Inverted microscope

Figure C 1.5.13. Schematic diagram of an experimental set-up for imaging 3D single-molecule orientations. The excitation laser with either s- or p-polarization is reflected from the polymer/water boundary. Molecular fluorescence is imaged through an aberrating thin water layer, collected with an inverted microscope and imaged onto a CCD array. Aberrated and unaberrated emission patterns are observed for z- and xr-orientated molecules, respectively. Reprinted with pennission from Bartko and Dickson [148]. Copyright 1999 American Chemical Society.

For studying settled materials in liquids, or for very large opaque specimens, the inverted microscope may be used. 

With regard to the confinement and enhancement ability of a metallic nano-tip, we have proposed near-field Raman microscopy using a metallic nano-tip [9]. The metallic nano-tip is able to enhance not only the illuminating light but also the Raman scattered light [9, 15, 16]. Figure 2.5 illustrates our nano-Raman microscope that mainly comprises an inverted microscope for illumination and collection of Raman... 

Basic equipment for plant culture media preparation and sterilization, controlled growth chamber, microscope, stereomicroscope and inverted microscope equipped with a photocamera, laminar flow cabinet. 

Material required Stock solution of 25 mM H2DCFDA, carboxy-2, 7 -dichlorofluorescein diacetate (Sigma Co) dissolved in dimethyl sulphoxide (DMSO). Confocal Microscope BIORAD 1024, Laser KrAr with inverted microscope Nikon TMD 300. 

Various optical detection methods have been used to measure pH in vivo. Fluorescence ratio imaging microscopy using an inverted microscope was used to determine intracellular pH in tumor cells [5], NMR spectroscopy was used to continuously monitor temperature-induced pH changes in fish to study the role of intracellular pH in the maintenance of protein function [27], Additionally, NMR spectroscopy was used to map in-vivo extracellular pH in rat brain gliomas [3], Electron spin resonance (ESR), which is operated at a lower resonance, has been adapted for in-vivo pH measurements because it provides a sufficient RF penetration for deep body organs [28], The non-destructive determination of tissue pH using near-infrared diffuse reflectance spectroscopy (NIRS) has been employed for pH measurements in the muscle during... 

As described before, special built-in equipment is not always necessary for dark-field microscopy, and one can readily convert bright-field illumination to dark-field illumination. An inverted microscope equipped with long-distance (low NA) lenses is suitable for setup of a dark-fieldmicroscope. A handmade ring-slit is available when objective lenses such as x4 lens withNA 0.13, xlO lens withNA 0.30, and x20 lens... 

Inverse spinels, 11 60, 61 Inversion temperatures, 25 307 Inverted bell-type pressure element, 20 647 Inverted microscope, 16 471 Inverting filter centrifuge, operation of, 5 545... 

TIRF is easy to set up on a conventional upright or inverted microscope with a laser light source or, in a special configuration, with a conventional arc source. TIRF is completely compatible with standard epi-fluorescence, bright-field, dark-field, or phase contrast illumination so that these methods of illumination can be switched back and forth readily. Some practical optical arrangements for observing TIRF through a microscope are described in Section 7.4. 

Other TIRF configurations for inverted microscopes have been employed. Figure 7.10 shows an alternative system.(42) Instead of a prism fixed with respect to the beam as above, the prism is fixed with respect to the sample. The glass slide substrate propagates the incident beam toward the... 

Figure 7.8. TIRF optical configuration for an inverted microscope detail of the sample chamber region.

TIRF is an experimentally simple technique for selective excitation of fluorophores on or near a surface. It can be set up on a standard upright or inverted microscope, preferably but not necessarily with a laser source, or in a nonmicroscopic custom setup or commercial spectrofluorimeter. In a microscope, the TIRF setup is compatible and rapidly interchangeable with bright-field, dark-field, phase contrast, and epi-illumination and accommodates a wide variety of common microscope objectives without alteration. 

The laser beam can be directed along one of several optical paths available in either the upright or inverted microscope configurations. For fluorescence applications, it is often convenient to make use of the standard epi-illumination... 

Fig. 7. Schematic diagram of forces exerted on a cell when using an inverted microscope with (A) epi-illumination (i.e., laser focused through the objective) or (B) transillumination (i.e., laser focused through the condenser). is the axial force, and Fl is the lateral trapping force. Curved arrows represent the laser beam waist and point in the direction of light propagation.

Image the plate (see Note 16) at intervals starting from t= 0 up to 72 h, using an inverted microscope (see Note 17). 

The laser light travels through the epifluorescence or side port of the microscope. A dichroic mirror reflects the laser light and passes the green fluorescence to either of the detectors. Detectors are positioned on the bottom port of the inverted microscope or the top port of the upright microscope. The choice of detector is discussed in more detail below. Broadband and band-pass filters placed in the detection path prevent residual IR from reaching either of the detectors. 

Figure 10.10 shows the experimental system of TE-CARS microscopy (Ichimura et al. 2004a). As similar to the TERS system (Hayazawa et al. 2000), the system mainly consists of an excitation laser, an inverted microscope, an AFM using a silver-coated probe, and a monochromator. Two mode-locked Ti sapphire lasers (pulse duration 5 picoseconds [ps] spectral band width 4 cm- repetition rate 80 MHz) are used for the excitation of CARS. The (o and (O2 beams are collinearly combined in time and space, and introduced into the microscope with an oil-immersion objective lens (NA = 1.4) focused onto the sample surface. As the z-polarized component of the... 

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