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If your image contrast is bad, you may not be using the right type of microscopy.
If your image contrast is bad, you may not be using the right type of microscopy.
Usually, visible light microscopes are set to brightfield mode, which is usually ideal for stained samples. However, when faced with a lack of staining, there are other types of microscopy shown to the right that may increase contrast.
Phase microscopy is ideal for unstained specimens 5-10 micrometers thick above the nucleus and less than 1 micrometer thick on the cell's periphery.
Step 1: Set up Kohler illumination (see here for more information)
Step 2: Install phase rings in condenser
Step 3: Remove eyepieces and replace with a phase centering one
Step 4: Place into focus, so that phase plate and phase ring are visible
Step 5: Turn on the lowest magnification and the corresponding phase annulus
Step 6: Using the adjustment screws on the condenser, move the phase plate so that it aligns with the phase ring
Step 7: Repeat steps 4-6 for other objectives
Step 8: Replace phase centering eyepieces with normal eyepieces
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