By Haney, Steven A
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I) Increase sensitivity or gain of microscope detector and reduce proportionally the time that the sample is exposed to the light source, or (ii) reduce or eliminate free radical production within the sample by means of antioxidants or free radical scavengers such as carotenoids [6, 7]. In fixed end-point assays, photobleaching is normally not a problem because each sample is acquired in a single acquisition scan. 5 LOCALIZATION OF FLUOROPHORES WITHIN CELLS As mentioned earlier, a vast number of fluorescent dyes, probes, and markers are currently available.
He observed that the light emitted from minerals such as fluorspar was of a longer wavelength than that of the incident light illuminating these minerals. The term Stokes’ shift was subsequently coined in his honor to describe this phenomena [4]. At the atomic level, this shift from wavelength to longer wavelengths occurs when electrons are energized or excited from their ground state by incident light rays. Eventually, these electrons drop back to their stable ground state and rerelease or emit their excess energy as light of a longer wavelength [5].
All of these will be discussed in this book. In each case, the core process of HCS will be described but linking it to the needs of the laboratory will depend on the HCS team. KEY POINTS 1. HCS represents the integration of diverse skills. In general, scientists working in HCS will have a high level of expertise in a few areas, but will rely on others with complementary expertise to form a robust team. 2. An appreciation of the power of HCS is invaluable, ironically because there are many occasions where a simple and efficient assay is optimal.