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Fluorescent Imaging

1. Align the microscope for Koehler Illumination as described above.

2. Locate your sample using either brightfield or Nomarski DIC imaging.

3. In most cases, the fluorescence signal is not bright enough to use camera live imaging to

focus. Thus it is important that what you see through the eyepieces is the same as is seen by

the camera. Do this as follows:

a. Focus on the line pattern in the right eyepiece with your right eye. Adjust the

eyepiece so that this looks like 4 double lines.

b. Then pick a feature on your slide and bring the image into focus with the right eye.

c. Now adjust the left eyepiece so that the feature is still in focus with the left eye.

d. You should now have both eyepieces adjusted so that the focus through the eyepieces

is the same as that of the image taken with the camera.