The Integration of High Throughput Technologies for Drug Discovery

Sabbatini, Gregory P.; Shirley, William A.; Coffen, David L.

doi:10.1177/108705710100600402

Cited by 6 publications

(1 citation statement)

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“…High throughput or high content screening using fluorescence techniques requires simultaneous illumination of multiple samples, e.g., for drug discovery [31] or for examination of diseases accompanied by altered membrane properties, e.g., Morbus Alzheimer [32], Morbus Crohn [33] or Niemann-Pick disease [34]. For these purposes, we developed a microtiter plate reader system based on multiple TIR excitation and fluorescence detection [35].…”

Section: Tir-readermentioning

confidence: 99%

Total Internal Reflection Fluorescence Microscopy (TIRFM) – novel techniques and applications

Richter

Wagner

Schneckenburger

2020

MRAJ

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Total Internal Reflection Fluorescence Microscopy (TIRFM) has been established almost 40 years ago for studies of plasma membranes or membrane proximal sites of living cells. The method is based on light incidence at an angle above the critical angle of total internal reflection and generation of an evanescent electromagnetic field penetrating about 100 nm into a sample and permitting selective excitation of membrane proximal fluorophores. Two techniques are presented here: prism-type TIRFM and objective-type TIRFM with high aperture microscope objective lenses. Furthermore, numerous applications are summarized, e.g. measurement of focal adhesions, cell-substrate topology, endocytosis or exocytosis of vesicles as well as single molecule detection within thin layers. Finally, highly innovative combinations of TIRFM with Förster Resonance Energy Transfer (FRET) measurements as well as with Structured Illumination Microscopy (SIM) and fluorescence reader technologies are presented.

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