title: Highly resolved intravital striped-illumination microscopy of germinal centers.
abstract: Monitoring cellular communication by intravital deep-tissue multi-photon microscopy is the key for understanding the fate of immune cells within thick tissue samples and organs in health and disease. By controlling the scanning pattern in multi-photon microscopy and applying appropriate numerical algorithms, we developed a striped-illumination approach, which enabled us to achieve 3-fold better axial resolution and improved signal-to-noise ratio, i.e. contrast, in more than 100 µm tissue depth within highly scattering tissue of lymphoid organs as compared to standard multi-photon microscopy. The acquisition speed as well as photobleaching and photodamage effects were similar to standard photo-multiplier-based technique, whereas the imaging depth was slightly lower due to the use of field detectors. By using the striped-illumination approach, we are able to observe the dynamics of immune complex deposits on secondary follicular dendritic cells - on the level of a few protein molecules in germinal centers.
authors.affiliation: Biophysical Analytics, German Rheumatism Research Center, Leibniz Institute; Microscopy Core Facility, Max-Delbrück Center for Molecular Medicine.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Cseresnyes
authors.affiliation: Immunodynamics, German Rheumatism Research Center, Leibniz Institute.
authors.forename: Laura
authors.initials: L
authors.lastname: Oehme
authors.affiliation: LaVision Biotec GmbH.
authors.forename: Volker
authors.initials: V
authors.lastname: Andresen
authors.affiliation: Microscopy Core Facility, Max-Delbrück Center for Molecular Medicine.
authors.forename: Anje
authors.initials: A
authors.lastname: Sporbert
authors.affiliation: Immunodynamics, German Rheumatism Research Center, Leibniz Institute; Immunodynamics and Intravital Imaging, Charité - University of Medicine; hauser@drfz.de.
authors.forename: Anja E
authors.initials: AE
authors.lastname: Hauser
authors.affiliation: Biophysical Analytics, German Rheumatism Research Center, Leibniz Institute; niesner@drfz.de.
authors.forename: Raluca
authors.initials: R
authors.lastname: Niesner
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D018858:Germinal Center / Q000166:cytology* / Q000276:immunology; D049452:Green Fluorescent Proteins / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*
chemical_list: D000936:Antigen-Antibody Complex; D049452:Green Fluorescent Proteins
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