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| Item Type: | Article |
|---|---|
| Title: | Highly resolved intravital striped-illumination microscopy of germinal centers |
| Creators: |
Cseresnyes, Z.  ORCID: https://orcid.org/0000-0002-6574-2588, Oehme, L., Andresen, V., Sporbert, A. ORCID: https://orcid.org/0000-0002-3738-1295, Hauser, A.E. and Niesner, R. ORCID: https://orcid.org/0000-0001-8969-5432
|
| 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 |
| 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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