title: Real-time dynamics of label-free single mast cell granules revealed by differential interference contrast microscopy.
abstract: 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.
authors.affiliation: Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, Ames, IA 50011, USA.
authors.forename: Hung-Wing
authors.initials: HW
authors.lastname: Li
authors.forename: Michael
authors.initials: M
authors.lastname: McCloskey
authors.forename: Yan
authors.initials: Y
authors.lastname: He
authors.forename: Edward S
authors.initials: ES
authors.lastname: Yeung
mesh_terms: 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
chemical_list: D003208:Concanavalin A; D007073:Immunoglobulin E; D011112:Polymyxin B