Epifluorescence microscop

Figure Bl.18.9. Epifluorescence microscope the object is excited from the top and the fluorescent light is emitted in all directions, as indicated by the multitude of arrows in the object plane. The fluorescent light within the aperture of the objective gives rise to the image, showing that much of the fluorescent light is lost for imaging.

In additional tests, the suppressor activities of pectins in the intact host/pathogen-interaction were investigated by injecting genetically resistant plants with pectic substances prior to inoculation with the rust fungus. Infected leaves were harvested, cleared, and stained with Calcofluor one week after inoculation, and fungal growth was assessed under the UV-epifluorescence microscope. 

Fluorescence microscopes used in immunohistochemistry are so called epifluorescence microscopes (i.e., excitation of the fluorescence and observation are from above (epi) the specimen). 

Confocal laser scanning microscopy (CLSM) in conjunction with specific staining techniques is best suited to elucidate intracellular trafficking and localization. CLSM is a specific epifluorescence microscopical technique capable of optical cross-sectioning with a spatial resolution of 1 /urn and below [41, 42],... 

Figure 5 Epifluorescence microscopic images of BT 20 cells (A) Cells incubated with Rhodamine-PE labeled stearyl triphenylphosphonium liposomes (B) mitochondria in BT 20 cells stained with Mite Tracker Red. Abbreviation. PE, phosphatidylethano-lamine. Source Prom Ref. 30.

Figure 29. Fiuman osteoblast-like MG 63 cells in cultures on material surfaces modified with carbon nanoparticles. A fullerene Cgo layers deposited on carbon fibre-reinforced carbon composites (CFRC), B fullerene C o layers deposited on microscopic glass coverslips, C terpolymer of polytetrafluoroethylene, polyvinyldifluoride and polypropylene, mixed with 4% of single-wall carbon nanohorns, D the same terpolymer with high crystalline electric arc multi-wall nanotubes, E diamond layer with hierarchically organized micro- and nanostmcture deposited on a Si substrate, F nanocrystalline diamond layer on a Si substrate. Standard control cell culture substrates were represented by a PS culture dish (G) and microscopic glass coverslip (FI). Immunofluorescence staining on day 2 (A) or 3 (B-Fl) after seeding, Olympus epifluorescence microscope IX 50, digital camera DP 70, obj. 20x, bar 100 pm (A, C, D, G,H)or 200 pm (B, E, F) [16].

Epifluorescence microscope equipped with appropriate filters (e.g., Zeiss Axioplan epifluorescence microscope with rhodamine filters). 

The technique of fluorescence spectral measurements has become very sensitive over the past decade. In order to obtain more information on the surface monolayers, a new method based on fluorescence was developed. It consisted of placing the monolayer trough on the stage of an epifluorescence microscope, with doped low concentration of fluorescent lipid probe. Later, ordered solid-liquid coexistence at the water-air interface and on solid substrates were reported. The theory of domain shapes has been extensively described by this method. 

Sase, I., Miyata, H., Corrie, J. E., Craik, J. S., andKinosita, K. Jr. (1995). Realtime imaging of single fluorophores on moving actin with an epifluorescence microscope. Biophys. J. 

Epifluorescence microscope An epifluorescent microscope (below) using an objective lens to perform two tasks To focus light upon the specimen being observed and to collect light being emitted by that specimen, which is fluorescent. 

Epifluorescence Microscope. The diagram shows the working of an upright epifluorescent microscope. The excitation beam (black ray) from the arc lamp passes through the excitation filter and dichroic mirror and directed toward the specimen. The return beam of emitted fluorescence wavelength (red, green and black rays) is reflected of the dichroic filter, emission filter, ocular and goes to the detector (eye or camera). Courtesy of Prof. J. Paul Robinson, Ph.D., Director of Purdue University Cytometry Laboratory, Purdue University, West Lafayette, IN, USA. 

Fluorescence microscope equipped with appropriate filter sets for detection of one or more fluorochromes and image capture system. If cells that grow flat are to be used, a conventional epifluorescence microscope is adequate. For thicker cells, a microscope equipped with a confocal laser scanning system is an advantage. 

Epifluorescence microscope with a 75W Xenon arc lamp fitted to a highly stabilized power supply (see Notes 2 and 3). 

An epifluorescence microscope such as the BH2-RFCA (Olympus, UK) fitted with objectives designed and suitable for transmission of fluorescence and UV light (e.g., xlO DPlan Apo 10 UV, numerical aperture = 0.4) should be used. 

The glass plate forms a part of a flow chamber through which a reagent solution may be introduced to provide the conditions for ATP hydrolysis. Rotation of the actin filaments can be viewed in an epifluorescence microscope (excitation at 546 nm and emission at 560-620 nm) and images then recorded with a television... 

First, samples are filtered over a black membrane filter (e.g. polyester or polycarbonate) with an appropriate pore size (i.e. 0.4 rm for bacteria and 0.8-2 (xm for eukaryotic cells). These screen filters are used because of their low background fluorescence and high contrast, which facilitates validation using the epifluorescence microscope (see below) (Brailsford 1996). Secondly, the retained cells are fluorescently stained using one or more physiological or taxonomic probes (see Section Fluorescent Stains for SPC). 

Finally, to further analyse their properties, the retained spots are visually inspected using an epifluorescence microscope equipped with a computer-driven moving stage. To that end, the sample holder is transferred to the motorized stage in exactly the same orientation as in the ChemScan. Highlighting of a green spot in the secondary scan map directs the microscope to the respective position on the membrane filter, allowing rapid and accurate visual discrimination between labelled cells and fluorescent particles ( validation ). 

Imaging experiments were performed using an epifluorescence microscope (BX 60, Olympus Optical, Tokyo, Japan) and an ultrasensitive, cryogenically cooled CCD camera (LN/CCD, Princeton Instruments, Roper Scientific, Trenton, NJ). A standard... 

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