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

TitleReal-time dynamics of label-free single mast cell granules revealed by differential interference contrast microscopy
Publication TypeJournal Article
Year of Publication2007
AuthorsLi HW, McCloskey M, He Y, Yeung ES
Journal TitleAnalytical and Bioanalytical Chemistry
Volume387
Pages63-69
Date PublishedJan
Type of ArticleArticle
ISBN Number1618-2642
Accession NumberISI:000242968000015
KeywordsBETA-CELLS, CAPACITANCE MEASUREMENTS, CCD camera, CHROMAFFIN GRANULES, EOSINOPHILS, FUSION, imaging, mast cells, membrane, microscopy, REGULATED EXOCYTOSIS, SECRETORY GRANULES, TRACKING, VESICLE
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.

DOI10.1007/s00216-006-0403-8
Alternate JournalAnal. Bioanal. Chem.