Matters of light and death: reducing phototoxicity in live-cell imaging

Session

Poster Session Three

Authors

James Manton (1)

Affiliations

1. MRC Laboratory of Molecular Biology

Keywords

fluorescence, live-cell, phototoxicity, photobleaching, phase contrast, dark field

Abstract text

Photobleaching and phototoxicity plague live-cell fluorescence microscopy experiments. Light sheet illumination can significantly reduce levels of photodamage from those of confocal or epifluorescence microscopy, but there are still many situations in which the sample dies prematurely. These are particularly prevalent when attempting to image fast, dynamic processes in 3D.

Here, I will present recent work on mitigating photodamage, either by improving the stability of fluorescent molecules or by replacing them with other contrast agents. In particular, I shall describe attempts to:

1. engineer superior fluorescent proteins using AlphaFold;
2. reduce photobleaching via triplet-state depletion;
3. use gold nanoparticles to provide molecular specificity in scattering microscopy modalities;
4. enhance phase contrast microscopy with nanodiamonds;
5. reduce photobleaching via sample cooling.