Application of Correction Curve
Hello, I was reading the paper "Single molecule mass photometry of nucleic acids" (doi: 10.1093/nar/gkaa632), and they eventually ended up applying a correction curve to renormalize their data to account for diffusion of nucleic acids. I wanted to ask if anyone had done something similar and if so, how they had done it. Did they use some external program to process the data or is this a feature in DiscoverMP? I attached a screenshot of the correction they used.
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In the manuscript you cite, flow chambers are used for sample introduction and measurement. Instead, Refeyn recommends using sample well cassettes and mixing your samples with the focusing buffer prior to measurement to minimize any diffusion considerations/artifacts. Consistency in the timing of sample addition, mixing, and starting the recording will also produce more reproducible counts and measurements.
However, if you want to evaluate whether diffusion is playing a role, you can use the temporal masks available in DiscoverMP to evaluate relative abundances of small mass vs large mass species during different stages of the recording. Alternatively, you can export the events files of the recording/s of interest, where the mass and frame (i.e. time point) of each particle counted can be found. Using this information you can run the integrations indicated in that manuscript to calculate any correction factors, but this is not a feature that could be run within DiscoverMP.