In situ imaging of the mouse thymus using 2-photon microscopy.
J Vis Exp
; (11)2008 Jan 25.
Article
em En
| MEDLINE
| ID: mdl-19066556
ABSTRACT
Two-photon microscopy (TPM) enables us to image deep into the thymus and document the events that are important for thymocyte development. To follow the migration of individuals in a crowd of thymocytes , we generate neonatal chimeras where less than one percent of the thymocytes are derived from a donor that is transgenic for a ubiquitously express fluorescent protein. To generate these partial hematopoetic chimeras, neonatal recipients are injected with bone marrow between 3-7 days of age. After 4-6 weeks, the mouse is sacrificed and the thymus is carefully dissected and bissected preserving the architecture of the tissue that will be imaged. The thymus is glued onto a coverslip in preparation for ex vivo imaging by TPM. During imaging the thymus is kept in DMEM without phenol red that is perfused with 95% oxygen and 5% carbon dioxide and warmed to 37 degrees C. Using this approach, we can study the events required for the generation of a diverse T cell repertoire.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Timo
/
Microscopia de Fluorescência por Excitação Multifotônica
Limite:
Animals
Idioma:
En
Revista:
J Vis Exp
Ano de publicação:
2008
Tipo de documento:
Article