title: Confocal laser scanning microscopy as an analytical tool in chromatographic research.
abstract: In recent years, confocal laser scanning microscopy has been developed into a non-invasive tool to probe intra-particle profiles of protein in chromatographic adsorbents. A necessary prerequisite when using this technique lies in the labeling of proteins with fluorescent probes. The quality of the obtained results is thus strongly dependent on the probes used, its sensitivity on experimental parameters and the change of protein characteristics upon binding. In this review, the fundamental issues when using fluorescent probes are described, before giving a critical evaluation on published literature in the field of confocal laser scanning microscopy for the analysis of chromatographic principles.
authors.affiliation: Institut für Biotechnologie 2, Forschungszentrum Jülich, 52425 Jülich, Germany. j.hubbuch@fz-juelich.de
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hubbuch
authors.forename: Maria Regina
authors.initials: MR
authors.lastname: Kula
mesh_terms: D000327:Adsorption; D000465:Algorithms; D001709:Biotechnology / Q000379:methods; D002845:Chromatography / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry; D018613:Microscopy, Confocal / Q000379:methods*; D008968:Molecular Conformation; D038761:Photobleaching; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D011506:Proteins