Microscope (basic use)

The objectives currently on the scope are fine for most uses (ie looking at cells). However, for finer images, there are higher quality objectives for phase and fluorescence in Michael's office.

If you need to clean the objectives, eyepieces, or any other lense-like surface, use lense paper or a q-tip with methanol. If you use immersion oil, make sure you clean it off the objective and surrounding areas with lense paper and methanol.

Brightfield Imaging:
-Plain light microscopy. Ok for looking at cells, but not as good as phase or nomarsky

  • Set objective to lowest magnification (10X) and place slide on stage.
  • Turn light on to about half power (wheel on front base of scope).
  • Disengage dichroic filters by pulling the Mirror Block Selector Knobs (located on both sides of scope above the objectives) out. (
  • Make sure that the anlyzer is not in place by pulling the black knob directly above the objective out.
  • Turn filter wheel (located under the stage) to 0 setting
  • Focus using knob on left or right of scope
  • Adjust light by moving condenser (under stage) or changing diaphragm (under stage).

Phase Imaging:
-Best for looking at cells, morphology is most clear.

  • Set objective to lowest magnification phase objective (marked with Ph) and place slide on stage.
  • Turn light on to about half power (wheel on front base of scope).
  • Disengage dichroic filters by pulling the Mirror Block Selector Knobs (located on both sides of scope above the objectives) out. (
  • Make sure that the anlyzer is not in place by pulling the black knob directly above the objective out.
  • Set filter wheel (located under the stage) to Ph setting (1 or 3) with number that matches the number on the objective you are using (use Ph3 for Ph2 objectives).
  • Focus using knob on left or right of scope
  • Adjust light by moving condenser (under stage) or changing diaphragm (under stage).

Nomarski Imaging:
-Produces three-dimensional image. Good for looking at cells.

  • Set objective to lowest magnification (10X) and place slide on stage.
  • Turn light on to about half power (wheel on front base of scope).
  • Disengage dichroic filters by pulling the Mirror Block Selector Knobs (located on both sides of scope above the objectives) out. (
  • Focus using knob on left or right of scope
  • Turn filter wheel (located under the stage) to numerical setting that matches the objective you are using (ie, 20, 40, 100).
  • Make sure that the polarizer is in place (in the condenser, below the stage)
  • Make sure that the anlyzer is place by pushing the black knob directly above the objective in.
  • Adjust prism (black rectangular device located above objectives) by moving it in and out and by turning the silver knob on its end. As you turn the knob, the degree of three-dimensionality should change.
  • Adjust light by moving condenser (under stage) or changing diaphragm (under stage).

Fluorescence imaging:
-Use with mercury lamp. Can be done in both Nomarski and Bright Field settings.

  • Turn on lamp 5-10 minutes before using to allow time to warm up.
  • Always leave the lamp on for at least 10 minutes after turning on (ie, don't turn on and off quickly) and wait 10 minutes after turning off before turning on again to save the bulb.
  • Use settings as for bright field or Nomarski.
  • Make sure that a dichroic filter cube with the appropriate filter (ie, DAPI, GFP, etc) is in place.
  • After focusing, turn off white light, and lower condenser (glass in condenser can reflect light and diminish image quality).
  • Make sure that the shutter and UV diaphragm are fully open.