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
References
Dec 22, 1999·British Journal of Haematology·P JagadeeswaranD Troyer
Aug 16, 2005·Blood·Hui-Feng LinRobert I Handin
Dec 18, 2007·IEEE Transactions on Pattern Analysis and Machine Intelligence·Ce LiuWilliam T Freeman
Jan 5, 2010·Blood Cells, Molecules & Diseases·Seongcheol KimPudur Jagadeeswaran
Nov 3, 2010·Blood Cells, Molecules & Diseases·Maira CarrilloPudur Jagadeeswaran
Dec 14, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Li ZhangCharles P Lin
Dec 20, 2011·Blood·Tim ThijsKatleen Broos
Feb 22, 2012·Methods in Enzymology·Felix Ellett, Graham J Lieschke
Jul 4, 2012·The Journal of Clinical Investigation·Cristina Santoriello, Leonard I Zon
Jul 11, 2012·Advances in Hematology·Gauri KhandekarPudur Jagadeeswaran
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Oct 11, 2012·Advances in Hematology·Arunima GhoshJordan A Shavit
Mar 8, 2013·Seminars in Thrombosis and Hemostasis·Natalia BunimovCatherine P M Hayward
Apr 19, 2013·Nature·Kerstin HoweDerek L Stemple
Mar 29, 2014·Circulation Research·Julia M BurkhartRené P Zahedi
Jul 24, 2014·Current Opinion in Hematology·Angela C Weyand, Jordan A Shavit
Sep 23, 2014·Zebrafish·Chereen CollymoreMichael L Kent
Citations
Dec 7, 2018·Blood Advances·Megan S RostJordan A Shavit
Jun 4, 2017·Blood·Zhilian HuJordan A Shavit
Dec 5, 2019·Journal of Experimental Zoology. Part B, Molecular and Developmental Evolution·Gerald Soslau
Jun 7, 2019·Blood Advances·Angela C WeyandJordan A Shavit
Nov 10, 2020·Blood Advances·Cristina FreireMarguerite Neerman-Arbez
Aug 21, 2021·Bioinformatics and Biology Insights·Samrat Thapa, David L Stachura
Sep 26, 2021·Scientific Reports·Simone MorrisStefan H Oehlers