title: Confocal Imaging of Myeloid Cells in the Corneal Stroma of Live Mice.
abstract: The accessibility and transparency of the cornea makes it a good tissue model for monitoring immunological responses usingin vivoreal time imaging analysis (Leeet al., 2010; Tanet al., 2013). These corneal qualities have also allowed for high-resolutionin vivoimaging of non-ocular tissue transplanted into the anterior chamber of the mouse eye (Speieret al., 2008a; Speieret al., 2008b). This protocol was adapted from Speier (2008) to successfully assess real-timein vivomyeloid cell dynamics in wounded corneas ofc-fms-EGFP mice (Chanet al., 2013). Macrophage colony-stimulating factor (CSF-1) regulates the differentiation and proliferation of cells of the mononuclear phagocyte system. The activity of CSF-1 is mediated by the CSF-1 receptor that is encoded byc-fms(Csf1r) protooncogene. Thec-fmsgene is expressed in macrophage, trophoblast cell lineages, and to some extent granulocytes. In thec-fms-EGFP mice EGFP, enhanced green fluorescent protein, is driven under the Csf1r, colony stimulating factor 1 receptor, promoter and highlights myeloid cells (Sasmonoet al., 2003). This protocol can be further adapted to image other transgenic mice expressing fluorescent proteins.
authors.affiliation: Francis I. Proctor Foundation, University of California, San Francisco, California ; Department of Ophthalmology, University of California, San Francisco, California.
authors.forename: Matilda F
authors.initials: MF
authors.lastname: Chan
authors.affiliation: Department of Anatomy, University of California, San Francisco, USA.
authors.forename: Zena
authors.initials: Z
authors.lastname: Werb