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Department of Biosciences

Super-Resolution Microscopy

In optical microscopy, the resolution limit is defined by Abbe's principles. Generally this results in a lateral resolution (X-Y) limit of approximately 250nm and axial (Z) resolution of approximately 500nm.

Various optical techniques and sample preparation techniques result in this diffraction limit being broken. Durham's Biosciences Bioimaging facility are able to provide several super-resolution techniques which can be viewed below.

 

Super-Resolution Methods

Zeiss Airyscan

~ 140nm lateral resolution
Confocal microscope

3D-SIM

Structured Illumination Microscopy ~100nm lateral resolution
GE Healthcare OMX Version4

Localisation Microscopy

~20-50nm lateral resolution
GE Healthcare OMX Version4

Expansion Microscopy

Super-resolution via physical expansion
Diagram of expansion microscopy

Expansion Microscopy

Expansion microscopy (ExM), developed by Boyden and colleagues in 2015 ,is a microscopy methodology capable of producing super-resolution images on conventional microscopy equipment, through a combination of both physical and optical magnifications.

Further-more, the technique acts as a clearing method, allowing super-resolution microscopy deep within complex tissues, by removing the light-absorbing and light-scattering components, leaving only a three-dimensional fingerprint of labelled structures within the optically inert gel.

Expansion microscopy has developed over the years, initially expansion factors of ~4x have now been superseded by  20 fold expansion microscopy with many other iterations in between.

Here in Durham Bioimaging facility, we were the first to develop expansion microscopy in plant cells, read more here Expansion Microscopy of Plant Cells (PlantExM).