Scanning confocal fluorescence microscopy

The availability of laser scanning confocal fluorescence microscopy,68 first commercially offered in the 1980s, enabled a major advance in chemical microscopy imaging convenient summaries are available in the Handbook of Biological Confocal Microscopy.69 Confocal fluorescence microscopy works by focusing continuous-wave laser illumination to a diffraction-limited spot in a focal... 

LSCFM laser scanning confocal fluorescence microscopy... 

In OM, the spatial resolution both lateral (parallel to the focal plane) and axial (perpendicular to the focal plane) is always a critical issue. As to the lateral resolution, laser scanning confocal fluorescence microscopy (LSCFM) has a high resolution of 0.42A/NA, where NA is the numerical aperture hence, the use of a large NA objective with blue laser hght allows one to achieve a high resolution of less than 200 nm [3]. Polymer films containing coumarine-labeled latex particles with diameters of 500,220, or 100 nm were examined to estimate the resolution of LSCFM [4]. The measurement of a single latex particle proved that the accuracy of the setup was in the order of 200 nm laterally and 400 nm axially. 

Fluorescence microscopy has also been employed largely for 2D surface imaging. Scanning confocal fluorescence microscopy has been applied to the investigation of subsurface morphology of foams. The general knowledge on the applications of fluorescence microscopy for polymers is rather limited. 

More recently Ghiggino and co-workers(32) have applied laser scanning confocal fluorescence lifetime microscopy to the study of polyvinyl alcohol films containing rhodamine B (650 nm emission) and cresyl violet (632 nm emission). Synchronously pumped dye laser excitation and APD detection were used with optical fiber coupling. A schematic diagram of their apparatus is shown in Figure 12.5. 

Figure 1.46 Two scanning methods in the confocal microscope (a) specimen scanning and (b) laser scanning. (Reproduced with permission from M. Muller, Introduction to Confocal Fluorescence Microscopy, 2nd ed., SPIE Press, Bellingham, Washington. 2006 Michiel Miiller)...

The last two decades have seen tremendous progress in optical microscopy, especially, confocal fluorescence microscopy. Confocal laser-scanning fluorescence... 

The cellular uptake of these 3LNPs was observed using confocal scanning laser fluorescence microscopy. SKOV-3 ovarian cancer cells were cultured with... 

Fig. 10.11 Adsorption of 3LNPs with PKH26 on SKOV-3 cell membrane at pH 7.4 (a, b) and 6.0 (c, d) at 4°C observed with confocal scanning laser fluorescence microscopy. Differential interference contrast (a, c) and red fluorescence channel (b, d)...

Figure 11.9 Schematic illustration of the principles of confocal fluorescence microscopy singlephoton exdted fluorescence occurs along the entire beam path, however the insertion of a pinhole in the focal plane of the objective ensures that fluorescence from outside the focal point of the laser (dashed line) is prevented from reaching the detector. The laser is scanned in theX-K plane to build up an image of the sample for a given depth. Translation of the sample in the Z-direction allows images (optical sections) to be recorded as a function of depth...

Confocal fluorescent microscopy Confocal laser scanning microscopy Layer-by-Layer (LbL) technique Microcapsules... 

Fig. 5.3. Confocal fluorescence microscopy images of 6-CF precipitates (left) forming in the capsules loaded with PSS. Arrows show broken capsules without PSS. Scanning electron microscopy image (right) of CR precipitates formed [56].

Glass slides Biotin Xmax = 560-700 nm Scanning femtosecond pulsed laser, 0-19.75 mW 100 m s", RT, A(W-dimethyl formamide yes Confocal fluorescence microscopy [RIC 13]... 

Optical imaging with collinear optical coherence and confocal fluorescence microscopy (OCM-CFM). This noninvasive imaging technique combines the use of reflection confocal microscopy in tandem with optical coherence microscopy to characterize scaffolds and tissue constructs. It can probe the interior of scaffolds with the same resolution as conventional laser scanning confocal microscopy but with greater sensitivity (P). 

Coley HM, Amos WB, Twentyman PR, Workman P. Examination by laser scanning confocal fluorescence imaging microscopy of the subcel-lular localisation of anthracyclines in parent and multidrug resistant cell lines. Br J Cancer 1993 67(6) 1316-23. 

STED microscopy is a scanning confocal fluorescence imaging technique. As noted above, confocal microscopy illuminates the sample with a focused light, but the size of the focal spot is limited to approximately 200 nm by the diffraction limit. In STED microscopy, a donut-shaped STED spot is overlapped on the excitation spot, as shown in Figure 19.9. The STED beam has a wavelength oflsTED that... 

If both the illumination and detection systems are simultaneously focused onto a spot which is scanned over the specimen, this is called confocal fluorescence microscopy, and has several advantages. Most... 

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