Basic confocal laser scanning

Special emphasis is placed on the carbohydrate-mediated cell - target system interaction by describing hints and pitfalls of assays for cytoadhesion, specificity, cytoinvasion, and cytoevasion. In addition, basic considerations are presented to discriminate between active and passive uptake as well as to detect lysosomal accumulation. Finally, the pros and cons of two useful analytical techniques, namely, flow cytometry and confocal laser scanning microscopy, are described in detail. 

The basic optical pathway for the confocal microscope is shown in Fig. 1 (redrawn in part from Carl Zeiss The Confocal Laser Scanning Microscope). This schematic diagram represents the optical pathway and image information flow of a confocal laser scanning microscope. All principle components of a generic CLSM are labeled. 

The APMS used for this separation had an average particle size of 4-10 pm Normal phase HPLC of ferrocene and acetylferrocene performed with non-porous 1-3 pm spheres prepared in basic solution showed only one broad peak with no separation of the target molecules. Similarly, 20 pm spheres prepared in acidic solution showed no resolution of the ferrocenes (Figure 1). This indicates that particle size has some effect on the quality of the HPLC separation, but surface area is the major factor provided that the molecules to be separated can access the interiors of the mesoporous particles, which is dependent upon the pore size. (Experiments performed on APMS using confocal scanning laser microscopy indicated that these particles are porous throughout their interiors). 

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