An Airyscan 2 confocal laser scanning microscope for the Wolfson Light Microscopy Facility
Lead Research Organisation:
University of Sheffield
Department Name: School of Biosciences
Abstract
Human diseases arise because of defects in fundamental biological processes. These processes depend on interactions between molecules within cells, between cells within tissues and tissues in the whole organism. In order to understand and identify effective therapeutics we need to understand the dynamic nature of biological processes and observing such processes in living cells is fundamental to this understanding. Visualisation both validates other experimental approaches but, importantly, often allows us to formulate new hypotheses. At the University of Sheffield biomedical researchers are exploring the fundamental way in which molecules, cells and tissues behave in order to understand what goes wrong in diseases such as cancer, infection, inflammation, deafness and neurodegeneration.
In order to observe biological processes we often attach fluorescent labels to components of interest and follow their behaviour using fluorescence microscopes. The time-scale of biological processes is highly variable: some processes happen very quickly, meaning that images need to be acquired speedily while others occur over a much longer time-scale of minutes to hours. There are a number of challenges associated with capturing fluorescence images because the exposure of cells to light can cause phototoxicity which damages cells and causes aberrant behaviour. Additionally, some fluorescent labels can be quite dim which makes the fluorescent signal difficult to detect.
The aim of this proposal is to obtain funding for the latest Airyscan 2 confocal laser scanning microscope (CLSM). This microscope will not only replace the functionality of two existing but very old fluorescent microscopes which are difficult to maintain, but will also improve our ability to observe very small structures within cells and/or to observe highly dynamic processes which occur very rapidly. The advanced performance of the Airyscan 2 CLSM will overcome current limitations in understanding fundamental mechanisms that underpin disease.
In order to observe biological processes we often attach fluorescent labels to components of interest and follow their behaviour using fluorescence microscopes. The time-scale of biological processes is highly variable: some processes happen very quickly, meaning that images need to be acquired speedily while others occur over a much longer time-scale of minutes to hours. There are a number of challenges associated with capturing fluorescence images because the exposure of cells to light can cause phototoxicity which damages cells and causes aberrant behaviour. Additionally, some fluorescent labels can be quite dim which makes the fluorescent signal difficult to detect.
The aim of this proposal is to obtain funding for the latest Airyscan 2 confocal laser scanning microscope (CLSM). This microscope will not only replace the functionality of two existing but very old fluorescent microscopes which are difficult to maintain, but will also improve our ability to observe very small structures within cells and/or to observe highly dynamic processes which occur very rapidly. The advanced performance of the Airyscan 2 CLSM will overcome current limitations in understanding fundamental mechanisms that underpin disease.
Technical Summary
The Wolfson Light Microscopy Facility (LMF) underpins biomedical research at the University of Sheffield. The LMF is a multiuser facility where researchers explore underpinning mechanisms of disease, focussing on molecular cell biology, including planar cell polarity, membrane trafficking, cell cycle, and cell fate; regenerative medicine in particular the use of human embryonic stem cells; infection and immunity including animal models of disease; and neurodegeneration using iPS cells. The LMF houses a suite of microscopes and our aim is to have appropriate microscope platforms for individual applications. Currently, standard confocal microscopy is provided by two heavily used and ageing microscopes dating from 2005 and 2012 and they are vulnerable to irreversible breakdown. The aim of this application is to obtain funding for an Airyscan 2 laser scanning confocal microscope. In addition to ensuring routine microscopy provision for the future, this microscope will provide enhanced imaging capability in terms of speed, sensitivity, and low phototoxicity. These advantages, coupled with the ability to capture wide fields-of-view, mean that this is a very powerful instrument which will have a wide range of uses for biomedical research within the LMF.
Organisations
Publications
Ganguli S
(2023)
Oncogenic Ras deregulates cell-substrate interactions during mitotic rounding and respreading to alter cell division orientation.
in Current biology : CB
Maib H
(2024)
Recombinant biosensors for multiplex and super-resolution imaging of phosphoinositides.
in The Journal of cell biology
Sheard TMD
(2023)
Differential labelling of human sub-cellular compartments with fluorescent dye esters and expansion microscopy.
in Nanoscale