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- W2912503908 abstract "The development of PALM has allowed studies of single molecules with fluorescence below the diffraction limit both in fixed and live cells. The ability to localize single molecules with high resolution allows greater insight into their function, by analyzing their dynamics of movement in live cells such as diffusion, and structure and distribution by spatially quantifying the molecules. The question becomes what is the optimal imaging conditions that will lead to the best imaging results. This study sought to answer that by varying the excitation power and the pH of the imaging medium while imaging yeast (S. cerevisiae) with mEos2 tagged to the PH domain. By imaging our cells over a range of different excitation powers we show that in terms of trace length that there is an optimum power where single molecules can be tracked for the longest time on average, with the longer traces giving more accurate calculation of a diffusion coefficient. It has been shown for bulk studies of mEos2 in vitro that net signal intensity decreases with a decrease in pH. Our results show that the signal intensity of mEos2 is independent of the medium pH on the single molecule level and hence the localizations precision. By counting the number of molecules in fixed cells we find that fewer molecules fluoresce at low pH where we believe protonation be a factor in determining the ability to fluoresce." @default.
- W2912503908 created "2019-02-21" @default.
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- W2912503908 date "2019-02-01" @default.
- W2912503908 modified "2023-09-27" @default.
- W2912503908 title "Optimization of Single Molecule Palm Imaging Conditions using Meos2" @default.
- W2912503908 doi "https://doi.org/10.1016/j.bpj.2018.11.2363" @default.
- W2912503908 hasPublicationYear "2019" @default.
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