Platelet aggregation: the use of optical density fluctuations to study microaggregate formation in platelet suspension.

A new approach to the investigation of microaggregate formation kinetics in platelet suspension is described. The method is based on the analysis of light transmission fluctuations produced by the changes in the number of platelets in optical channel. The relative dispersion of such fluctuations (D) was used as a parameter to estimate the degree of platelet aggregation and to analyze the aggregation kinetics.

Dual-phase response of bovine pulmonary artery endothelial cells to agonists which increase free cytoplasmic calcium concentration.

The regulation of free cytoplasmic calcium concentration ([Ca2+]i) was studied in bovine pulmonary artery endothelial cells (BPAEC). The cells were seeded on the inner surface of glass cuvettes, grown to confluency and loaded with INDO-1. Using a multiwavelength method for estimation of [Ca2+]i it was shown that in Ca2+ containing medium a rapid rise of [Ca2+]i occurs in response to bradykinin, ATP or thrombin followed by a much slower decrease in free cytoplasmic calcium.

Evaluation of the ability of intact platelets to accumulate acridine orange.

A new approach to the evaluation of the uptake of fluorescent probes by intact cells is described. Acridine orange (AO) was used because it can be selectively accumulated by serotonin-containing granules of platelets. Analysis of the fluorescence signal allows the estimation of the relative volume of the granules and the equilibrium coefficients for AO transport across the cytoplasm and granule membranes.

Accumulation and release of acridine derivatives by intact platelets.

A new approach to the investigation of accumulation of acridine derivatives and release by platelets is proposed. At high concentrations of acridine orange (AO) inside dense bodies, a two component fluorescence spectrum with maxima at 530 and 640 nm has been recorded. Computer deconvolution of spectra provided additional information on the mechanism of dye accumulation and release.