|ZFIN ID: ZDB-PUB-060403-7|
Purification of hearts from zebrafish embryos
Burns, C.G., and MacRae, C.A.
|Source:||Biotechniques 40(3): 274, 276, 278 (Journal)|
|Registered Authors:||Burns (Erter), Caroline, MacRae, Calum A.|
|PubMed:||16568816 Full text @ Biotechniques|
Burns, C.G., and MacRae, C.A. (2006) Purification of hearts from zebrafish embryos. Biotechniques. 40(3):274, 276, 278.
ABSTRACTObtaining 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 also gentle enough that intact organs were obtained that, when warmed to room temperature in media, demonstrated spontaneous rhythmic contractions. Further proof of the purity of the extract and success of the technique was provided through tissue-specific quantitative RT-PCR analysis.