TY  - JOUR
TI  - Imaging Thick Lymph Node Tissue Sections
AU  - Hor, Jyh Liang
AU  - Mueller, Scott N.
VL  - 6
IS  - 18
PY  - 2016
DA  - 2016/09/20
SP  - e1938
C1  - Bio-protocol 2016;6:e1938
DO  - 10.21769/BioProtoc.1938
UR  - https://doi.org/10.21769/BioProtoc.1938
AB  - Our protocol describes a simple procedure for imaging thick lymph node sections by 2-photon microscopy. Lymph nodes are sectioned using a vibratome (vibrating microtome) to produce slices of tissue that can then be stained with fluorescently labeled antibodies. The thick tissue sections (150-200 μm depth) allow for the detection of cell clustering that is typically under-represented in thin sections (10-20 μm) used for conventional confocal microscopy. Application of 2-photon microscopy facilitates imaging through the thick volume of the vibratome sections. In combination with automated image processing software, a thick lymph node cross-section image also facilitates quantitation of cellular events within a relatively large area of the tissue, thus providing a clearer picture on the spatial distribution of cellular events of interest (e.g., T cell clustering). This method can also readily be applied to other tissues, such as the spleen or skin.
KW  - 2-photon microscopy
KW  - Lymph nodes
KW  - T lymphocytes
KW  - Vibratome
KW  - Immune response
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False