Real-time dynamics of label-free single mast cell granules revealed by differential interference contrast microscopy.
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.
Li
Hung-Wing
HW
Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, Ames, IA 50011, USA.
McCloskey
Michael
M
He
Yan
Y
Yeung
Edward S
ES
D000818:Animals; D015550:Cell Degranulation / Q000502:physiology*; D003208:Concanavalin A / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000502:physiology; D005260:Female; D007073:Immunoglobulin E / Q000494:pharmacology; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008561:Membrane Fusion / Q000502:physiology; D008857:Microscopy, Interference / Q000379:methods*; D011112:Polymyxin B / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar
D003208:Concanavalin A; D007073:Immunoglobulin E; D011112:Polymyxin B