Olympus IX71 microscope

 

Always set condenser for Kohler illumination.
1. Focus on sample
2. set condenser for BF
3. Close Field Diaphagm
4. Move condenser up and down using knob on right until center octagon is in focus
5. Open field diaphragm to edge of visible field, not all the way.

Set condenser for brightfield or phase contrast:

Objective Lens setting for phase Condenser setting for brightfield
4X Postion 1: PhL Position 5: BF
10X Position 2: Ph1 Position 5: BF
20X

Position 2: Ph1

Position 5: BF
40X Position 3: Ph2 Position 5: BF

 

Important for best image quality:

The 20X and 40X lenses have adjustable collars. You turn these rings to optimize the lens for the type of chamber you are imaging. A number 1.5 coverslip needs the ring adjusted to the 0.17 position. A plastic flask or dish will need a setting in the 1 to 2 range.

However, you cannot easily see the collar underneath the stage. Therefore:
1. Focus on sample best possible.
2. Reach under stage to turn ring on objective a little.
3. Refocus, turn ring, and repeat until image has maximum crispness.

Also, this may also be done when in the fluorescence mode. It is important to make this adjustment or else your pictures will not be in proper focus.

 

Section to be written: fluorescence microscopy settings and how to take pictures in linear range for quantification.

 

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comments, questions, suggestions for this web page: Michael.Cammer@med.nyu.edu or mcammer@gmail.com