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> As the probes twinkle on and off, computational models estimate exactly where each molecule is located — and reconstruct a high-resolution image of the sample
by abeppu 1y ago
> As the probes twinkle on and off, computational models estimate exactly where each molecule is located — and reconstruct a high-resolution image of the sample.
How do time and motion fit in with these techniques? I'm dimly aware that the molecular machinery inside cells moves pretty fast, and that a lot of things move around randomly. In normal size ranges that kind of thing would naturally make it hard to get a clear picture. Do these imaging techniques require that stuff be frozen or specially prepared? Or do the techniques themselves work so fast that they can get a snapshot regardless?
- vonzepp 1y agoYes. This isn't for dynamic events.
- EColi 1y agoThe cells are "fixed" (with paraformaldehyde (PFA) for example), so yes these are snapshots but not because the technique is fast. These techniques can actually be quite slow because you need to collect enough blinks to reconstruct the final image.
- jkh1 1y agoYou can track things in live cells with MINFLUX, one of the recent super-resolution techniques coming from Stefan Hell's lab. Edit: add MINFLUX review: https://arxiv.org/pdf/2410.15902 https://arxiv.org/pdf/2410.15902
- ClaraForm 1y agoOther commenters are wrong. Live-cell can be done with older single-molecule localization microscopy using techniques like PAINT. The fluorophore is usually strategically added in a way that binding-unbinding events cause excitation. Algorithms can then infer identity of single fluorophores based on their excitation pattern/strength and can predict whether it's two distinct fluorophore molecules or the same molecule moving over multiple frames of image acquisition.