Light sheet fluorescence microscopy for high speed 3-D imaging: applications to the study of heart disease and cancer cell invasion

Lead Research Organisation: Imperial College London
Department Name: Physics

Abstract

Light sheet fluorescence microscopy enables high speed 3D fluorescence imaging of live cells with low photobleaching and phototoxicity. This project will develop novel light sheet microscopy technology to study live cells in high speed in 3D and to perform 3D screening of cells in multiwell plates. The first part of the project will develop a novel inverted light sheet fluorescence microscope system using adaptive optic elements for rapid refocusing for high speed 3D imaging as well as a second mode of operation for lower speed but high spatial resolution imaging. This will enable the structure of cardiomyocytes to be studied at high resolution and subsequently in 3D at high speed with accurate spatial registration of the two datasets. The existing oblique plane microscope setup will also be used during the project to study the invasion of cancer cells in 3D cultures and a significant part of the project will be to develop and accelerate the image acquisition, preprocessing and analysis pipeline.

Publications

10 25 50

Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/N509486/1 01/10/2016 31/03/2022
1858780 Studentship EP/N509486/1 01/10/2016 31/01/2021 Terence Wright
 
Description In this work, a deformable mirror (adaptive optic) has been used to refocus a microscope objective. This refocusing system has been applied to an inverted light sheet microscope. Deformable mirrors can change their profiles at speeds of the order of kHz and as a result can be used to refocus a microscope very rapidly - without needing to physically move the sample or the objective. The deformable mirror can be used to volumetrically scan an objective at video frame rates. Currently with the mirror scanning at a rate of 26 volumes/sec high resolution images can be obtained over a 200×200 mm2 field of view in the sample with a 80 mm defocus range. This should be sufficiently large to investigate high speed cellular processes.
Exploitation Route This form of deformable mirror based high speed volumetric scanning can in principal be applied to any situation which requires rapid refocusing such as imaging cardiomyocytes.
Sectors Pharmaceuticals and Medical Biotechnology