Abstract: Fluorescence microscopy has long been a powerful tool for studies in cellular and developmental biology, but suffers from a number of drawbacks. Chief among these are photobleaching and phototoxicity, which limit our ability to see what's going on while maintaining the sample in a state compatible with life. In this talk, I will describe our recent work on maximising the ratio of information gained to damage caused, looking at illumination delivery, emission handling, sample preparation and data processing.
In particular, I will describe our implementation of sub-cellular oblique plane microscopy, which enables gentle 3D imaging over hundreds of timepoints through optimised illumination. I will then introduce our projection imaging technique, which can increase acquisition rates up to 100-fold, and our multispectral imaging system, capable of imaging eight fluorescence markers with no penalty in spatiotemporal resolution. Examples of how this can be used to study the dynamics of organelles within living cells and endosomal trafficking will be shown. This will continue into a discussion of a new hands-free deconvolution algorithm, and conclude with a summary of current work-in-progress on improving the stability of fluorescent markers.

Biography: James is a Senior Investigator Scientist and Royal Society University Research Fellow at the MRC Laboratory of Molecular Biology, where he works to develop new technologies for biological microscopy. Trained as a physicist, his main current interests are in live-cell imaging and fluorescence photophysics.