One experiment I’d LOVE to see (which I haven’t done yet because I’ve been working on the fixation stuff but might in the future) is to take YPF mice (sparse fluorescent labeling of entire neurons) and freeze / cryoprotect them in various combinations with various post-mortem delays, then do expansion microscopy and 3D imaging to see what happens to the YFP pattern. The idea is that you’d have the ground truth of what the intact neuron was and could see if it’s still traceable after the preservation attempt. I would be very impressed and ultimately convinced that a protocol maintains traceability if you could trace images with the YFP signal stripped and then have that tracing result match the YFP ground truth.
Certainly there’s some issues actually doing the experiments technically (which is one of the reasons I haven’t done them yet). It’s harder to do well than it might seem. But I do think it can be done, and I also think it’s never been easier because of expansion microscopy making it much easier and cheaper than it used to be by a factor of 100-1,000x.
One experiment I’d LOVE to see (which I haven’t done yet because I’ve been working on the fixation stuff but might in the future) is to take YPF mice (sparse fluorescent labeling of entire neurons) and freeze / cryoprotect them in various combinations with various post-mortem delays, then do expansion microscopy and 3D imaging to see what happens to the YFP pattern. The idea is that you’d have the ground truth of what the intact neuron was and could see if it’s still traceable after the preservation attempt. I would be very impressed and ultimately convinced that a protocol maintains traceability if you could trace images with the YFP signal stripped and then have that tracing result match the YFP ground truth.
Certainly there’s some issues actually doing the experiments technically (which is one of the reasons I haven’t done them yet). It’s harder to do well than it might seem. But I do think it can be done, and I also think it’s never been easier because of expansion microscopy making it much easier and cheaper than it used to be by a factor of 100-1,000x.