Super-resolution microscopy in Leicester


Technology feature: Super-resolve me: from micro to nano. Nature 526 (2015) 459–462.
What is super-resolution microscopy
?
Some review articles.

We can do SRRF super resolution reconstruction using microscope systems in the AIF but at the moment we don't have super resolution microscopes in the facility. There are several initiatives within the College to build super-resolution systems and these systems are already in use or are approaching test phase.

Dr Dmitry Cherny (Dept of Molecular and Cell Biology) is in charge of a home built TIRF microscope.

The system is built around inverted Axiovert 135 (Zeiss) microscope and equipped with water immersion 63x Zeiss objective and has 4 lasers: 488 nm, 532 nm, 561 nm and 643 nm. An illumination area is about 70x70 microns. The system is equipped with a single Andor iXon camera (512x512) and a beam splitter allowing detection of the signals from 2 or 3 fluorophores in one experiment. The system allows controlling polarization of the excitation light and detecting emissions in two orthogonal polarizations simultaneously. Stoichiometry of various complexes can be measured at the level of single molecules. Dmitry is open to discussions and collaboration using this microscope (dc114 or ext 7104).

Dr Andrey Revyakin (Dept of Molecular and Cell Biology) has built a STORM/PALM TIRF system named LESTASCOPE and is in the process of testing his system.

His inverted system is equipped with a 60x /1.49 NA Olympus objective and has 4 laser lines: 488 nm, 532 nm, 628 nm and 730 nm. A 405 nm laser will be added later this year. The Illumination area is 100x100 microns. Emission bands are Cy2 (510/30), Cy3 (580/60), Cy5 (675/45), and Cy7 (770/50) but this can be changed. Attached to the system are 4 Andor iXon ULTRA 1024x1024 water cooled camera, one for each channel for simultaneous multicolour imaging.  The system has drift correction (2 nm stability in XY and 5 nm in Z) and objective and sample temperature regulation. It is also equipped with a peristaltic pump for perfusion during real-time imaging. Ideally a 24 x 40 mm no1 coverglass is used for the sample but customized sample holders can be made/used. Andrey is open to discussion and collaboration using his microscope (both in TIRF mode or non-TIRF mode). Please contact Andrey (ar371 or 0772 921 8315) directly.

Prof Nick Hartell (Dept. of Neuroscience, Psychology and Behaviour) and Dr Andrew Hudson (Dept. of Chemistry) received funding via the BBSRC Alert14 initiative to build a functional super-resolution facility which uses novel technology developed at Leicester for fast, super-resolution multiphoton imaging in living systems. One microscope is built on an upright frame and will be equipped for concurrent electrophysiological recordings. A second microscope, built on an inverted frame, is designed for work with single cells or molecules. It will also allow d-STORM which will achieve a resolution of around 20 nm in fixed tissue. These systems are currently being built in room MSB 389 and should be operational in early 2016. To discuss potential projects please contact Nick (nh88, ext 3301) or Andrew (ah242, ext 2099) directly.

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