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#smlm

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Super-cool work from our clever colleagues at Cambridge -- whole-cell #3D #singlemolecule #lightfield imaging! #SMLM

@samofmidworld

High-density volumetric super-resolution microscopy
bioRxiv 2023.05.02.539032;
doi: doi.org/10.1101/2023.05.02.539

bioRxivHigh-density volumetric super-resolution microscopyVolumetric super-resolution microscopy typically encodes the 3D position of single-molecule fluorescence into a 2D image by changing the shape of the point spread function (PSF) as a function of depth. However, the resulting large and complex PSF spatial footprints reduce temporal resolution by requiring lower labelling densities to avoid overlapping fluorescent signals. We quantitatively compare the density dependence of single-molecule light field microscopy (SMLFM) to other 3D PSFs (astigmatism, double helix and tetrapod) showing that SMFLM enables an order-of-magnitude speed improvement compared to the double helix PSF by resolving overlapping emitters through parallax. We then experimentally demonstrate the high accuracy (>99.2 ± 0.1%, 0.1 locs μm−2) and sensitivity (>86.6 ± 0.9%, 0.1 locs μm−2) of SMLFM at point detection through whole-cell (scan-free) imaging and tracking of single membrane proteins in live primary B cells. We also exemplify high density volumetric imaging (0.15 locs μm−2) in dense cytosolic tubulin datasets. ### Competing Interest Statement CAIRN research has a co-development agreement with SFL and KOH at the University of Cambridge.

#introduction #superresolution #lightsheet #nanoscopy #smlm

Hi everyone!

My name is Ulrike, & I'm a #Physicist, Optical Scientist & #DataScientist passionate about community building/engagement, outreach, & teaching!

Currently, I'm working as an Optical Scientist & Project Team Lead at the #CarlZeiss AG (#teamzeiss, #zeiss) in #Oberkochen. Here, I'm part of the Corporate Research & Technology unit working on the latest optical trends for #imaging, #microscopy, & #metrology.

Let's connect!

📢 📢New lab preprint!!! Check out @louvel_vincent 's work, he pushed expansion microscopy to SMLM-like resolution by optimizing U-ExM and interative expansion. Check out this thread to learn more👇
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RT @louvel_vincent
Thrilled to present my PhD work about the development of #iUExM 🥳
Here, we introduce an optimized versatile protocol using iterative expansion to reach #SMLM resolution on conventional widefield or confocal fluorescen…
twitter.com/louvel_vincent/sta