Super-resolution at the 
AMI Imaging Center

Zeiss Elyra 7 with SMLM and SIM super-resolution

 

The AMI super-resolution fluorescence microscope is capable of capturing incredible biological detail such as... . This is achieved with one of two super-resolution approaches.

Structured Illumination Microscopy

Structured Illumination Microscopy (SIM) employs frequency information to calculate a final image with two-times greater resolution than a standard fluorescence microscope. This results in an eight-fold improvement in all three dimensions. On top of being capable of imaging samples in 3-D, the sample can also be captured over time with live cell microscopy. The AMI super-resolution microscope has an incubator that allows samples to be imaged in a temperature- and CO₂-controlled environment. This makes the microscope perfect at studying live cell cultures in enhanced detail.

Single Molecule Localisation Microscopy

Single Molecule Localisation Microscopy (SMLM) over-comes the natural resolution limit by calculating the location of single, clearly separated signals produced by single molecules. To detect spatially separate fluorescence signals within biological samples which are complex and dense is where the magic of SMLM becomes apparent. Using a variety of methods (collectively called "single molecule localisation") detection of signal can be separated in time rather than in space. This leads to the "blinking" of molecules which can be captured across many thousands or hundreds-of-thousands of sequential images. Following reconstruction, samples can be seen with detail approaching macromolecular scales. A prerequisite for this is that the sample is fixed so that no spatial changes occur during this long acquisition. This makes SMLM most suitable at imaging thin samples that have been fixed. 

 

Instrument specifications:

  • Auto focus for time-lapse acquisitions

  • Contrast methods: phase contrast, differential interference contrast (DIC), fluorescence (optional TIRF)

  • Temperature- and CO₂-control

  • Objectives: 100× 1.46 NA oil-immersion with correction collar, 63× 1.46 NA oil-immersion TIRF, 63× 1.4 NA oil-immersion, 40× 1.2 NA water-immersion with correction collar, 25× 0.8 NA multi-immersion, 10× 0.5 NA, 10× 0.3 NA

  • Laser excitation: 405nm 50mW, 488nm 500mW, 561nm 500mW, 642nm 500mW.

  • Camera pco.edge sCMOS

  • Andor EM-CCD camera iXon Ultra 897

  • Automated stage control

 

Instrument capabilities:

  • Resolution

  • Speed

  • Depth

  • Field of view

Instrument uses:
Apotome

TIRF

SMLM

SIM