The design of a confocal microscope is very similar to a conventional epi-fluorescence microscope except with additional inclusion of confocal optics. The confocal optics consists of a confocal aperture (adjustable pinholes) which is such placed that it is conjugates the image plane (i.e., focal plane) of the objective lens. The diameter of confocal aperture is infinitesimally small, thus PMT detector can only detect light passing through the pinhole. This implies that, light originating in the focal plane of objective lens can only reach to detector. The very small confocal aperture also allows point detection of the sample. As a result, laser are used as excitation source in confocal and image is captured by point-by-point detection across the whole sample (i.e., raster imaging). Due to exclusion of light from non-focal planes and point detection, highly contrast images are generated in confocal microscopes.
Another important aspect of CLSM system is optical sectioning. Due to variable pinhole and laser excitation it is possible to exclusively image a thin optical slice of a thick sample (typically 100 µm). Under optimum conditions, optical slices of as thin as 500 nm are possible to achieve.
Application of Confocal microscope are immense particularly in the field of biological sciences and biotechnology. Confocal microscopy is used in imaging biological cells, organelles and even study of protein/biomolecule distribution within the cell. It is also used for re-construction of 3D cellular models as well as polymeric networks.
The sample should be transparent and fluorescent in nature. Careful sample preparation step is needed before analysis.
Confocal Imaging - Rs. 1500 Per Sample