Fluorescence Microscopy for the Axioplan 2

Equipment:

Plan-Neofluar objectives.

Reflected light illuminator.

 

Quick Start:

  1. Identify the object with bright field, then block transmitted light.
  2. Center and focus epi-illuminating arc
  3. Select fluorescence filter cube and filter wheel setting.
  4. Set up Koehler epi-illumination
  5. Center epi-path field iris and open epi-path aperture iris

 

Step 1. Identify the object for fluorescence with bright field or DIC objectives, then block transmitted light.

 

Step 2. Center and focus epi-illuminating HBO arc source using adjustment aid.

 

Step 3. Select fluorescence filter cube and filter wheel setting appropriate to the fluorophore in the specimen.

 

Step 4. Set up Koehler epi-illumination by focusing sample, removing diffusing screen and closing down the epi-path field iris.

 

Step 5. Center epi-path field iris and open it so that it is just outside the object or field of interest. Open epi-path aperture iris to level of desired excitation light (for faint specimens, open it, or push in, all the way).