Seitenhistorie

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Batch processing approach:

• Close the software

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• Reopen in batch mode? or just reopen? BatchLTfret (lifetime fret)
• In batch, you can also use threshold (lower, to get above background, and higher, to avoid pileup (eg. counts above the 5% of repetition rate (of 30 MHz)
• → ROI and thresholding, get a good one. calculate all and save, goes through them in batch mode.

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• We can there also enter the IRF, and apply it to all the to-be-measured batch files! "IRF to all"
• Here, make sure that you perform an initial fit on an image with counts that roughly match the number of counts in the IRF measurement; and ideally a cell that has counts in the average range of the dataset. Then "fit" and next, "fit to all".
• Fit all starts with all parameters, and generates an overview table!
• It might happen, that starting from a different image, some other images will not fit. That could be because the IRF peak is at a hugely different count (orders of magnitude different) compared to the peak of the sample.

More things to do with students:

• Compare fret flim to fret APB!
• Compare different groups fits (started using a different image) for the same sample set.
• Compare over/under right amount of fitting parameters (see waves of residuals).
• Compare batch vs. single processing.
• Remember the localization "dimension". Where is each interactor? What changes in that, upon treatment?