Simple and rapid quantification of thrombocytes in zebrafish larvae

Zebrafish
Michael C Huarng, Jordan A Shavit

Abstract

Platelets are a critical component of hemostasis, with disorders of number or function resulting in coagulation disturbances. Insights into these processes have primarily been realized through studies using mammalian models or tissues. Increasingly, zebrafish embryos and larvae have been used to study the protein and cellular components of hemostasis and thrombosis, including the thrombocyte, a nucleated platelet analog. However, investigations of thrombocytes have been somewhat limited due to lack of a robust and simple methodology for quantitation, an important component of platelet studies in mammals. Using video capture, we have devised an assay that produces a rapid, reproducible, and precise measurement of thrombocyte number in zebrafish larvae by counting fluorescently tagged cells. Averaging 1000 frames, we were able to subtract background fluorescence, thus limiting assessment to circulating thrombocytes. This method facilitated rapid assessment of relative thrombocyte counts in a population of 372 zebrafish larvae by a single operator in less than 3 days. This technique requires basic microscopy equipment and rudimentary programming, lends itself to high throughput analysis, and will enhance future studies of thrombop...Continue Reading

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Citations

Dec 5, 2019·Journal of Experimental Zoology. Part B, Molecular and Developmental Evolution·Gerald Soslau
Aug 21, 2021·Bioinformatics and Biology Insights·Samrat Thapa, David L Stachura
Sep 26, 2021·Scientific Reports·Simone MorrisStefan H Oehlers

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Methods Mentioned

BETA
light microscopy
blood collection
transgenic

Software Mentioned

GraphPad
Adobe AfterEffects
NeatVideo
HalfShutter
ImageJ
Excel

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