However, it is difficult to study systems and phenomena at temperatures above 37 Â☌, because the index-matching fluids used with high-numerical-aperture objective lenses can conduct heat from the sample to the lens, and sustained exposure to high temperatures can cause the lens to fail. This has opened the door for many chemical and biological systems to be studied at native temperatures at the single-molecule level both in vitro and in vivo. Stavrakis, Stavros Quake, Stephen R.Īlthough single-molecule fluorescence spectroscopy was first demonstrated at near-absolute zero temperatures (1.8 K), the field has since advanced to include room-temperature observations, largely owing to the use of objective lenses with high numerical aperture, brighter fluorophores and more sensitive detectors. Colloidal lenses allow high-temperature single-molecule imaging and improve fluorophore photostability
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