Super Resolution Fluorescence Microscopy

Eric Betzig, Stefan W. Hell and William E. Moerner were jointly awarded 2014 Nobel Prize in Chemistry for the development of super- resolved fluorescence microscopy. Microscope has been used as a powerful scientific instrument in the study of cell, cell biology and cellular functions. Cellular structure and objects occurs in the size range of tens to few hundreds nano meters however conventional light microscopy is only capable to handle cellular structure that are 200 to 350 nm apart (Schermelleh et al., 2010). Super resolution microscopy resolves more cellular structure at the macromolecular level.  Electron microscopy can resolve molecular and atomic structures with smaller wavelengths of electronic beam however energies of electrons have been found to be destructive for biological samples (Aguet, 3009)

Point spread function (PSF) can be defined as “the fixed size of the spread of a single point of light that is diffracted through a microscope” (Galbraith and Galbraith, 2011). Higher resolution microscopy helps to study smaller cellular structures than the PSF size of conventional microscopes. Cellular objects which are closer than PSF width of microscope appears as a single objects. Galbraith and Galbraith (2011) defined super-resolution microscopy as a technique which has at least double PSF width value than conventional microscopy.

Fluorophores are the molecules with an ability to fluoresce and fluorescence is the phenomenon where fluorophore emits light (Aguet, 2009). Fluoroscence microscopy has been used to distinguish cellular structure and specific protein by marking them (Aguet, 2009). Super resolution fluorescence microscopy has been used to study structures and functions of sub-cellular components and dynamic processes within the cell.

References

Aguet, F. (2009) Super-Resolution Fluoroscence Microscopy Based on Physical Models [Online] Available at www.bigwww.epfl.ch [Accessed on 21 November 2014].

Galbraith, C.G. and Galbraith, J.A. (2011) Super-Resolution Microscopy at a Glance. Journal of Cell Science, Vol 124, pp 1607-1611.

Schermelleh, L., Heintzmann, R. and Leonhardt, H.  (2010) A Guide to Super-Resolution Fluoroscence Microscopy. JCB Review, Vol 190(2), pp 165-175.

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