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Publication : Longitudinal in vivo two-photon fluorescence imaging.

First Author  Crowe SE Year  2014
Journal  J Comp Neurol Volume  522
Issue  8 Pages  1708-27
PubMed ID  24214350 Mgi Jnum  J:351723
Mgi Id  MGI:7663091 Doi  10.1002/cne.23502
Citation  Crowe SE, et al. (2014) Longitudinal in vivo two-photon fluorescence imaging. J Comp Neurol 522(8):1708-27
abstractText  Fluorescence microscopy is an essential technique for the basic sciences, especially biomedical research. Since the invention of laser scanning confocal microscopy in the 1980s, which enabled imaging both fixed and living biological tissue with 3D precision, high-resolution fluorescence imaging has revolutionized biological research. Confocal microscopy, by its very nature, has one fundamental limitation. Due to the confocal pinhole, deep tissue fluorescence imaging is not practical. In contrast (no pun intended), two-photon fluorescence microscopy allows, in principle, the collection of all emitted photons from fluorophores in the imaged voxel, dramatically extending our ability to see deep into living tissue. Since the development of transgenic mice with genetically encoded fluorescent protein in neocortical cells in 2000, two-photon imaging has enabled the dynamics of individual synapses to be followed for up to 2 years. Since the initial landmark contributions to this field in 2002, the technique has been used to understand how neuronal structure are changed by experience, learning, and memory and various diseases. Here we provide a basic summary of the crucial elements that are required for such studies, and discuss many applications of longitudinal two-photon fluorescence microscopy that have appeared since 2002.
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