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Topic: Total internal reflection fluorescence microscope

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Microscope

	Total internal reflection fluorescence microscope - Wikipedia, the free encyclopedia
		A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nm, can be observed.
		When these molecules are excited and detected with a conventional fluorescence microscope, the resulting fluorescence from those fluorophores bound to the surface is often overwhelmed by the background fluorescence due to the much larger population of non-bound molecules.
		Importantly, the fluorescence excitation energy of the evanescent wave is the same as the energy of the wavelength of the light that was totally internally reflected.
	en.wikipedia.org /wiki/Total_internal_reflection_fluorescence_microscope (343 words)

	Olympus Microscopy Resource Center: Specialized Microscopy Techniques - Total Internal Reflection Fluorescence ...
		Regardless of the basic microscope design, a majority of the currently utilized TIRFM configurations rely on an added prism to direct laser illumination toward the interface where total internal reflection occurs, which is in the specimen conjugate plane of the microscope.
		However, when the microscope stage is moved during focus, the illuminated region may move laterally as the specimen is focused, depending upon whether the focusing lens and beam director are mounted on the laboratory bench or the microscope stage.
		In general, total internal reflection is more difficult to implement on a microscope that has a moveable stage because focus adjustments require realignment of the laser beam into the focal plane and optical axis of the microscope.
	www.olympusmicro.com /primer/techniques/fluorescence/tirf/tirfconfiguration.html (6510 words)

	Microscope Encyclopedia Article @ NaturalResearch.org (Natural Research) (Site not responding. Last check: 2007-11-02)
		Van Leeuwenhoek's microscopes consisted of a single, small, convex lens mounted on a plate with a mechanism to hold the material to be examined (the sample or specimen).
		Compound optical microscopes can magnify an image up to 1000× and are used to study thin specimens as they have a very limited depth of field.
		Anton van Leeuwenhoek (1632-1723) is generally credited with bringing the microscope to the attention of biologists, even though simple magnifying lenses were already being produced in the 1500's, and the magnifying principle of water-filled glass bowls had been described by the Romans (Seneca).
	www.naturalresearch.org /encyclopedia/Microscope (1197 words)

	Nikon MicroscopyU: Fluorescence Microscopy - Total Internal Reflection Fluorescence
		Total internal reflection is only possible in situations in which the propagating light encounters a boundary to a medium of lower refractive index.
		The basic concept of total internal reflection fluorescence is schematically illustrated in Figure 1, in which specimen cells incorporating fluorescent molecules (green fluorophores in the figure) are supported on a glass microscope slide.
		Total internal reflection does not occur suddenly as a new phenomenon at the critical angle, but a continuous transition is followed from predominant refraction with a small amount of reflection, to total reflection when the critical angle is exceeded.
	www.microscopyu.com /articles/fluorescence/tirf/tirfintro.html (4510 words)

	Encyclopedia :: Microscope (Site not responding. Last check: 2007-11-02)
		The stereo microscope is often used to study the surfaces of solid specimens or to carry out close work such as sorting, dissection, microsurgery, watch-making, small circuit board manufacture or inspection, and the like.
		Compound microscope, an instrument consisting of a combination of lenses such that the image formed by the lens or set of lenses nearest the object (called the objective) is magnified by another lens called the ocular or eyepiece.
		Achromatic telescope, or microscope, one in which the chromatic aberration is corrected, usually by means of a compound or achromatic object glass, and which gives images free from extraneous color.
	www.hallencyclopedia.com /Microscope (2035 words)

	Nikon MicroscopyU: Fluorescence Microscopy
		Fluorescence Illumination in Stereomicroscopy - The application of stereomicroscopes for GFP observation is now so prevalent that stereo fluorescence illuminators are more frequently referred to as GFP illuminators, even though they can be utilized for many other applications in both the life sciences and the electronics manufacturing industry.
		The fluorescence illumination reveals which organisms are producing the fluorescent protein and the stereoscopic vision coupled to a large field of view and ample working distance enables observers to conduct experiments with forceps, pipettes, or micromanipulators.
		Typically, the fluorescence illuminator consists of a xenon or mercury arc lamp contained in an external lamphouse that is attached to the microscope via an intermediate tube (or vertical illuminator) positioned between the microscope zoom body and observation tubes.
	www.microscopyu.com /articles/fluorescence (1980 words)

	Fluorescence microscope - Wikipedia, the free encyclopedia
		A Fluorescence Microscope is a light microscope used to study properties of organic or inorganic substances using the phenomena of fluorescence and phosphorescence instead of, or in addition to, reflection and absorption.
		Most fluorescence microscopes in use are Epi-Fluorescence Microscopes (ie : excitation and observation of the fluorescence are from above (epi) the specimen).
		These microscopes have become an important part in the field of biology, opening the doors for more advanced microscope designs, such as Confocal laser scanning microscope and the Total internal reflection fluorescence microscope.
	en.wikipedia.org /wiki/Fluorescence_microscope (122 words)

	Olympus Microscopy Resource Center: Specialized Microscopy Techniques - Total Internal Reflection Fluorescence (TIRF)
		Total internal reflection fluorescence microscopy (TIRFM) is an elegant optical technique utilized to observe single molecule fluorescence at surfaces and interfaces.
		Excitation by an Evanescent Wave - Total internal reflection fluorescence is employed to investigate the interaction of molecules with surfaces, an area which is of fundamental importance to a wide spectrum of disciplines in cell and molecular biology.
		Trapezoidal Prism Microscope Configuration - The simplest approach to achieve total internal reflection from a culture chamber on an inverted microscope is direct laser illumination through a glass cube, prism, or trapezoidal block positioned on top of the chamber.
	www.olympusmicro.com /primer/techniques/fluorescence/tirf/tirfhome.html (1596 words)

	Olympus FluoView Resource Center: Total Internal Reflection Fluorescence Microscopy
		Total internal reflection microscopy (TIRFM) is an optical technique used to observe single molecule fluorescence.
		Total internal reflection is an optical phenomenon that can be employed to observe events occuring at boundaries.
		In fluorescence mode, having focused on the beads, the bead fluorescence is very difficult to distinguish because of the obscuring background fluorescence from the cheek cells.
	www.olympusfluoview.com /applications/tirfmintro.html (1285 words)

	Microscope - (Site not responding. Last check: 2007-11-02)
		Image:Igmicro.JPG Image:Compound microscope john cuff 1750.jpg A microscope (Greek: micron = small and scopos = aim) is an instrument for viewing objects that are too small to be seen by the naked or unaided eye.
		In most microscopes, the eyepiece is a compound lens, which is made of two lenses one near the front and one near the back of the eyepiece tube forming an air separated couplet.
		Optical microscopes are restricted in their ability to resolve features by a phenomenon called diffraction which, based on the numerical aperture (NA or $A_N$ ) of the optical system and the wavelengths of light used ( $\lambda$ ), sets a definite limit (d) to the optical resolution.
	psychcentral.com /psypsych/Microscope (1812 words)

	Single Molecule Imaging of Green Fluorescent Proteins in Living Cells: E-Cadherin Forms Oligomers on the Free Cell ...
		The argon ion laser beam (488 nm) was totally internally reflected at the coverslip-medium interface (the incident angle was set at 66°), and an evanescent field was formed (1/e penetration depth was ~100 nm).
		In B (between 0 and 70 AU of the fluorescence intensity) and C, the quadruple and double Gaussian functions, respectively, were used for fitting by assuming the width evaluated in D (for the peaks representing apparent dimer and greater oligomers, the width was multiplied by the number of monomers in an oligomer).
		that the fluorescence intensity for the basal peak of the quantized
	www.biophysj.org /cgi/content/full/80/6/2667 (7139 words)

	Biophotonics - Wikipedia, the free encyclopedia
		This refers to emission, detection, absorption, reflection, modification, and creation of radiation from living organisms and organic material.
		In microscopy, the development and refinement of the confocal microscope, the fluorescence microscope, and the total internal reflection fluorescence microscope all belong to the field of biophotonics.
		The specimens that are imaged with microscopic techniques can also be manipulated by optical tweezers and laser micro-scalpels, which are further applications in the field of biophotonics.
	en.wikipedia.org /wiki/Biophotonics (227 words)

	Spectral Fluctuation of a Single Fluorophore Conjugated to a Protein Molecule -- Wazawa et al. 78 (3): 1561 -- ...
		reflected at the incident angle of 69° to the norm at the silica
		The fluorescence emission from the samples collected by a microscope objective (MO) is switched for either imaging or spectroscopy by a removable mirror (RM).
		The total fluorescence intensity was obtained by integrating fluorescence intensity for wavelengths from 540 to 660 nm for each spectrum and plotting as a function of time.
	www.biophysj.org /cgi/content/full/78/3/1561 (4559 words)

	Fluorescence Microscope, and other Fluorescence Microscope information (Site not responding. Last check: 2007-11-02)
		Fluorescence is the most rapidly expanding microscopy technique in both the medical and biological sciences, a fact which has spurred the development of more sophisticated microscopes and fluorescence accessories.
		BX51 Upright Microscope - The modern upright epi-fluorescence microscope is equipped with a...
		Reflected light fluorescence microscopy is overwhelmingly the current method of choice for widefield investigations with non-coherent light sources, as well as those conducted with laser scanning confocal and multiphoton instruments.
	www.maxiscopes.co.uk /directory/fluorescence-microscope.html (458 words)

	Live cell imaging facility to aid research
		Four new microscopes comprising the Live Cell Imaging Core Facility allow researchers in both the clinical and the basic sciences to view living cells and track biological processes with three-dimensional clarity.
		The total internal reflection fluorescence microscope, or TIRF, is used primarily for studies of proteins and receptors in cell membranes.
		A deconvolution microscope, used especially for examining yeast and bacteria, and a laser-scanning confocal microscope, employed for larger and thicker specimens, also are available.
	www.utsouthwestern.edu /utsw/cda/dept146133/files/227177.html (330 words)

	Olympus FluoView Resource Center: Instrument Brochures
		microscopes and available support equipment for the innovative microscopist, the appropriate brochures may be downloaded in this section.
		The inverted instrument is also equipped with new fluorescence accessories and a V-shaped optical path for improved sensitivity in critical applications, while an external power supply ensures a stable platform for time-lapse observations.
		TIRFM Total Internal Reflection Fluorescence Microscope System - A technique especially useful in studies of cellular membrane function and single molecule events, total internal reflection microscopy is readily facilitated by the innovative Olympus TIRFM microscope system.
	www.olympusconfocal.com /brochures (1603 words)

	Insulin Stimulation of GLUT4 Exocytosis, but Not Its Inhibition of Endocytosis, Is Dependent on RabGAP AS160 -- ...
		Total HA-GLUT4-GFP patterns were determined by the GFP fluorescence, and HA-GLUT4-GFP on the surface was determined by indirect immunofluorescence of the HA epitope in fixed cells.
		Total HA-GLUT4-GFP patterns were determined by the GFP fluorescence, surface HA-GLUT4-GFP was determined by indirect immunofluorescence of the HA epitope in fixed cells, and expression of the FLAG-tagged AS160-4P was determined by indirect immunofluorescence after permeabilization of the fixed cells.
		The TIRF GFP fluorescence is normalized to the total amount of HA-GLUT4-GFP per cell measured in the epifluorescence/wide field mode of the microscope (WF).
	www.molbiolcell.org /cgi/content/full/15/10/4406 (6428 words)

	Imaging Constitutive Exocytosis with Total Internal Reflection Fluorescence Microscopy -- Schmoranzer et al. 149 (1): ...
		rise in the total, peak, and width of the fluorescence intensity.
		Total intensity, peak intensity, and square of the Gaussian width were plotted on the same time axis for carriers with long and short rise times (time is marked relative to the start of the rise phase).
		a slight increase in the total and peak intensities was evident
	www.jcb.org /cgi/content/full/149/1/23 (6070 words)

	Single-molecule imaging analysis of Ras activation in living cells -- Murakoshi et al. 101 (19): 7317 -- Proceedings of ... (Site not responding. Last check: 2007-11-02)
		The fluorescent spots shown in the BodipyTR channel appeared as a result of FRET from YFP on Ras to BodipyTR on GTP (arrows).
		The fluorescent spots in the figure may look aggregated in some areas, but their separations are rather clear in the original tape (see Movie 2).
		However, single fluorescent molecules are photobleached quickly under the conditions of single-molecule detection, causing insufficient averaging.
	www.pnas.org /cgi/content/full/101/19/7317 (4849 words)

	GMx33 Associates with the Trans-Golgi Matrix in a Dynamic Manner and Sorts within Tubules Exiting the Golgi -- Snyder ...
		The actual fluorescence intensity for each cell over time was converted to a percentage of the total recovery.
		Fluorescence values obtained immediately after bleaching were used as 0%.
		Therefore, the total fluorescence never recovered to the initial prebleach levels, and recovery level was inversely proportional to the time of bleaching.
	www.molbiolcell.org /cgi/content/full/17/1/511 (7904 words)

	Biophysical Journal: Single molecule imaging of green fluorescent proteins in living cells: E-cadherin forms oligomers ...
		ABSTRACT Single green fluorescent protein (GFP) molecules were successfully imaged for the first time in living cells.
		GFP linked to the cytoplasmic carboxyl terminus of E-cadherin (E-cad-GFP) was expressed in mouse fibroblast L cells, and observed using an objective-type total internal reflection fluorescence microscope.
		Based on the fluorescence intensity of individual fluorescent spots, the majority of E-cad-GFP molecules on the free cell surface were found to be oligomers of various sizes, many of them greater than dimers, suggesting that oligomerization of E-cadherin takes place before its assembly at cell-cell adhesion sites.
	www.findarticles.com /p/articles/mi_qa3938/is_200106/ai_n8958914 (423 words)

	DEPARTMENT-NAME: University of Leicester
		This instrument is used in studies using caged substrates and photophysics of green fluorescent protein variants.
		A custom-built total internal reflection fluorescence microscope, with 3 laser sources and various detection systems (dual ICCD cameras and avalanche photodiodes), is used for single molecule kinetic investigations.
		It comprises five high resolution fluorescence microscopes, two with microinjection, for analysis of both live and fixed cells.
	www.le.ac.uk /by/main_pages/general.html (1420 words)

	[No title] (Site not responding. Last check: 2007-11-02)
		TIRF (Total Internal Reflection Fluorescence) Microscope enables the visualization of a single molecule fluorescence in living cells.
		The evanescent wave resulting from total internal reflection(TIR) causes excitation of fluorescent molecules in an optical section, typically less than 100nm, without exciting molecules throughout the specimen.
		Evanerscent wave illumination provides such high signal to noise (S/N) that it is possible to observe a single molecule activity in living cells where they are in contact with the coverglass.
	www.techinst.com /applications_Systems.asp (336 words)

	Differential actin binding along the PEVK domain of skeletal muscle titin -- Nagy et al. 117 (24): 5781 -- Journal of ...
		The total filament length per field of view at the initial concentration of 25 mM KCl, were 115 µm, 145 µm and 195 µm for the PEVKI, -II and -III segments, respectively.
		The total filament length per field of view at the initial concentration of 25 mM KCl were 400 µm and 1140 µm for the PPAK and pure polyE fragments, respectively.
		The total polyE concentrations for the S-P lane pairs in this assay were: 0.036, 0.048, 0.060, 0.072, 0.084 and 0.096 mg/ml from left to right of gel.
	jcs.biologists.org /cgi/content/full/117/24/5781 (5048 words)

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