SUMMARY
Differential interference contrast (DIC) microscopy has been widely used for many years. However, many biologists employ DIC inefficiently and frequently misinterpret the resulting images. In an effort to improve the standard of use, a method of simulating DIC microscopy on the cathode‐ray tube (CRT) and x–y plotter of a computer was therefore developed. This paper describes the program and gives examples of its application, emphasizing some common errors of operation of the DIC microscope and interpretation of the image.
Simulations are made for objects of negligible thickness, and in one azimuth of the microscope at a time. The method shows clearly the profiles of the phases and amplitudes of the interfering waves obtained for a variety of objects and demonstrates unequivocally the kinds of images obtained. It is possible to show very quickly how the image changes as instrumental parameters, such as phase‐bias, amplitude‐ratio, extinction factor, and beam‐splitter shear are altered.
Simulation was found useful as a tool to learn DICM and as an aid to the interpretation of complicated images obtained with real objects. Examples of common errors in adjusting the microscope and interpreting images are given in this paper.
The simulation program was developed for the computer of the Denver Universal Microspectroradiometer (David & Galbraith, 1975; Galbraith et al., 1975), but it can be used on a number of compatible computing systems. The program construction is explained to enable comparable programs to be written for different computers.