Fluorescence microscopy microscope

TIRFM total internal reflection fluorescence microscopy/ microscope 3.7c.iv... 

It is interesting to note the analogy of developments in light microscopy during the last few decades. The confocal microscope as a scaiming beam microscope exceeds by far the nomial fluorescence light microscope in resolution and detection level. Very recent advances in evanescent wave and interference microscopy seem to promise to provide even higher resolution (B1.18). 

Fluorescence microscopy measurements were performed with a Zeiss Axioplan microscope (Zeiss Co. Germany) equipped with a mercury lamp and a 40X objective. Images were acquired by CCD camera CH250 (Photometrix Co., Germany) cooled at -40°C by a liquid cooling unit CH260 (Photometrix Co., Germany). 

Tlie morphology of some bacteria, especially those that form spores, is distinctive enough under the light microscope to have value for identification. This means that differential staining techniques, such as the Gram stain or acid-fast stain, and fluorescence microscopy may help to determine the iden-... 

Noise can be also introduced by biochemical heterogeneity of the specimen. This can be a major cause of uncertainty in biological imaging. The high (three-dimensional) spatial resolution of fluorescence microscopy results in low numbers of fluorophores in the detection volume. In a typical biological sample, the number of fluorophores in the detection volume can be as low as 2-3 fluorophores for a confocal microscope equipped with a high NA objective at a fluorescent dye concentration of 100 nM. This introduces another source of noise for imaging applications, chemical or molecular noise, related to the inherent randomness of diffusion and the interaction of molecules. 

SPMs such as AFM, FFM, and SSPM were performed with a SEIKO SPA-300 unit together with an SPI-3700 control station. Details for fluorescence microscopy by SNOAM based on a modified SEIKO SPA-300 unit with an SPI-3700 control station were reported previously [27-33]. Conventional fluorescence microscopy was carried out with a Nikon XF-EFD2 fluorescence microscope [40]. SIMS was performed with a Perkin Elmer PHI model 6600 SIMS system with a Ga liquid metal ion source (beam diameter ca. 80 nm). For mapping of F negative secondary ions, a width of ca. 50 pm was scanned with 256 lines. 

Fluorescence microscopy of phase separated mixed monolayer by a convnetional fluorescence microscope and SNOAM... 

Plant Cells and Tissues Structure-Function Relationships. Methods for the Cytochemical/Histochemical Localization of Plant Cell/Tissue Chemicals. Methods in Light Microscope Radioautography. Some Fluorescence Microscopical Methods for Use with Algal, Fungal, and Plant Cells. Fluorescence Microscopy of Aniline Blue Stained Pistils. A Short Introduction to Immunocytochemistry and a Protocol for Immunovi-sualization of Proteins with Alkaline Phosphatase. The Fixation of Chemical Forms on Nitrocellulose Membranes. Dark-Field Microscopy and Its Application to Pollen Tube Culture. Computer-Assisted Microphotometry. Isolation and Characterization of... 

Microscopic techniques, 70 428 Microscopists, role of, 76 467 Microscopy, 76 464-509, See also Atomic force microscopy (AFM) Electron microscopy Light microscopy Microscopes Scanning electron microscopy (SEM) Transmission electron microscopy (TEM) acronyms related to, 76 506-507 atomic force, 76 499-501 atom probe, 76 503 cathodoluminescence, 76 484 confocal, 76 483-484 electron, 76 487-495 in examining trace evidence, 72 99 field emission, 76 503 field ion, 76 503 fluorescence, 76 483 near-held scanning optical,... 

A new suspension array concept based on sedimentation and microscopic imaging was introduced by Moser et al. [98], Magnetic microbeads settle to the bottom of a microplate well by magnetic forces and form randomly ordered arrays, which are examined by fluorescence microscopy and automated imaging analysis. Each bead carries specific capture molecules and can be identified by a defined luminescent code. 

The research group led by Dr. Djilali at the University of Victoria has developed an ex situ experimental technique using fluorescent microscopy to study the liquid water transport mechanisms inside diffusion layers and on their surfaces [239-243]. The diffusion layer is usually placed between two plates (the top plate may or may not have a channel) the liquid water, which is pumped through a syringe pump, flows from the bottom plate through the DL. Fluorescein dye is added to the water for detection with the microscope. 

Combining fluorescence spectroscopy with fluorescence microscopy, confocal microscopy could be used to elucidate the pathway of siderophore-mediat iron uptake in the fungus Ustilago maydis, and visualize this pathway by providing unique fluorescent microscopic images. Using these techniques, clear images of two independent iron-uptake mechanisms have become visualized as well as their cellular compartment locahzed. 

The time-resolved detection of fluorescence microscopy has been achieved conventionally by these two methods. Therefore, the time resolution of the fluorescence microscope was limited to several tens of picoseconds or nanoseconds (Krishnan et al. 2003 McLoskey et al. 1996). 

The time-resolved techniques that are usually used for FLIM are based on electronic-basis detection methods such as the time-correlated single photon counting or streak camera. Therefore, the time resolution of the FLIM system has been limited by several tens of picoseconds. However, fluorescence microscopy has the potential to provide much more information if we can observe the fluorescence dynamics in a microscopic region with higher time resolution. Given this background, we developed two types of ultrafast time-resolved fluorescence microscopes, i.e., the femtosecond fluorescence up-conversion microscope and the... 

Greenhalgh, C., Cisek, R., Prent, N., Major, A., Aus der Au, J., Squier, J., and Barzda, V. 2005. Time and structural image analysis of microscopic volumes, simultaneously recorded with second harmonic generation, third harmonic generation, and multiphoton excitation fluorescence microscopy. Proc. SPIE 5969 59692F1-F8. 

In this chapter we explore several aspects of interferometric nonlinear microscopy. Our discussion is limited to methods that employ narrowband laser excitation i.e., interferences in the spectral domain are beyond the scope of this chapter. Phase-controlled spectral interferometry has been used extensively in broadband CARS microspectroscopy (Cui et al. 2006 Dudovich et al. 2002 Kee et al. 2006 Lim et al. 2005 Marks and Boppart 2004 Oron et al. 2003 Vacano et al. 2006), in addition to several applications in SHG (Tang et al. 2006) and two-photon excited fluorescence microscopy (Ando et al. 2002 Chuntonov et al. 2008 Dudovich et al. 2001 Tang et al. 2006). Here, we focus on interferences in the temporal and spatial domains for the purpose of generating new contrast mechanisms in the nonlinear imaging microscope. Special emphasis is given to the CARS technique, because it is sensitive to the phase response of the sample caused by the presence of spectroscopic resonances. 

Figure 9.29 Membrane formation by meteoritic amphiphilic compounds (courtesy of David Deamer). A sample of the Murchison meteorite was extracted with the chloroform-methanol-water solvent described by Deamer and Pashley, 1989. Amphiphilic compounds were isolated chromatographically on thin-layer chromatography plates (fraction 1), and a small aliquot ( 1 p,g) was dried on a glass microscope slide. Alkaline carbonate buffer (15 p,l, 10 mM, pH 9.0) was added to the dried sample, followed by a cover slip, and the interaction of the aqueous phase with the sample was followed by phase-contrast and fluorescence microscopy, (a) The sample-buffer interface was 1 min. The aqueous phase penetrated the viscous sample, causing spherical structures to appear at the interface and fall away into the medium, (b) After 30 min, large numbers of vesicular structures are produced as the buffer further penetrates the sample, (c) The vesicular nature of the structures in (b) is clearly demonstrated by fluorescence microscopy. Original magnification in (a) is x 160 in (b) and (c) x 400.

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