Estimating numbers of intracellular molecules through analysing fluctuations in photobleaching
Date Available
2019-07-30Type
datasetData Creator
Bakker, ElcoSwain, P
Publisher
University of Edinburgh. School of Biological SciencesMetadata
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Citation
Bakker, Elco; Swain, P. (2019). Estimating numbers of intracellular molecules through analysing fluctuations in photobleaching, [dataset]. University of Edinburgh. School of Biological Sciences. https://doi.org/10.7488/ds/2594.Description
The impact of fluorescence microscopy has been limited by the difficulties of expressing measurements of fluorescent proteins in numbers of molecules. Absolute numbers enable the integration of results from different laboratories, empower mathematical modelling, and are the bedrock for a quantitative, predictive biology. Here we propose an estimator to infer numbers of molecules from fluctuations in the photobleaching of proteins tagged with Green Fluorescent Protein. Performing experiments in budding yeast, we show that our estimates of numbers agree, within an order of magnitude, with published biochemical measurements, for all six proteins tested. The experiments we require are straightforward and use only a wide-field fluorescence microscope. As such, our approach has the potential to become standard for those practising quantitative fluorescence microscopy.The following licence files are associated with this item: