Visualization of single molecules of mRNA in situ.
作者： Andrea M. Femino , Kevin Fogarty , Lawrence M. Lifshitz , Walter Carrington , Robert H. Singer
DOI: 10.1016/S0076-6879(03)61015-3
关键词: Optical sectioning 、 In situ 、 Digital image 、 Deconvolution 、 Digital imaging 、 Fluorescence in situ hybridization 、 Pixel 、 Biological system 、 Computer science 、 Microscope 、 Nanotechnology
摘要: Publisher Summary The chapter discusses methods and concepts that facilitate the detection identification of single molecules mRNA in situ using fluorescence hybridization (FISH). FISH is a very widely used technique cell biology. sensitivity major concern when implementing technology. methodology employs stringent imaging requirements include carefully calibrated quantitative epifluorescence digital microscope, three-dimensional (3D) optical sectioning, constrained iterative deconvolution, 3D interactive analysis software. point sources representing one hybridized probe has provided additional information concerning nature process. pixel does not suffice as unit for image involving hybridization. detailed characterization biological now possible FISH. Unprecedented detail can be revealed from been interrogated with engineered, quantitatively accurate probes, followed by deconvolution.
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