Single-Molecule Fluorescence Observed with Mercury Lamp Illumination

We demonstrate that it is possible to observe single fluorescent molecules using a standard fluorescence microscope with mercury lamp excitation and an inexpensive cooled charge-coupled device (CCD) camera. With this equipment, we have been able to observe single molecules of tetramethyl-rhodamine,...

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Veröffentlicht in:	BioTechniques 1999-11, Vol.27 (5), p.1008-1014
Hauptverfasser:	Unger, M, Kartalov, E, Chiu, C.-S, Lester, H.A, Quake, S.R
Format:	Artikel
Sprache:	eng
Schlagworte:	Biological and medical sciences Diverse techniques Fundamental and applied biological sciences. Psychology Green Fluorescent Proteins Histidine Image Processing, Computer-Assisted Light Luminescent Proteins - ultrastructure Mercury Microscopy, Fluorescence - instrumentation Microscopy, Fluorescence - methods Molecular and cellular biology Peptides - chemistry Plasmids - genetics Rhodamines - chemistry Streptavidin - ultrastructure
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Beschreibung
Zusammenfassung:	We demonstrate that it is possible to observe single fluorescent molecules using a standard fluorescence microscope with mercury lamp excitation and an inexpensive cooled charge-coupled device (CCD) camera. With this equipment, we have been able to observe single molecules of tetramethyl-rhodamine, rhodamine 6G, fluorescein isothiocyanate and green fluorescent protein. Immobilized molecules were observed both in air and in aqueous solution.
ISSN:	0736-6205 1940-9818
DOI:	10.2144/99275rr01