Super Resolution Microscopy in Total Internal Reflection Fluorescence (SR-TIRF)

Abstract

One of the key tools of bio-medical research are light microscopes and to be more specific fluorescent microscopes. A fluorescent microscope allows the scientist to view tagged parts of a Cell, Virus, etc... at a molecular level so it's behaviour can be monitored.

Such fluorescent microscopes are driving modern bio-medical research to aid in our understanding of illness, disease and infection and in turn allow the development of improved treatments and possible cures.

However, the performance of these light microscopes has been hampered by the previous limit of optical resolution which was defined by a German Physicist in the 19th Century, Ernst Abbe, and as such carries his name, Abbe's Law.

Abbe's Law determined that the maximum achievable resolution of a light microscope is given by the wavelength of the light being used to view it (in many cases around 550nm) divided by twice the numerical aperture of the lens used for imaging (in modern microscopes the Maximum NA = 1.4).

This limited the resolution at which light microscope could observe biological systems and interactions to >200nm, as scientists developed an improved understanding of biology this was becoming a frustrating bottle neck for furthering research, until that is the development of super-resolved or Nanoscopy techniques and applications. This new emerging field was highlighted by the 2016 Nobel Price in Chemistry which was awarded for the development of super-resolved fluorescent microscopy.

However, whilst these new techniques did allow light microscope to resolve as low as 20nm the techniques were quite complex and could not always be used for every tool within the field of light microscopy which the scientist may have wished to use.

One such tool is Total Internal Reflection Fluorescence (TIRF) Microscopy. TIRF is currently used in Cell biology to view events which happen at the surface of cells, such events can play a major roll in the behaviour of the cell and scientists who research cancer (for example) are trying to understand the movement and spread of cancer cells by looking at events at the cellular surface using TIRF microscopes.

The instrument we propose to develop within this project will double the spatial resolution of a regular TIRF microscope enabling features <100nm to be imaged. This will offer the global science community an imaging system which can do true super-resolved live cell imaging with TIRF (SR-TIRF) allowing further advancements within bio-medical research.

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Participant

VISITECH INTERNATIONAL LIMITED

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