Visualizing cellular life: From single cell imaging to in vivo single-molecule biochemistry
Pr. Ulrike Endesfelder
Max Planck Institute
In recent years and with the help of advanced fluorescence microscopy techniques, immense progress has been made in characterizing and quantifying the behavior of single cells on the basis of molecular interactions and assemblies in the complex environment of live cultures. Importantly, single-molecule imaging enables the in vivo determination of the stoichiometry and molecular architecture of subcellular structures, yielding detailed, quantitative, spatiotemporally resolved molecular maps and unraveling dynamic heterogeneities and subpopulations on the subcellular level.
You should watch this webinar if you are interested in a beginners guide on how to vamp up your wide field fluorescence microscope and sample preparations to single-molecule sensitivity, including many tips and tricks from my groups work. I will explain tools such as quantitative dual-color PALM imaging using dual fluorescent protein labeling in living cells, details of sample preparations like easy but precise drift correction by red-shifted beads or tracking of dense, highly dynamic single-molecule data.
Further introductory reading:
Vojnovic, I., Winkelmeier, J., & Endesfelder, U. (2019). Visualizing the inner life of microbes: practices of multi-color single-molecule localization microscopy in microbiology. Biochemical Society Transactions, 47(4), 1041-1065.
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