Gentle super-resolution imaging for any microscope

Session

Poster Session

Authors

Alexandra Schroeder (1), Caroline Peron Cane (1), Tom Sperber (1), Gabriel Sirat (1)

Affiliations

1. Telight

Keywords

Super-resolution, microscopy, life sciences, biology, gentle imaging, fluorescence, live-cell imaging, add-on system, phototoxicity, photobleaching

Abstract text

The ability to visualize the dynamics and structure of living samples has marked a revolutionary achievement of light microscopy in the life sciences.  It has paved the way for major breakthroughs in biomedical research as researchers can image and track interactions of subcellular structures within and around cells. Fundamentally, the versatility and relative simplicity of confocal laser-scanning microscopy has allowed it to enjoy its continued status as a major workhorse tool within biology. Unfortunately, in order to visualize finer details within cells and in the context of light-sensitive samples, the diffraction limit and phototoxicity are persisting challenges with confocal microscopy.

To address the first challenge of seeing beyond the diffraction limit (200-250 nm), many super-resolution microscopy solutions have successfully circumnavigated this and can visualize structures down to just a few nanometers in scale.  However, many of these techniques invite multiple drawbacks including fluorophore restrictions, long integration times and, in particular, higher levels of photo-damage. There remains a great need for simple, versatile, and gentle super-resolution imaging methods. 

Here, we present how super-resolution can be achieved without intense inhibition lasers or through photophysical manipulation of the sample. Using conical diffraction, this can be done quickly, passively, and achromatically for any visible wavelength of light, making it possible to produce structured illumination light patterns inherent to a scanned beam of light [1,2]. Such patterns can have features sharper than the diffraction limit, thereby directly optimizing the point-spread function of the system [1]. We present how conical diffraction is used as a beam-shaping tool that when integrated as an add-on to a microscope, can serve as a highly-sensitive, gentle imaging system with little-to-no photobleaching or phototoxicity on any sample [2]. 

References

1. G. Sirat, US Patent US9250185 - Method and device for superresolution optical measurement using singular optics. (Bioaxial SAS, 2010-2017).

2. J. Caron et al., "Conical diffraction illumination opens the way for low phototoxicity super-resolution imaging." Cell Adhesion & Migration. Vol. 8 No.5. 2014. DOI: 10.4161/cam.29358.