Epi fluorescence microscope

Epi-fluorescence microscope equipped with high pressure mercury light source using a 580 nm dichromic mirror, 510-560 nm excitation filter and 590 nm barrier filter for propidium iodide staining (e.g., Nikon inverted microscope Diaphot model TMD). 

A. Single-Molecule Imaging with an Epi-Fluorescence Microscope... 

Here these techniques will be discussed from a general point of view and instructive examples will be presented. For didactic reasons, single-molecule imaging with a commercial epi-fluorescence microscope will be discussed... 

Specimens for microscopy were obtained from the bonded specimens and from the samples after the D 905 tests. For transverse sections, samples were water-soaked for 2-24 h prior to microtoming. The sections were analyzed using a scanning electron microscope (JEOL 840 after gold plating the samples) or a Leitz Orthoplan epi-fluorescence microscope with a 150-W mercury lamp light source, an A2 UV filter cube, and a Nikon DS-5M digital camera, or a Carl Zeiss Axioskop epi-fluorescence microscope with a 100-W mercury lamp, a UV filter set, and CCD camera. Fluorescence microscopy was used to examine the failure surface. 

The objective-type TIRFM is based on the fact that a laser beam off-axis at the BFP of the objective leaves the objective lens with an angle. When the incident beam is focused at the center of the objective on-axis, the microscope can be used as a standard EPI-fluorescence microscope. When the path is shifted from the center to the edge off-axis (i.e., between... 

Visualization of flow illustrates the pattern of flow in a designed microchaimel geometry which helps in understanding device operation and predicting the particle (and ultimately cell) motion. To visualize flow, fluorescent tracer particles of diameter < 1 pm are used. Tracer particles with such a small diameter have very limited inertial effect on them thus, these particles are evenly dispersed in the flow forming particle cloud. By using an inverted epi-fluorescence microscope, the flow pattern can be observed. Using the vortex-based crmtinuous cell extraction device as example [9], the process... 

Fig. 4 Schematic of a typical experimental setup used to carry out p-PIV measurements. A pulsed Nd YAG laser or a continuous Hg arc lamp is used to excite fluorescently tagged polystyrene particles on an inverted epi-fluorescent microscope. A CCD camera is used to capture the particle images at regular well-defined time intervals. A filter cube is selected based on the fluorophore used (i.e., FITC for fluorescein or TRITC for rhodamine)...

Disadvantages An expensive epi-fluorescent microscope is required to visualize the in situ hybridization signal and the signal is unstable,... 

With an epi-fluorescent microscope, a dry objective lens (40x, NA = 0.75) and fluorescent particles with a diameter dp = 500 nm, a single-camera p-PTV technique using the described deconvolution microscopy has been developed by Park and Kihm [31]. In this case, only the outermost diffraction pattern ring size variations of defocused particle images are compared with a computed PSF. With this technique, it is possible to map simultaneously the three-component velocity vectors for 3D microscale flow fields. The measured 3D vector fields for the creeping flow over a 95 pm diameter sphere inside a nominal 100 pm square channel is shown in Figure 4.9. 

Adachi, K, Kinosita Jr, K, and Ando, T, Single-fluorophore imaging with an unmodified epi-fluorescence microscope and conventional video camera. Journal of Microscopy 195 (Pt 2) (1999) 125-132. 

A fluorescence microscope setup that uses such optical filter set, as depicted in Fig. 2, is known as an Epi-Fluorescence Microscope [1]. The word Epi-Fluorescence means that both the excitation light path and the emission light path use the same lens near the fluorochrome molecules. When excitation light illuminates the fluorochrome molecules, the lens serves as a condenser lens, which focuses the light into the object plane. Mean-... 

Figure 10 illustrates schematic drawing of objective-type TIRFM. A high numerical aperture objective lens is mounted on an inverted microscope. A laser beam is passed through a neutral density filter (ND) and a beam expander (BE) to adjust its power and diameter. When the laser polarized linearly is used, the polarization of the laser beam is converted from linear to circular by a quarter-wave plate (A/4). The laser beam is focused by a lens (L) on the back focal plane of the objective, so that specimens are illuminated uniformly with Koehler illumination. By shifting the position of the mirror (M) located between the lens (L) and dichroic mirror (DM), the path of the incident laser light is shifted from the center to the edge of the objective. At the center position, the microscope can be used as a standard epi-fluorescence microscope (Fig. 10b). 

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