Glyoxal-based fixation of Drosophila embryos for immunofluorescence staining and RNA in situ hybridization.
Glyoxal-based fixation of Drosophila embryos for immunofluorescence staining and RNA in situ hybridization.
复制标题
DOI：
10.1016/j.xpro.2023.102385
复制
发表时间：
2023-09-15
期刊：
影响因子：
--
通讯作者：
Sokac, Anna Marie
中科院分区：
文献类型：
--
作者：
Amin, Shrunali;Basu, Malika;Buzinova, Valeria;Delgado, Anthony;Mahadevan, Tejas;Mishra, Sanya;Zaida, Sarah;Wang, Xi;Sokac, Anna Marie
双醛乙二醛是一种替代的化学固定剂，它比甲醛更快地交联组织，保持更高的抗原性，并且比甲醛或戊二醛的危害更小。在这里，我们提出了一种基于乙二醛的固定方案，用于果蝇胚胎。我们描述了制备无酸乙二醛，固定胚胎，然后用免疫荧光（IF）抗体染色的步骤。我们还描述了使用乙二醛固定胚胎进行RNA荧光原位杂交（FISH）和FISH + IF （FISH-IF）的方法。该方案改编自Bussolati等人和Richter等人的方法，适用于果蝇胚胎。乙二醛为果蝇胚胎的固定提供了甲醛的替代品本方案描述了制备无酸乙二醛和固定胚胎的步骤乙二醛固定胚胎的免疫荧光染色方法是详细的RNA原位杂交方法也描述了出版商注：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指导方针。双醛乙二醛是一种替代的化学固定剂，它比甲醛更快地交联组织，保持更高的抗原性，并且比甲醛或戊二醛的危害更小。在这里，我们提出了一种基于乙二醛的固定方案，用于果蝇胚胎。我们描述了制备无酸乙二醛，固定胚胎，然后用免疫荧光（IF）抗体染色的步骤。我们还描述了使用乙二醛固定胚胎进行RNA荧光原位杂交（FISH）和FISH + IF （FISH-IF）的方法。
The dialdehyde glyoxal is an alternative chemical fixative that cross-links tissues faster than formaldehyde, retains higher antigenicity, and is less hazardous than either formaldehyde or glutaraldehyde. Here we present a glyoxal-based fixation protocol for use with Drosophila embryos. We describe steps to prepare acid-free glyoxal, fix embryos, and then stain with antibodies for immunofluorescence (IF). We also describe methods for RNA fluorescence in situ hybridization (FISH) and FISH plus IF (FISH-IF) using glyoxal-fixed embryos. This protocol was adapted for Drosophila embryos from the methods of Bussolati et al. and Richter et al. Glyoxal offers an alternative to formaldehyde for fixation of Drosophila embryos This protocol describes steps to prepare acid-free glyoxal and fix embryos A method for immunofluorescence staining of glyoxal fixed embryos is detailed RNA in situ hybridization methods are also described Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The dialdehyde glyoxal is an alternative chemical fixative that cross-links tissues faster than formaldehyde, retains higher antigenicity, and is less hazardous than either formaldehyde or glutaraldehyde. Here we present a glyoxal-based fixation protocol for use with Drosophila embryos. We describe steps to prepare acid-free glyoxal, fix embryos, and then stain with antibodies for immunofluorescence (IF). We also describe methods for RNA fluorescence in situ hybridization (FISH) and FISH plus IF (FISH-IF) using glyoxal-fixed embryos.
登录
查看更多内容
影响因子：
3.8
作者：
Rand, Matthew D.;Kearney, Alison L.;Dao, Julie;Clason, Todd
通讯作者：
Clason, Todd
影响因子：
64.5
作者：
NEUFELD, TP;RUBIN, GM
通讯作者：
RUBIN, GM
影响因子：
3.3
作者：
FARES, H;PEIFER, M;PRINGLE, JR
通讯作者：
PRINGLE, JR
影响因子：
3.3
作者：
Sharp, DJ;Brown, HM;Scholey, JM
通讯作者：
Scholey, JM
影响因子：
4.6
作者：
Bonello, Teresa T.;Choi, Wangsun;Peifer, Mark
通讯作者：
Peifer, Mark