Confocal phase-contrast microscope

As described in Section 16.4.5, the detection of refractive-index change between two isomers in a photochromic material is the most promising technique for nondestructive readout.It is necessary to develop a readout system that is sensitive to refractive-index distribution. For this purpose, several methods, such as phase-contrast microscope, differential phase-contrast microscope, and reflection confocal microscope configurations have been proposed. 

21 (a) Readout data by confocai phase-contrast microscope wid (b) its cross section, 

22 Mechanism of the generation of anisotropy by photoisomerizadon of urethane-urea copolymer. 

23 Writing and reading with various polarization directions (a) Readout results of data that were recorded with the polarization angles from 0° to i80° with a pitch of 15°. The data were read out with four polarization states, (b) Polar plot of readout intensity of data with the (unction of the polarization direction of the readout beam. Data were recorded with the horizontally polarized light in this figure, 

24 Readout results of the pt arizadon-multipiexed recorded data at the respective recording polarization angles 0°. 60°, and 120°. 

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A phase contrast wide field transmission microscope combining the advantages of interferomic and confocal techniques has been developed [155]. Confocal operation is achieved by superimposing speckle illumination of a reference beam in a Mach-Zehnder interferometer with a matched speckle pattern of the object beam. The technique was applied to both dry powders and suspensions and gave good agreement with modelled results. Data acquisition time is less than a millisecond. 

The confocal laser-scanning microscopy (Laser-scanning microscope 410 Zeiss) is performed on fresh frozen slices, the excitation wavelength is 488/514 nm of the internal argon laser, the fluorescence detection is performed at >590 nm within the red channel and phase contrast within the green channel (Figure 5a, 5b). 

The vesicles were observed on an inverted microscope in phase-contrast and confocal fluorescence modes, as follows. The white cloud of lipid was gently dispersed in the tube and introduced into an observation chamber. The chamber was filled with the same solution as the internal solution except that 0.1 M sucrose was replaced with 0.1 M glucose (external solution). Because of the difference in the reffactivity of the internal and external solutions, the contrast of the vesicle s images was enhanced in the phase-contrast mode. For the confocal fluorescence microscopy, a confocal scanner unit (CSU 10, Yokogawa, Japan) was used, and the lipophilic fluorescent dye Nile Red (Molecular Probes, Inc., OR, USA) was added to the starting lipid solution at 0.3 wt% of the lipid. 

Microscopy. Particle size, shape and structure of emulsion droplets can be visualized by various microscope techniques, such as phase contrast light microscopy, confocal scanning light microscopy (CSLM), scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray microtomography (XRT), atomic force microscopy (AFM) and imaging techniques. 

Observe the oocytes on the inverted microscope linked to the confocal using either DIG or phase-contrast optics. 

Fig. 13.11 Top Schematic diagram of pH-induced drug release from pH-cleavable diug-loaded supramolecular capsule. CMD-g-p-CD (caiboxymethyl dextran-p-cyclodextrin), PAD-g-AD (polyaldehyde dextran-graft-adamantane), AD-Dox (adamantine-doxonibicin conjugate), Dex-g-FITC (fluorescein isothiocyanate-labeled dextran). Bottom Confocal microscope image of HeLa cells treated with AD-Dox loaded capsules at (a-c) pH 5.5 (al) enlarged image a Dox fluorescence in cells b phase contrast image c overlay fluorescence and phase eontrast image. Scale bar 50 pm. Reproduced from [88] with permission from Ameriean Chemieal Society...

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