TY - BOOK

T1 - Experimental single molecule fluorescence microscopy for investigation of embryonic stem cell priming and transcription regulation

AU - Hvid, Kasper Graves

PY - 2019

Y1 - 2019

N2 - Embryonic development is both fascinatingly complex yet highly robust in replicating morphology and expression to a precise timing. The underlying mechanisms and signaling pathways are diverse and many remain undiscovered, yet with advancements in fluorescence imaging technology, we are now able to observe nanoscale structures and follow individual molecules inside living embryonic stem cells. This has the potential to give us a highly detailed look at how sub-cellular mechanisms are involved in making cell-fate choices. In this thesis, I will describe how we have implemented single molecule fluorescence microscopy to track and measure how individual core pluripotency transcription factor binds to chromatin inside living cells. Next, we implemented stochastic blinking of individual fluorophores to surpass the diffraction barrier and image the actin cytoskeleton of naive stem cells at nanoscale resolution. Finally, I show how we are able to localize and quantify active sites of transcription by hybridizing fluorescent probes to single molecules of mRNA.

AB - Embryonic development is both fascinatingly complex yet highly robust in replicating morphology and expression to a precise timing. The underlying mechanisms and signaling pathways are diverse and many remain undiscovered, yet with advancements in fluorescence imaging technology, we are now able to observe nanoscale structures and follow individual molecules inside living embryonic stem cells. This has the potential to give us a highly detailed look at how sub-cellular mechanisms are involved in making cell-fate choices. In this thesis, I will describe how we have implemented single molecule fluorescence microscopy to track and measure how individual core pluripotency transcription factor binds to chromatin inside living cells. Next, we implemented stochastic blinking of individual fluorophores to surpass the diffraction barrier and image the actin cytoskeleton of naive stem cells at nanoscale resolution. Finally, I show how we are able to localize and quantify active sites of transcription by hybridizing fluorescent probes to single molecules of mRNA.

M3 - Ph.D. thesis

BT - Experimental single molecule fluorescence microscopy for investigation of embryonic stem cell priming and transcription regulation

PB - Niels Bohr Institute, Faculty of Science, University of Copenhagen

ER -