Skip to content
Finkelstein Lab Wiki

Differential Interference Contrast (DIC) Microscopy

A guide to Differential Interference Contrast (DIC) Microscopy

Differential Interference Contrast (DIC) is a microscopy technique that introduces contrast (3D effect) to images of specimens which have little or no contrast when viewed using traditional brightfield microscopy.

Contents

Section titled “Contents”
  1. Components
  2. Installing DIC

1. Components

Section titled “1. Components”

Before installing the DIC, make sure you are familiar with all the components. There are four components installed into the light path of Microscopy 3.

  1. The polarizer - changes the unpolarized light to the polarized light. It is on the right side of one slide holder (TC-C-DICP) of the head of the microscope.
  2. The first Nomarski-modified Wollaston prism - separate the incoming light into perpendicular polarized light. It, labeled with DIC N2, is on a rotatable mount of the head of the Microscopy.
  3. The second Nomarski-modified Wollaston prism – recombines the incoming perpendicular polarized light into one polarized light. We have two prisms, one (60XI) is used for 60X air objective, the other (60XIV) is for 60X water immersion objective. They are kept in the Optical box.
  4. The analyzer (the second polarizer) – removes the directly transmitted light and select the polarized light created by the second prism. It, labeled with DIC analyzer, is in the Turret 2, position 5.

2. Installing DIC

Section titled “2. Installing DIC”

The steps to install DIC:

  1. Insert the polarizer into the bright light path by pushing the TC-C-DICP holder to the left.
  2. Insert the first Nomarski prism into the bright light path by rotating the rotatable mount from A position to N2 position.
  3. Insert the second Nomarski prism (60XIV or 60XI) into the slot underneath the objective.
  4. Rotate the analyzer cube (DIC analyzer) in the Turret2 to the light path and make sure the cube in the turret1 above the analyzer cube is empty.
  5. Put your specimen on the stage. Find the focal plane.
  6. Shrink the size of aperture labeled with F on the head by turning it to the left. Lower the position of the head of the microscopy by rotating the two big black screws located at the sides of the head. Adjust the head position and get a sharp image of the aperture on the camera. If the position of the aperture is not in the center of the field of view, you can slightly turn the two silver screws between the slider TC-C-DICP and N2 rotatable mount. Fully open the aperture. The purpose of this step is to get the best contrast of the images.
  7. The DIC is ready to use.

After finishing your experiment, please put all components back to the original locations.