Nikon Fluorescent Microscope

 

Set-Up

1.      Microscope and power supply plugged in and turned on (done by Instructor or T.A.).

2.      Top filter out.  Viewing through no filter.

3.      Mid-level filter in center.  Viewing without phase contrast.

4.      Aperture lever far to right.  Aperture is open (O).

5.      Light intensity mid-range.

6.      Magnification 10x or 20x.

7.      Fluorescent filter cube options:

·        TRITC Filter: see Ethidium.

Left metal lever pushed in. View through TRITC (blue) cube on left-hand side.

·        FITC Filter: see FITC and Calcein.

Right metal lever pushed in. View through FITC (green) cube on right-hand side.

·        Both metal levers out.  View through neither cube, which is clear brightfield.

8.      Fluorescent light pass options:

·        Black lever pushed in far (to left) to block fluorescent light for brightfield viewing.

·        Black lever pulled out (to right) to let fluorescent light pass for fluorescent applications.

 

Operation

1.      Turn on power on front of power source box (done by Instructor or T.A.).

2.      Place cells on stage.

3.      Turn off overhead lights, if needed.

DO NOT FIDDLE AROUND WITH BUTTONS NOT DESCRIBED TO GET THE MICROSCOPE TO “WORK.”  PLEASE ASK FOR HELP.

4.      Focus in on cells of interest using brightfield imaging:

·        Close fluorescent light pass.

·        Pull both metal filter cube levers out to see through neither filter cube.

·        Turn on power/light to microscope.

·        Focus on cells using gross and fine focus controls.

5.      To see through FITC filter:

·        Turn off power to microscope.

·        Push in metal lever on right-hand side (FITC).  (Left-hand side lever pulled out.)

·        Open fluorescent light pass.

6.      To see through TRITC filter:

·        Turn off power to microscope.

·        Push in metal lever on left-hand side (TRITC).  (Right-hand side lever pulled out.)

·        Open fluorescent light pass.

7.      To see through brightfield:

·        Close fluorescent light pass.

·        Pull both metal filter cube levers out to see through neither filter cube.

·        Turn on power/light to microscope.