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Download Confocal Raman Microscopy by Thomas Dieing, Olaf Hollricher, Jan Toporski PDF

By Thomas Dieing, Olaf Hollricher, Jan Toporski

Confocal Raman Microscopy is a comparatively new process that permits chemical imaging with no particular pattern education. by way of integrating a delicate Raman spectrometer inside a state of the art microscope, Raman microscopy with a spatial answer all the way down to 200nm laterally and 500nm vertically could be accomplished utilizing noticeable mild excitation. fresh advancements in detector and laptop expertise in addition to optimized tool layout have lowered integration occasions of Raman spectra via orders of importance, in order that whole photographs inclusive of tens of hundreds of thousands of Raman spectra should be received in seconds or mins instead of hours, which was ordinary only one decade in the past. the aim of this publication is to supply the reader a entire review of the quickly constructing box of Confocal Raman Microscopy and its applications.

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Struct. J. Thomas, Annu. Rev. Biophys. Biomol. Struct. 28, 1 (1999) R. Withnall, Protein-Ligand Interactions, Structure and Spectroscopy: A Practical Approach, chap. Protein-Ligand Interactions Studied by Raman and Resonance Raman Spectroscopy, (Oxford University Press, Oxford, New York, 2001). K. Gaus, P. Rösch, R. D. Peschke, O. Ronneberger, H. Burkhardt, K. Baumann, J. M. R. Goodacre, FEMS Microbiol. Lett. C. Lopez-Diez, R. Goodacre, Anal. Chem. Q. A. J. Thomas, Biochemistry 36, 7810 (1997) M.

4. In case resonance Raman spectroscopy occurs via the same electronic excited states where the fluorescence emission originates (Fig. 4a), the Raman signals are often masked by the broad and featureless fluorescence spectra. , the energy difference between the absorption maximum and the fluorescence maximum, is only small (Fig. 4c). In UV resonance Raman spectroscopy, energy-rich UV light is used, so that higher electronic excited states are incorporated. Raman scattering occurs within the first few femtoseconds (10−15 s) directly from the high electronic state.

If G is the object size and M the magnification of the objective, then the lateral image size is given by B = G · M. But what about the axial magnification? 17) Without the diffraction limit, an object of an axial extension of 200 nm would have a length of 2 mm in the image space at a magnification of M = 100. 17), the depth of focus is very high and the axial position of the pinhole is not critical. The choice of the pinhole size is very important in Raman experiments. On one hand the signal should be as high as possible, while on the other hand the image should be confocal.

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