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Highly resolved intravital striped-illumination microscopy of germinal centers

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Item Type:Article
Title:Highly resolved intravital striped-illumination microscopy of germinal centers
Creators Name:Cseresnyes, Z. and Oehme, L. and Andresen, V. and Sporbert, A. and Hauser, A.E. and Niesner, R.
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.
Keywords:Two-Photon Laser Scanning Microscopy, Deep-Tissue Intravital Imaging, Germinal Center, Lymph Node, High-Resolution, Enhanced Contrast, Animals, Mice
Source:Journal of Visualized Experiments
ISSN:1940-087X
Publisher:JoVE (U.S.A.)
Number:86
Page Range:e51135
Date:9 April 2014
Official Publication:https://doi.org/10.3791/51135
PubMed:View item in PubMed

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