Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

作者：Cseresnyes Zoltan; Oehme Laura; Andresen Volker; Sporbert Anje; Hauser Anja E^*; Niesner Raluca

来源：Jove-Journal of Visualized Experiments, 2014, (86): e51135.

DOI：10.3791/51135

• Immunology
• Issue 86
• two-photon laser scanning microscopy
• deep-tissue intravital imaging
• germinal center
• lymph node
• high-resolution
• enhanced contrast

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 mu m 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.

• 出版日期2014-4