University of Virginia, Department of Biology, Charlottesville, VA 22903, USA.
University of Virginia, Department of Biology, Charlottesville, VA 22903, USA.
STAR Protoc. 2023 Mar 17;4(1):101956. doi: 10.1016/j.xpro.2022.101956. Epub 2023 Feb 9.
A major barrier to immunostaining Caenorhabditis elegans is the permeabilization of the worm's cuticle without distorting or damaging its body. We present here a gel-based immobilization protocol for fixed worms coupled with chemical and enzymatic permeabilization. The permeabilization is followed by antibody staining and fluorescent imaging. This protocol can be modified for different fixatives, permeabilizing reagents, or molecular readouts. Unlike previous immunostaining approaches, such as freeze cracking or dissection, this protocol enables immunostaining across the whole body of a well-preserved C. elegans.
免疫染色秀丽隐杆线虫的一个主要障碍是在不扭曲或损坏其身体的情况下对其角质层进行渗透。我们在此介绍了一种基于凝胶的固定线虫的方法，该方法与化学和酶渗透相结合。渗透后进行抗体染色和荧光成像。该方案可以针对不同的固定剂、渗透试剂或分子读出进行修改。与以前的免疫染色方法（如冷冻开裂或解剖）不同，该方案能够对保存良好的秀丽隐杆线虫的整个身体进行免疫染色。
