Real-time dynamics of label-free single mast cell granules revealed by differential interference contrast microscopy

Li, Hung-Wing; McCloskey, Michael; He, Yan; Yeung, Edward S.
January 2007
Analytical & Bioanalytical Chemistry;Jan2007, Vol. 387 Issue 1, p63
Academic Journal
We demonstrate the capability of differential interference contrast (DIC) microscopy as a simple and useful tool for studying cellular events without fluorescence labeling. By coupling an advanced DIC microscope to a computer-controlled motorized vertical stage and a high-speed, high-resolution CCD camera, real-time three-dimensional monitoring is possible in a high-throughput manner. The performance among three modes of microscopy, bright-field, dark-field and DIC, in terms of horizontal resolving power and vertical sectioning was investigated. As a model, exocytosis of rat peritoneal mast cells was recorded on the subsecond time scale. Three-dimensional tracking of granules during degranulation was achieved and granule–granule fusion before plasma membrane fusion was recorded.


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