintenance of microscope • The microscope is delicate instrument which is must be: • Handle gently. • Put in a clean environment away from chemical, direct sunlight, evaporation, heating source or moisture. • Cleaned immediately if the stage is contaminated with saline to avoid corrosion. • The temperate climate humidity and high temperature causes growth of fungus which can damage optical surfaces

  33. Cleaning the microscope • Optics: • The low and high power lens paper tissue. • Oil immersion lens paper tissue and few drop from xyelen (if needed). • Non-optics: • Eyepices: Using the soft camel-hair brush to remove the dust from in and out side.

  34. Cleaning the microscope • Condenser and iris diaphragm: Using soft cloth or tissue moistened with toluene and the mirror with 5% alcohole. Other parts cleaned with mild detergent and remove grease or oil with petroleum ether followed by 45% ethanol in water.

  35. Q. Defined the different parts of microscope?

  36. Thanks for listening Let us to start practical part

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