New Playing Field for Localization Microscopy
2014 R&D 100 Winner
Prior to the introduction of the ELYRA P.1 with 3D PALM by Carl Zeiss Microscopy LLC, commercially available 3-D localization techniques resulted in images where the resolution was not uniform throughout a small capture volume due to location dependent localization precision. The utilization of these techniques would not allow researchers to model biological structures in a fast and reliable manner over large capture volumes.
Photo-activated localization microscopy (PALM) powers the 3D PALM module and allows users of the ELYRA P.1 super-resolution microscopy system to generate biological images with uniform precision and resolution through class-leading capture volumes of 51 μm x 51 μm x 1.4 μm for fluorescence microscopy localization experiments. Instead of acquiring an image from all fluorescent molecules at once, as in traditional methods, PALM images a few fluorescent molecules per acquisition over many acquisitions to build up an image. Appropriate fluorophores and mathematical modeling precisely determine molecule locations in the volume, which can be extended along the axial axis by acquiring multiple planes. These large structures, which can be acquired more quickly because of the per plane capture volume, enable researchers to study thicker biological structures with high uniform resolution. These advances help ELYRA P.1 with 3D PALM yield a uniform achievable resolution of 20 nm in the x and y axes and 50 nm in the z axis. This represents a tenfold spatial resolution increase over conventional fluorescence imaging that is limited by diffraction.
Photo-activated localization microscopy
Carl Zeiss Microscopy LLC
|Carl Zeiss Microscopy LLC's ELYRA P.1 with 3D PALM development team. Back row (l-r): Thomas Schenderlein, Benedikt Leubbers, Thomas Kalkbrenner, Wolfgang Schwerdtfeger and Klaus Weisshart. Front row (l-r): Stephan Sust, Michael Hilbert, Saskia Pergande and Philipp Sprengholz.|
The ELYRA P.1 with 3D PALM Development Team from Carl Zeiss Microscopy LLC
Benedikt Leubbers, Principal Developer
Klaus Weisshart, Principal Developer