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# Post-correlation cryo-CLEM toolbox | ||
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This repository contains utilities to help with the on-lamella post-correlation cryo-CLEM workflow developed in the Chlanda Lab at Heidelberg University. | ||
For more information, see [our paper](https://doi.org/10.1101/2020.02.27.966739). | ||
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## FIJI plugin | ||
The FIJI plugin helps with handling the Z-stack tile scan acquired on the Leica cryo-CLEM wide-field microscope. | ||
The files are usually large (>100 GB) and therefore cannot be opened with FIJI or other image processing software. | ||
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The StitchTileScan command runs a maximum projection on a tilescan and stitches it to one image. | ||
This is useful for targeted cryo-FIB milling, as the image (exported as a PNG file) can be overlaid with an EM map of the grid using MAPS (ThermoFisher). | ||
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The ROIExtractor command allows you to easily extract single Z-stacks from the previously stitched map. | ||
You just need the original tile scan, the ROIset file and the stitched image (both are created by the StitchTileScan command) and and output directory. | ||
The stitched image will be opened and overlaid with the positions of the individual tiles. | ||
Whenever you click into a tile, the corresponding Z-stack will be extracted from the original tile scan and written into the output directory as a .tif. | ||
These are subsequently used for deconvolution and correlation with the matlab script. | ||
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## Matlab script | ||
The matlab script performs 3D rigid registration of two Z-stacks, one pre milling (pre-LM map) and one post EM acquisition (post-LM map), | ||
calculating a transformation matrix which is applied to the post-LM map. |