Whole-mount in situ hybridization of small invertebrate embryos using laboratory mini-columns.
To facilitate the handling of small invertebrate embryos during whole-mount in situ hybridization, a method of solution exchange using laboratory mini-columns was developed. This protocol speeds time-consuming aspiration of buffers, and avoids accidental loss of the embryos, by gently pushing solutions through the column using air pressure from a syringe after each incubation. The next buffer is then added using a pipettor. Embryos are retained on a filter within the column. As many columns as desired may be processed in parallel for different probes or stages.
Irvine
Steven Q
SQ
Biological Sciences Center, University of Rhode Island, Kingston, RI 02881, USA. steven.irvine@uri.edu
D000818:Animals; D004330:Drosophila; D004622:Embryo, Mammalian; D004625:Embryo, Nonmammalian; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods* / Q000592:standards*; D051379:Mice; D015203:Reproducibility of Results; D013594:Syringes