%0 Journal Article %1 Sijtsema1998 %A Sijtsema, Nanna M. %A Wouters, Siddi D. %A Grauw, Cees J. De %A Otto, Cees %A Greve, Jan %D 1998 %J Appl. Spectrosc. %K hardware microscopy raman %N 3 %P 348-355 %R 10.1366/0003702981943806 %T Confocal Direct Imaging Raman Microscope: Design and Applications in Biology %V 52 %X A confocal direct imaging Raman microscope (CDIRM) based on two synchronized scanning mirrors, a monochromator, and two charge-coupled device (CCD) cameras has been developed. With this system it is possible to make both Raman spectra of a small measurement volume and images of a larger sample area in one speci® c Raman band. The spatial resolution of the system was determined for two limiting situations: a small sphere and a thin layer. The image of a 0.282 m m sphere appeared to have a full width at half-maximum (FWHM) of 1.2 m m in the axial and 0.37 m m in the lateral direction, whereas the image of a 275 nm layer showed an FWHM of 1.4 m m in the axial direction . Confocal Raman images were made of the DNA and protein distribution in polytene chromosomes with a relatively weak Raman signal 0.1 photons/(second pixel). Further, a three-dimensional Raman image of the drug distribution in a phthalocyanine-incubated fixed cell is presented. These examples show that the CDIRM can be used to image samples with a weak Raman signal and that three-dimensional images of the distribution of specific molecules in a sample can be made.

@article{Sijtsema1998, abstract = {A confocal direct imaging Raman microscope (CDIRM) based on two synchronized scanning mirrors, a monochromator, and two charge-coupled device (CCD) cameras has been developed. With this system it is possible to make both Raman spectra of a small measurement volume and images of a larger sample area in one speci® c Raman band. The spatial resolution of the system was determined for two limiting situations: a small sphere and a thin layer. The image of a 0.282 m m sphere appeared to have a full width at half-maximum (FWHM) of 1.2 m m in the axial and 0.37 m m in the lateral direction, whereas the image of a 275 nm layer showed an FWHM of 1.4 m m in the axial direction . Confocal Raman images were made of the DNA and protein distribution in polytene chromosomes with a relatively weak Raman signal [0.1 photons/(second pixel)]. Further, a three-dimensional Raman image of the drug distribution in a phthalocyanine-incubated fixed cell is presented. These examples show that the CDIRM can be used to image samples with a weak Raman signal and that three-dimensional images of the distribution of specific molecules in a sample can be made.}, added-at = {2010-03-09T22:47:53.000+0100}, author = {Sijtsema, Nanna M. and Wouters, Siddi D. and Grauw, Cees J. De and Otto, Cees and Greve, Jan}, biburl = {https://www.bibsonomy.org/bibtex/2290bff197c288bfc8618e88b01341f14/afcallender}, doi = {10.1366/0003702981943806}, file = {Sijtsema1998.pdf:indexed\\Sijtsema1998.pdf:PDF}, interhash = {74bda91fc1527a8023e200bce0330ba8}, intrahash = {290bff197c288bfc8618e88b01341f14}, journal = {Appl. Spectrosc.}, keywords = {hardware microscopy raman}, number = 3, pages = {348-355}, timestamp = {2010-03-09T22:59:01.000+0100}, title = {Confocal Direct Imaging Raman Microscope: Design and Applications in Biology}, volume = 52, year = 1998 }