Imaging of Immune Cells : In vitro veritas?

✍ Scribed by Matthias Gunzer; Bastian Dornbach

• Publisher: Wiley (John Wiley & Sons)
• Year: 2006
• Weight: 630 KB
• Volume: 8
• Category: Article
• ISSN: 1439-4243
• DOI: 10.1002/imic.200790090

✦ Synopsis

In vitro veritas?

High resolution microscopic imaging is an important technology to study the physical interactions of immune cells. The immune system comprises a complex network of different cell types with the task to defend and protect the host organism from infection or cancer. We observe those interactions to receive a better understanding of the complex molecular interrelations which lead to a functional immune response (e.g. activation of cells, antibody production, killing events). Therefore, we established three methods to image and analyse interaction events: first, immunofluorescence stainings of fixed samples, second, in vitro live cell imaging in a collagen matrix and finally, in vivo live cell imaging in a mouse lymph node. Thus, three distinct technological approaches lead to a mosaic-like composed picture of how immune cells behave.

Methods

We use the transgenic DO11.10 mouse line, whose T cell receptor (TCR) recognises a peptide of chicken ovalbumin presented by BALB/c APCs. Cells are obtained from spleens of mice using a magnetic cell sorting system (Miltenyi).

Fixed cell samples are received by adhering interacting cells on Poly-L-Lysine