PUBLICATION
Zebrafish thrombocyte aggregation by whole blood aggregometry and flow cytometry
- Authors
- Sundaramoorthi, H., Panapakam, R., Jagadeeswaran, P.
- ID
- ZDB-PUB-150423-2
- Date
- 2015
- Source
- Platelets 26(7): 613-9 (Journal)
- Registered Authors
- Jagadeeswaran, Pudur
- Keywords
- Aggregometry, flow cytometry, thrombocyte, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Arachidonic Acid/pharmacology
- Blood Platelets/drug effects
- Blood Platelets/metabolism*
- Flow Cytometry*/methods
- Kinetics
- Platelet Aggregation*/drug effects
- Platelet Function Tests*
- Zebrafish/blood*
- PubMed
- 25902147 Full text @ Platelets
Citation
Sundaramoorthi, H., Panapakam, R., Jagadeeswaran, P. (2015) Zebrafish thrombocyte aggregation by whole blood aggregometry and flow cytometry. Platelets. 26(7):613-9.
Abstract
Zebrafish has become an excellent model system to study mammalian hemostasis. Despite our extensive efforts to develop technologies to measure zebrafish hemostasis and even with previously established thrombocyte qualitative and quantitative functional assays, quantifying thrombocyte function for high throughput applications has been a challenge. In this paper, we have developed two quantitative methods to estimate thrombocyte aggregation: one by whole blood aggregometry and the other by flow cytometry. We found that it is possible to conduct whole blood aggregometry using only 2 µl of blood and the currently available aggregometer. Each of three agonists, arachidonic acid, ADP, and collagen yielded impedance curves similar to those obtained with human blood. We were also able to use flow cytometry to indirectly quantify the extent of thrombocyte aggregation by labeling whole blood with mepacrine, aggregating in the presence of each of the above agonists, separating the aggregates from the white blood cells by centrifugation, and then sorting the resulting white cell fraction for thrombocyte numbers. These methods have high throughput capabilities and have the potential to be used in large scale screens to detect and characterize mutants with thrombocyte functional defects or to identify genes involved in thrombocyte function by large scale knockdowns.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping