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High-Density Super-Resolution Microscopy With An Incoherent Light Source And A Conventional Epifluorescence Microscope Setup

By Kirti Prakash

Posted 01 May 2017
bioRxiv DOI: 10.1101/132571

We report that single-molecule superresolution microscopy can be achieved with a conventional epifluorescence microscope setup and a Mercury arc lamp. The configuration termed as Omnipresent Localisation Microscope (OLM), is an extension of Single Molecule Localisation Microscopy (SMLM) techniques and allows single molecules to be switched on and off (‘blinking’), detected and localised. The use of a short burst of deep blue excitation can be further used to reactivate the blinking, once the blinking process has slowed or stopped. A resolution of 90 nm is achieved on test specimens (mouse and amphibian meiotic chromosomes). Finally, for the first time, we demonstrate that STED and OLM can be performed on the same biological sample using a simple buffer. It is hoped that such a correlative imaging will provide a basis for a further enhanced resolution.

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