lbfcs2's Introduction

lbFCS2

This python package lbFCS2 allows to determine absolute docking strand numbers and local hyridization rates in individual DNA-PAINT clusters requiring only a single DNA-PAINT image aquistion, as published in 'Calibration-free counting of low molecular copy numbers in single DNA-PAINT localization clusters'. It is a revised framework of the lbFCS package, previously published in 'Towards absolute molecular numbers in DNA-PAINT'.

It is suited for the application to well-separated DNA-PAINT localization clusters containing only low molecular numbers of up to six docking strands.

Pre-requirements

For ...

... automated localization of the raw images,
... automated undrifting,
... and automated detection and isolation of all localization clusters within a DNA-PAINT image ...

it requires the picasso_addon based on the picasso package. A fixed release doi.org/10.5281/zenodo.4792396 of picasso_addon was already used in 'Tracking single particles for hours via continuous DNA-mediated fluorophore exchange' (corresponding python package can be found at SPT).

The complete installation instructions of the picasso_addon and picasso package and creation of the necessary conda environment compatible with picasso_addon, picasso, SPT and lbFCS2 can be found here picasso_addon: Installation instructions. The installation was tested on Windows 10.

All scripts for simple evaluation of data can be found in /scripts.

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