I am in need of an optimized protocol for staining young adults with MH27 using freeze crack method and methanol fixation. Does anyone have a protocol that they know will work? If you know of a better method than than freeze crack and methanol fixation it would be ok too.
The MH27 antibody works on worms fixed for 20 minutes in 3% formaldehyde (in PBS). You can use freeze-crack to try and open the worms and then apply the formaldehyde solution on the slide. Or you can do rapid repeated freeze-thaws of worms in tubes of 3% formaldehyde - this tends to break open only a subset of the worms (but they don’t float away, as they may with the slides).
Thanks for the advice. Do you have a paper that references this method?