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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

GE Healthcare OMX V4.0

The system is an Applied Precision OMX BLAZE Super-Resolution microscope supplied by GE Healthcare.

The microscope can be utilised in several different modes;

  • Widefield (conventional) mode
  • 3D - SIM (Structured Illumination Microscopy)
  • TIRF (Total Internal Reflection) mode
  • Localisation microscopy - utilises images generated in TIRF mode

 

GE Healthcare OMX Version4

General Specification

Inverted microscope

Environmental control - temperature, humidity & CO2

Widefield (Conventional) Mode

The OMX can be used in its most simplest conventional mode to act as a very precise widefield microscope.

The fast camera speed of the OMX, dependent on camera exposure time, can run at approximately 40 fps

Ex 381-410nm/Em max. 436nm   DAPI

Ex 461-493nm/Em max. 528nm   FITC

Ex 562-581nm/Em max. 609nm   A568

Ex 638-581nm/Em max. 683nm    Cy5 

Structured Illumination Specification

3D-SIM (Structured illumination Microscopy) which provides an 8 fold improvement in volumetric resolution (xy 100nm by z 250nm)

Solid state laser lines

405nm

445nm

488nm

514nm

568nm

642nm

TIRF Specification

Total Internal Reflection (TIRF) microscopy

The OMX utilises a novel Ring TIRF module - this rotates the laser beam rapidly at the back focal plane of the objective lens creating a highly uniform TIRF illumination.

TIRF mode is utilised for DeltaVision Localization Microscopy - capable of lateral resolution approximately 20-50nm

Objective Lenses

  • 40 x oil - NA 1.3
  • 60 x oil - NA 1.42
  • 60 x silicone - NA 1.3

 

  • TIRF  60 x - NA 1.49
  • TIRF 100 x - NA 1.49