Detection mCherry cells¶
For rabies experiments, a separate step is needed to detect starter cells which are usually labelled by the nuclear mCherry native fluorescence. The first step is to detect mCherry cells, which is preprocessing and is described here. The second step, to assign which of these mCherry cells have enough rolonies to be considered rabies positive is already science and out of the scope of iss-preprocess (but see iss-analysis).
Prerequisite¶
Data must be acquired, projected (using project_and_average) and registered (using register). Registration can be tricky, if it seems to fail, see select_reg_fluorescent_tile_parameters.ipynb for playing with the ops faster.
Automatic detection¶
flowchart TD
start[Start] --> save_unmixing_coefficients
unmix_coef[calculate_unmixing_coefficient];
batch_est(((segment_mcherry)));
subgraph save_unmixing_coefficients
unmix_coef --> diag_ref([plot_unmixing_diagnostics]);
end
unmix_coef --> batch_est;
batch_est --> remove_all_duplicate_masks;
style batch_est fill:#E1BEE7,stroke:#424242
style remove_all_duplicate_masks stroke:#000000,fill:#E1BEE7
style unmix_coef stroke:#000000,fill:#E1BEE7
style diag_ref fill:#BBDEFB,stroke:#616161,color:#000000
style save_unmixing_coefficients fill:#AAAAAA, stroke:#424242