Axial resolution of confocal Raman microscopes: Gaussian beam theory and practice

A straightforward and transparent model, based on Gaussian beam optics, for the axial r0 resolution of a confocal microscope is presented. A confocal Raman microscope was used to determine the axial confocality in practice. The axial response of a thin planar object was measured for three different objectives, two pinhole sizes and a slit. The results show that, in the case of a confocal configuration, the response calculated with the model provides a good prediction of the axial resolution of the confocal microscope

Application of Raman Microspectroscopic and Raman imaging techniques for cell biological studies

Raman spectroscopy is being used to study biological molecules for some three decades now. Thanks to continuing advances in instrumentation more and more applications have become feasible in which molecules are studied in situ, and this has enabled Raman spectroscopy to enter the realms of biomedicine and cell biology [1-5].\ud Here we will describe some of the recent work carried out in our laboratory, concerning studies of human white blood cells and further instrumentational developments

Raman microspectroscopic and Raman imaging:New techniques for cell biological and biomedical research

Door haar grote molecuulspecifiteit kan Ramanspectroscopie gebruikt worden voor het herkennen/aantonen van moleculen in een preparaat. Het is ook mogelijk om de secundaire structuur van biologische macromoleculen zoals DNA en eiwitten te bestuderen en informatie te verkrijgen over interakties tussen moleculen. Dit alles zonder het gebruik van labels of probes. Enkele voorbeelden daarvan worden besproken

Line-scan Raman microspectrometry for biological applications

A high-resolution confocal line-scan Raman microscope was developed for the study of biological samples such as cells and chromosomes. With the illumination of a line on the sample, all the spectral information of the line is recorded. The line-scan microscope was attained by the introduction of one scanning mirror, a slit, and two cylindrical lenses in a standard confocal Raman microscope. The resolution, for an ideal sample, is 0.5 μm in the lateral direction and 3.5 μm in the axial direction. Line-scan experiments were performed to demonstrate the applicability of the instrument for biological samples. Line-scan images of calcium phosphate bone implant coatings and of the banding pattern of polytene chromosomes are presented