Confocal fluorescence microscope

In this chapter, it was shown that filterFRET is an easy, intuitive and quantitative alternative to record sensitized emission and FRET efficiency. The major advantages of filterFRET over donor-based FRET detection methods (FLIM) are that it can be carried out with standard wide-held or confocal fluorescence microscopes that are available in most laboratories, and that it yields additional data on the acceptor population. FilterFRET is also fast, requiring just two confocal scans (if need be on a line-by-line basis) which minimizes the risk of artifacts due to, for example, organelle movement in living cells, and acquisition can be optimized for each channel independently. However, quantitative... 

Fig. 12 CNTs act as a vector for drug delivery into living cells. After incubation of HeLa cells with AlexaFluor594-labeled SWNTs for 12 h at 37 °C, living cells were observed under confocal fluorescence microscope for a CNT uptake study, (a) Images show dual confocal detection of AlexaFluor594-SWNT (red) internalized into cells with the membrane stained by AlexaFluor488 (green), (b) Series of images of different z-focal scanning planes down through cells. (Adapted from [61])...

BT 20 cells incubated in serum free medium for 10 hours with the vector/ DNA complexes (DQAplexes, C-DQAplexes). For control, cells were exposed to naked DNA and empty vesicles. The cells were then stained with Mitotracker Red CMXRos (Molecular Probes) for five minutes to enable the visualization of mitochondria followed by confocal fluorescence microscopic analysis on a Zeiss Meta 510 Laser Scanning Microscope. 

Figure 11.21 Confocal fluorescence microscope image showing the cellular localisation behaviour of the europium complex 11.33 in living cells, revealing the staining of the nucleolus inside the cell nucleus (reproduced by permission of The Royal Society of Chemistry). 

P 66] Equal streams of 1 mM solutions of fluorescein-labeled polymer (polyethylen-imine, molecular weight 500 000) in water-glycerol mixtures (0 and 80% glycerol) and a clear solution were injected into the channel [44], The flow was achieved by compressed air at constant pressure. Imaging was achieved by applying a confocal fluorescence microscope. 

Dai Y, Whittal R, Li L (1996) Confocal fluorescence microscopic imaging for investigating the analyte distribution in MALDI matrices. Anal Chem 68 2494-2500... 

For analysis of the uptake of fluorescent-labeled molecules flow cytometer or ELISA fluorescent plate reader, confocal fluorescent microscope are used. 

Fig. 2.10. Electron microscope images of sihca particles (a) 300 nm CP-grafted particle (TEM) (b) 5 pm CP-grafted particle (SEM) (c) Confocal fluorescence microscope image of 5 pm CP-grafted particies. Reproduced from ref. [55] with permission. Copyright 2007, American Chemicai Society...

Wash each coverslip twice in 0.1% saponin-PBS, once in PBS, and finally once in water to remove salts. Drain all liquid onto a Kimwipe and mount coverslips on microscopy slides by inverting them (cells facing down) on 10-pL drops of Mowiol mounting medium. Allow at least 2 h of polymerization before viewing samples on a confocal fluorescence microscope equipped with 488-, 568-, and 643-nm laser lines. 

Imaging by confocal fluorescence microscope and correcting for photobleach-ing and the smoothing are essentially done as described for the model system (Section II,B). Usually the imaging of a particular nucleus is restricted to its upper sperical shell. From 11 to 16 xy images, separated in the z direction by 0.2 jum, are acquired, thus covering a total volume of 20 /.im x 20 ixm X 2-3 (xm. 

Fig. 2.18 A Confocal fluorescence microscope images of the QDs on (Aa) a flat gold film, (Ab) the conical Au array, and (Ac) the dimpled Au array. (Ad) 3D visualization of the fluorescence intensity in (Ac) [60]. The QD has an emission wavelength of 597 nm. The excitation laser source was a 543 nm HeNe laser. (B) Field snapshot image from FDTD simulation results of Au coated dimpled structure. (C) Plots for the near-zone field intensity versus time in the dimpled Au structure [60]. Reproduced with permission [60]. Copyright 2013, Royal Society of Chemistry...

S.W. HeU, E. Stelzer, Eundamental improvement of resolution with a 4 Iri-confocal fluorescence microscope using two-photon excitation. Opt. Commun. 93, 277 (1992) ... 

Fluorescence Measurements, Fig. 3 The schematic concept of a confocal fluorescence microscope configuration. LS light source, OD optical detector, ExF excitation filter, EmF emission filter, DM dichroic mirror, L lens, EM position of fluorochrome molecules, LSP light-source pinhole, DP detector pinhole. Note that the emission light coming Irom out-of-focus fluorochrome molecules is blocked away by the detector pinhole... 

Figure 11.7 Confocal fluorescence microscope image of gold nanoparticles dispersed...

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