Practical Microscopy.

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Presentation transcript:

Practical Microscopy

Size matters:How large are cells? Let's put things into perspective....

Phospholipid membrane Organism Size Prokaryotic cell 1 – 10 μm Animal Cell 10 – 30 μm Plant Cell 10 – 100 μm Virus < 100 ηm DNA molecule 2 ηm Phospholipid membrane 10 ηm

What can you visualise with the naked eye?

What can you visualise with the naked eye? 0.1 mm (100 μm) .

Magnification X400 – X1500

Magnification Electron microscope: X 1500 – X500,000 (and incredible detail [resolution])

Magnification: Scanning Tunnelling microscope Resolution: 0.1 ηm: can show atom – atom!

Biggest and smallest cells Human egg: 130 μm Human sperm: 60 x 5 μm Some neurons can be 1 m long!

You should be able to identify Eye piece (X 10 magnification) Light source Objective lenses (X 4, X10, X40) Coarse Adjustment knob Fine adjustment knob

Body tube Eye Piece Arm 4x Objective 10x Objective Stage 40x Objective Revolving nosepiece Arm 4x Objective 10x Objective Stage 40x Objective Stage Clips Coarse Adjustment Diaphragm Fine Adjustment Light source Base

Guidelines for working with microscopes ALWAYS carry a microscope with BOTH hands The Three-Step Method 1. Use the scanning objective (red X 4) and bring it into focus using the LARGE KNOB (coarse adjustment) 2. Switch to low power (yellow X 10) and refocus using the SMALL KNOB – re-centre if necessary. 3. Switch to high power (blue X 40) and ONLY focus with the small knob (fine adjustment)

Calculating magnification/image size/actual size The ‘I am’ triangle

Clean Up, Clean Up, Everybody Clean Up! When you are finished with the microscopes, should store them appropriately: Set to the lowest objective (4X). Light intensity to lowest setting. Power off. Mechanical stage set such that the mechanism does not poke a hole through the custom covers Stage down Cord wrapped (gently around the scope is sufficient - nothing fancy). Cover in place Of course, the stage should be free from dirt, water, and solutions. However, you should not attempt to clean the optics. Report dirty oculars or objectives

What are you looking at? Animal cells with/without methylene blue Plant cells (onion and elodea) with/ without methylene blue Plant cells with/without iodine Protists (paramecium sample) Protists (pond water)

Homework for Week 3 Organelle identification Practice questions on organelle size Practice questions on magnification Practice questions on cell organelles (you will have a quiz on this next week!)