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Zebrafish thrombocyte aggregation by whole blood aggregometry and flow cytometry
Hemalatha Sundaramoorthi1, Rekha Panapakam, Pudur Jagadeeswaran
1a Department of Biological Sciences , University of North Texas , Denton , TX , USA.
Platelets
|April 23, 2015
Summary
Researchers developed two high-throughput methods to quantify zebrafish thrombocyte aggregation using whole blood aggregometry and flow cytometry. These assays enable large-scale screening for thrombocyte function defects and gene identification.
Area of Science:
- * Zebrafish hematology and hemostasis research.
- * Development of high-throughput screening assays.
Background:
- * Zebrafish are a valuable model for studying mammalian hemostasis.
- * Quantifying zebrafish thrombocyte function for high-throughput applications remains challenging.
Purpose of the Study:
- * To develop and validate two novel quantitative methods for assessing zebrafish thrombocyte aggregation.
- * To enable high-throughput screening for thrombocyte functional defects and gene identification.
Main Methods:
- * Whole blood aggregometry using minimal blood volume (2 µl) and standard aggregometers.
- * Flow cytometry-based indirect quantification of thrombocyte aggregation via mepacrine labeling and white blood cell sorting.
Main Results:
- * Whole blood aggregometry successfully measured thrombocyte aggregation in response to arachidonic acid, ADP, and collagen.
- * Flow cytometry provided indirect quantification of thrombocyte aggregation, correlating with agonist stimulation.
- * Both methods demonstrated high-throughput capabilities suitable for large-scale screening.
Conclusions:
- * Developed quantitative assays for zebrafish thrombocyte aggregation using aggregometry and flow cytometry.
- * These methods facilitate efficient screening for thrombocyte functional mutants and gene discovery.
- * Advances in zebrafish hemostasis research are enabled by these high-throughput techniques.
