Nikon E800 Manual de Instruções - Página 4

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1. Fluorescent Imaging

Adjusting the microscope for Koehler illumination.

1) Insert a sample and focus it using an objective that you typically use for imaging.

Tip: Focus on the edge of the coverslip to set the

course focus. Then focus up and down using fine

focus to locate sample.

2) Stop down (close) field aperture by rotating

dial on left side of microscope base. You may see a

round circle of light through the eyepieces that has

a blurry edge.

3) Adjust the condenser height (#1: black knob on left of condenser) until the circle of

light is smaller and the edges are sharp. This will appear as the octagon (8-sided) shape

of the aperture.

4) Open up the field aperture to fill ~half the area

with light.

5) Adjust the aperture centering with the two

silver knobs (#2 below the condenser turret). When

centered, open the field aperture until the entire field

is full of light.

Note: You may have to adjust the light for other

objectives. Lower magnification objectives have a

wider opening so will require opening the field

aperture more.

6) Close the condenser aperture by moving the

slider to the right. This is the position of maximum

diffraction. This will give additional contrast to the

sample but reduces resolution.

Open the condenser aperture until the light level does

not change. This is the position of maximum

resolution and minimum diffraction. Use this position

for all higher magnification objectives.

<Open......Close>

O | | | | | | | | | o

You have now aligned the microscope for routine brightfield imaging.

Field

Aperture

Condenser A perture