Protocol

Differential Interference Contrast (DIC) Imaging of Living Cells

This protocol was adapted from "A Practical Guide: Differential Interference Contrast Imaging of Living Cells," Chapter 4, in Imaging in Neuroscience and Development (eds. Yuste and Konnerth). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.

INTRODUCTION

This protocol describes the use of electronically enhanced, differential interference contrast (DIC) imaging techniques for the study of living cells. Methods for DIC imaging of cells using an inverted microscope and for infrared (IR) imaging of brains slices using an upright microscope are presented. DIC microscopy (also known as Nomarski microscopy) is extremely useful for resolving individual cells and cellular organelles in live, unstained tissue. The data obtained are valuable in their own right, as well as for complementing fluorescence microscopy data. DIC microscopy can be applied to cell culture, brain slices, and even intact organisms (such as embryos). High-quality images of living cells in brain slices can also be obtained using a combination of DIC optics and IR video microscopy.