Optical projection tomography as a tool for 3D microscopy and gene expression studies.
Current techniques for three-dimensional (3D) optical microscopy (deconvolution, confocal microscopy, and optical coherence tomography) generate 3D data by "optically sectioning" the specimen. This places severe constraints on the maximum thickness of a specimen that can be imaged. We have developed a microscopy technique that uses optical projection tomography (OPT) to produce high-resolution 3D images of both fluorescent and nonfluorescent biological specimens with a thickness of up to 15 millimeters. OPT microscopy allows the rapid mapping of the tissue distribution of RNA and protein expression in intact embryos or organ systems and can therefore be instrumental in studies of developmental biology or gene function.
Sharpe
James
J
Medical Research Council, Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK. james.sharpe@hgu.mrc.ac.uk
Ahlgren
Ulf
U
Perry
Paul
P
Hill
Bill
B
Ross
Allyson
A
Hecksher-Sørensen
Jacob
J
Baldock
Richard
R
Davidson
Duncan
D
D019412:Anatomy, Cross-Sectional; D000818:Animals; D004268:DNA-Binding Proteins / Q000032:analysis; D004622:Embryo, Mammalian / Q000033:anatomy & histology* / Q000378:metabolism*; D005314:Embryonic and Fetal Development; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D051542:Hepatocyte Nuclear Factor 3-beta; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008279:Magnetic Resonance Imaging; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D009154:Mutation; D009420:Nervous System / Q000196:embryology; D016900:Neurofilament Proteins / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D012984:Software; D013270:Stomach / Q000196:embryology; D014054:Tomography; D014157:Transcription Factors
D004268:DNA-Binding Proteins; C496503:Foxa2 protein, mouse; D016900:Neurofilament Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D051542:Hepatocyte Nuclear Factor 3-beta