Purification of Hearts from Zebrafish Embryos
Author(s) -
C. Geoffrey Burns,
Calum A. MacRae
Publication year - 2006
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112135
Subject(s) - biology , embryonic stem cell , embryo , in situ hybridization , zebrafish , transplantation , gene expression , andrology , progenitor cell , computational biology , microbiology and biotechnology , stem cell , gene , genetics , medicine
Have a Heart Obtaining pure organ preparation by microdissection from small organisms, such as the zebrafish, Dario rerio, is itself a sufficiently arduous task. Isolating such organs in the same manner from the fish in its embryonic stage is near impossible. With this technical hurdle to mount, Burns et al. developed a simple and straightforward isolation technique, described on p. 274 of this issue, which yielded good quantities of almost homogeneous zebrafish heart. The basic setup included a fine, large-gauge needle and syringe, clamped securely over a microfuge tube containing the zebrafish embryos (those between and 2 and 4 days postfertilization were used). Making use of the shear forces created by repeatedly drawing up and expelling the solution containing suspended embryos, the yolk sac could be broken, and the embryos fragmented. Filtering through two grades of nylon mesh generated a highly pure suspension of zebrafish hearts. The procedure was sufficiently vigorous to produce a good yield, but ...
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