Hi everyone,
I wondered if it is possible to back-project 2D classes onto their original micrographs (or, better, denoised micrographs), as has been done using the “in silico reconstitution” approach described by the Costa lab (https://www.sciencedirect.com/science/article/pii/S0959440X21001652). This would be a fantastic tool to interpret original EM denisty for highly flexible/heterogenous samples.
Many thanks in advance,
Matthias
Hi @mvorlaender,
Would the goal of this be to see relationships between particle location in mics and their orientation via their assigned 2D-class?
Thanks,
Kye
Hi Kye,
I like to think about it in the same way how subtomogram averages can be placed back into a tomogram to annotate the original data with a high signal to noise average (but in 2D instead of 3D). The goal would be to get insights into very flexible or heterogenous samples. For example, in the original publication by the Costa lab, this was used to visualize the orientations of two DNA helicases that are linked by a flexible DNA segment - each helicase could be refined separately to high resolution, but the entire assembly could not be resolved by SPA. Another application would be a scenario where you have different species (complexes, confirmations…) in a dataset that bind to the same structure (let’s say, a vesicle) and ask if different species mix randomly or form homotypic clusters.
I hope that helped?
Thanks a lot, Matthias