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Study of calcium signaling dynamics in single platelets
ISTH Academy. Spiryova D. Jul 9, 2019; 264758; PB1569
Ms. Daria Spiryova
Ms. Daria Spiryova
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PB1569

Study of Calcium Signaling Dynamics in Single Platelets

D. Spiryova
Novosibirsk State University, Physics Department, Novosibirsk, Russian Federation

Main Topic: Platelets and Megakaryocytes
Category: Platelet Signaling

Background: Platelets are blood cells that are the main component of hemostasis. Platelet activation should be actively studied, as today it is important for the diagnosis of diseases associated with the problems of blood clotting. To understand the basic mechanisms and functions of platelets, it is necessary to study in vitro platelet responses to various stimuli, such as artificial activation of a single platelet.
Aims: The purpose of this study is to investigate the release of calcium in floating platelets.
Methods: Calcium indicator Fluo-4 was added to the cell suspension and the sample was placed under a fluorescent microscope. We observed some cells attached to the surface and others floating higher. Activation of platelets was carried out traditionally, adding ADP or adrenaline, or optically, using a photolabile caged analogue of epinephrine, which is released after exposure to radiation of a certain wavelength. We were able to get the fluorescence intensity of floating cells by tracking the movement of each individual platelet using the ImageJ TrackMate plugin.
Results: The fluorescence dynamics of the calcium indicator was recorded at a rate of 3-frame per second. Measurements showed the release of calcium in floating cells. In addition, we have shown that the photolabile caged analogue of epinephrine can be used to activate platelets optically, which will significantly reduce the measurement error associated with the displacement of cells after the addition of conventional adrenaline. We were able to activate particular cells in a microscope slide using translating optical fiber.
Conclusions: The results formed the basis of a new method of studying the activation of single platelets. In addition, the study showed differences in the behavior of attached and floating cells, such as fluorescence intensity.

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