Advanced imaging of multiple mRNAs in brain tissue using a custom hyperspectral imager and multivariate curve resolution.
Simultaneous imaging of multiple cellular components is of tremendous importance in the study of complex biological systems, but the inability to use probes with similar emission spectra and the time consuming nature of collecting images on a confocal microscope are prohibitive. Hyperspectral imaging technology, originally developed for remote sensing applications, has been adapted to measure multiple genes in complex biological tissues. A spectral imaging microscope was used to acquire overlapping fluorescence emissions from specific mRNAs in brain tissue by scanning the samples using a single fluorescence excitation wavelength. The underlying component spectra obtained from the samples are then separated into their respective spectral signatures using multivariate analyses, enabling the simultaneous quantitative measurement of multiple genes either at regional or cellular levels.
Sutherland
Vicki L
VL
Arizona Research Laboratories, Division of Neural Systems Memory & Aging, University of Arizona, Tucson, AZ, United States.
Timlin
Jerilyn A
JA
Nieman
Linda T
LT
Guzowski
John F
JF
Chawla
Monica K
MK
Worley
Paul F
PF
Roysam
Badri
B
McNaughton
Bruce L
BL
Sinclair
Michael B
MB
Barnes
Carol A
CA
D000818:Animals; D001921:Brain / Q000378:metabolism*; D019044:Image Cytometry; D008297:Male; D036641:Microscopy, Fluorescence, Multiphoton; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger