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Adjusting the microscope for Nomarski DIC imaging

1) Select DIC grayscale or DIC color on the lower left of the SPOT camera program window.

2) Make sure the camera filter selector is pushed in for grayscale or pulled out for color.

3) Insert and, using the LIVE mode on the camera, focus on a sample using the objective you

intend to use for your imaging.

4) Adjust the lower DIC filter to the extreme right position.

5) Move the condenser to the correct position to match the objective you plan to use.

DICL = 10x or 20x

DICM = 40x

DICH = 100x (oil immersion)

6) Insert the upper DIC filter. Adjust the microscope light using the neutral density filters.

7) Adjust the position of the lower DIC filter until you have the amount of DIC contrast you

want.

Adjust C ondenser t o

desired D IC s etting

8) Remove your slide and select "camera > Get flatfield" from the camera menu. Make a

flatfield and save it as "ff". Overwrite the previous flatfield. This procedure takes an image

of your light path through the microscope and will subtract it from your image. This will

remove any uneven light from the final image. A new flatfield must be made if you change

anything in your setup such as objective or position of lower

DIC filter.

9) Insert your slide and proceed with locating and focusing

your sample.

Upper D IC f ilter i s

out f or B F a nd

Fluor. I maging a nd

in f or D IC i maging.

Lower D IC f ilter i s s et f or

desired a mount o f

contrast.

Turn f rom l eft t o r ight