Autofocus-STED

Often, live cell measurements face the challenge that the dynamics of interest in the specimen take place on long time scales, sometimes hours up to days. 

  • An observation over a time of one hour at axial (z-) resolutions below 80 nm means that sample drift must be reduced to below 1.5 nm per minute.

A feat clearly not possible without active stabilization. That’s why Abberior Instruments offers a continuous autofocus module that locks the absolute z-position of the sample, while superresolution images can be simultaneously recorded without loss of performance. 

Experiments with short acquisition times benefit as well, because typical sample drift with a heated sample chamber can easily be in the range of tens of nanometers per minute, too, spoiling superresolution imaging even over the course of a few minutes. Also, for measuring in a less-than-optimally air-conditioned room, active stabilization with our autofocus module is the remedy.

Three different measurement modes have been implemented that can be fully controlled using our Imspector software: 

 

 

 

The autofocus module operates unobtrusively at wavelength ranges in the near infrared, well above any wavelength used with STED or fluorescent imaging. It is only available for Abberior Instruments STED and RESOLFT microscopes and can be upgraded anytime before or after installation of the microscope.

 

Measurement examples here.