Abstract
This chapter demonstrates the use of two-photon excitation microscopy to monitor cellular metabolism in ocular tissue. Light can be used as a noninvasive probe to monitor cellular function in cells, tissues and organs. Cellular metabolism can be noninvasively optically monitored by the technique of redox fluorometry.1 This optical technique has been applied to the eye2–4 and quantitative analysis of the various pyridine nucleotides have been measured in individual cellular layers of the normoxic and anoxic eyes. The noninvasive optical probes are the intrinsic fluorescent pyridine nucleotides, and the flavoproteins. The pyridine nucleotides, nicotinamide adenine dinucleotide, NAD(P)H are excited in the region 365 nm and fluoresce in the region 400–500 nm. The flavoproteins are excited in the region 450 nm and fluoresce in the region 500–600 nm. Typically the fluorescence is imaged and changes in fluorescence intensity follow changes in cell oxidative metabolism. Optical sections of micron thickness can be combined to visualize the full three-dimensional image of thick tissues.
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References
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Masters, B.R. (1996). Three-Dimensional Optical Functional Imaging of Tissue with Two-Photon Excitation Laser Scanning Microscopy. In: Kohen, E., Hirschberg, J.G. (eds) Analytical Use of Fluorescent Probes in Oncology. NATO ASI Series, vol 286. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5845-3_20
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DOI: https://doi.org/10.1007/978-1-4615-5845-3_20
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