Pierce Lain X, Harrison David, Liang Jennifer Ostrom
Department of Genetics, Case Western Reserve University, Cleveland, Ohio 44106-7080, USA.
Zebrafish. 2007 Fall;4(3):169-77. doi: 10.1089/zeb.2007.0511.
Patterning of zebrafish and other vertebrate embryos proceeds according to consistent, predictable developmental time courses. Because zebrafish spawn primarily during the first few hours after dawn, many important developmental stages typically occur during the middle of the night. As an automatic, accurate way to fix embryos at these inconvenient times, we have developed the Time Reaper 5-Channel Automatic Liquid Dispenser (TimeR). The TimeR delivers up to 50 mL of liquid to embryos in a Petri dish at preset times. We have used the TimeR to deliver paraformaldehyde and fix zebrafish embryos at different stages of development. We find that the pattern of expression for a number of genes is indistinguishable between embryos fixed manually and with the TimeR. The TimeR is also suitable for fixing embryos for whole-mount immunostaining, but care needs to be taken to find conditions that preserve the antibody's epitope. The TimeR is inexpensive to make, and can be constructed using tools present in most machine shops. In addition to fixing embryos, the TimeR will be useful for any experiment that requires automatic delivery of milliliter amounts of liquid.
斑马鱼和其他脊椎动物胚胎的模式形成是按照一致、可预测的发育时间进程进行的。由于斑马鱼主要在黎明后的最初几个小时内产卵,许多重要的发育阶段通常发生在午夜时分。作为一种在这些不便的时间自动、准确地固定胚胎的方法,我们开发了时间收割者5通道自动液体分配器(TimeR)。TimeR可在预设时间向培养皿中的胚胎输送多达50毫升的液体。我们已经使用TimeR输送多聚甲醛并固定处于不同发育阶段的斑马鱼胚胎。我们发现,许多基因的表达模式在手动固定的胚胎和使用TimeR固定的胚胎之间没有区别。TimeR也适用于固定胚胎以进行整体免疫染色,但需要注意找到能保留抗体表位的条件。TimeR制作成本低廉,可以使用大多数机械车间现有的工具来构建。除了固定胚胎外,TimeR对于任何需要自动输送毫升量液体的实验都将是有用的。