Multicolor microscopy

The recent work at the Polymer Branch at US Air Force Wright Laboratory and that at Photonics Research Laboratory at SUNY, Buffalo have produced organic structures that exhibit strong two- and three-photon absorption and exhibit strong up-converted fluorescence. Furthermore, we have shown that with the same pump source for multiphoton excitation, one can excite a number of different fluorophores to generate multicolors to conduct multiphoton multicolor microscopy for the study of microstructures of multilayer coatings, their interfaces and bulk as a function of depth. 

Shino A, Lujioka K, Manabe N, Yamaya S, Goto Y, Yasuhara M, Yamamoto K (2005). Simultaneous multicolor detection system of the single-molecular microbial antigen with total internal reflection fluorescence microscopy. Microbiol. Immunol. 49 461 —170. 

Wu, Y., Campos, S. K., Lopez, G. P., Ozbun, M. A., Sklar, L. A., and Buranda, T. (2007b). The devolpment of quantum dot calibration beads and quantitative multicolor bioassays in flow cytometry and microscopy. Anal. Biochem. 364, 180-192. 

L. Schermelleh, P.M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M.C. Cardoso, D.A. Agard, M.G.L. Gustafsson, et ah. Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy. Science 320, 1332-1336 (2008)... 

For living-cell microscopy, multilabeling acquisition is constrained by the fact that objects are likely to move during the collection of stacks. Multicolor images must therefore be acquired as rapidly as possible, and the sequential stack-color mode described above is inappropriate. There is a single solution to this problem using a multiband pass dichroic mirror and dissociating... 

The spatial variations of the InGaN thickness and composition may lead to multicolor emissions. Figure 14.6 shows microscopic photoluminescence (PL) spectra acquired at room temperature (RT) using a confocal microscopy system. The spatial resolution of the system is less than 500 run. Hence, each... 

Transfection techniques in vitro and ex vivo (organotypic cultures) offer an array of possibilities to investigate the consequence) s) of the introduction of foreign nucleic acids (DNA but also RNA) and or other biologically active molecules into neurons, and to combine observations with immunocytochemistry. In particular, a wide number of fluorescent reporter proteins (FRPs) can be employed for multicolor fluorescence imaging. Here we present a series of protocols for in vitro and ex vivo transfection of DNA or RNA sequences into cerebellar neuron cultures and organotypic slices based on the use of plasmid vectors and multicolor laser scanning confocal microscopy. These protocols allow analysis of live transfected cells, and, after fixation, correlative neurochemical studies. 

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