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Correlative microscopy for Biology: bridging the gap betweem fluorescence and electron microscopy

L. Engelbrecht

Fluorescent Microscopy Unit, Central Analytical Facilities, Stellenbosch University

Email: lizeb@sun.ac.za

BY MSSA TECH FORUM

The quest to resolve cellular detail in the biological field has developed along two different avenues, namely light and electron microscopy. The first allows samples to be imaged with relatively little interference, making live cell imaging possible. The development of fluorescence microscopy has also made possible the localisation of specific molecules in the cellular environment. Electron microscopy achieves resolution much better than any light microscope, including the recently developed super-resolution microscopes. However, due to the requirements of sample preparation and imaging conditions, live cell imaging is not possible and the localisation of bio-molecules limited. In the past decade several laboratories have developed techniques to bridge the gap between fluorescence and electron microscopy, a technique known today as correlative light and electron microscopy (CLEM). For the first time in South Africa, a system capable of CLEM has been commissioned at Stellenbosch University this year, with a shuttle-and-find mode installed on the Zeiss 780 ELYRA PS1 system as well as the new Zeiss Merlin FE SEM. The Zeiss 780 ELYRA PS1 system is designed to perform confocal microscopy, SR-SIM and PALM/STORM super-resolution microscopy. The Zeiss Merlin FE SEM has an array of detectors of which the scanning transmission electron microscopy (STEM) detector, backscatter detectors and in-lens and out-of-lens secondary electron detectors would be of most importance for CLEM on biological samples. Here will be presented the capabilities of the system, the sample preparation procedures to follow and the first correlative images achieved on the system on a number of different biological samples.

 November 06, 2015
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