CTAC ICBR SOP: Nomarski (DIC) Imaging Using the Leica TCS SP5 Confocal Microscope

Materials Required:

  • LCS software
  • Computer
  • Leica TCS SP5 Confocal Microscope

Purpose:

To facilitate operation of the Leica TCS SP5 Confocal Microscope in imaging sample(s) using Nomarski Imaging.

 

Nomarski (DIC) Imaging

Nomarski or Differential Interface Contrast imaging is a light microscopy technique that will enable you to see a bright field image of your sample with some contrast of depth depending on the thickness of your sample.  This is very useful to see where your cells actually are in relationship to the fluorescent tags that you are imaging with the confocal scanner.

To view a DIC image in either single image or sequential scanning format click on

Additional Channels Button

on the left hand screen and choose “Scan-DIC”

PMT Box

 

 

 

 

 

 

 

 

This will open up an additional window alongside the color windows that you are using for your confocal images.  This image can then be adjusted for “Gain” and “Offset” in the standard fashion.

 

IMPORTANT:  When you are finished clean the objectives used with clean lens tissue paper.