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.

[1]  Carol A Barnes,et al.  Experience-Dependent Coincident Expression of the Effector Immediate-Early Genes Arc and Homer 1a in Hippocampal and Neocortical Neuronal Networks , 2002, The Journal of Neuroscience.

[2]  Margaret Werner-Washburne,et al.  Hyperspectral microarray scanning: impact on the accuracy and reliability of gene expression data , 2005, BMC Genomics.

[3]  Jon R. Schoonover,et al.  Multivariate Curve Resolution in the Analysis of Vibrational Spectroscopy Data Files , 2003, Applied spectroscopy.

[4]  J. D. McGaugh,et al.  Experience-Dependent Gene Expression in the Rat Hippocampus after Spatial Learning: A Comparison of the Immediate-Early GenesArc, c-fos, and zif268 , 2001, The Journal of Neuroscience.

[5]  M E Dickinson,et al.  Multi-spectral imaging and linear unmixing add a whole new dimension to laser scanning fluorescence microscopy. , 2001, BioTechniques.

[6]  C. Barnes,et al.  Spatial exploration induces ARC, a plasticity‐related immediate‐early gene, only in calcium/calmodulin‐dependent protein kinase II‐positive principal excitatory and inhibitory neurons of the rat forebrain , 2006, The Journal of comparative neurology.

[7]  D. Haaland,et al.  Design, construction, characterization, and application of a hyperspectral microarray scanner. , 2004, Applied optics.

[8]  Thomas D. Nielsen,et al.  Hyperspectral imaging: a novel approach for microscopic analysis. , 2001, Cytometry.

[9]  Bruce L. McNaughton,et al.  Environment-specific expression of the immediate-early gene Arc in hippocampal neuronal ensembles , 1999, Nature Neuroscience.

[10]  P. Kotula,et al.  Automated Analysis of SEM X-Ray Spectral Images: A Powerful New Microanalysis Tool , 2003, Microscopy and Microanalysis.

[11]  David M. Haaland,et al.  Multivariate curve resolution for hyperspectral image analysis: applications to microarray technology , 2003, SPIE BiOS.

[12]  M.L. Huebschman,et al.  Characteristics and capabilities of the hyperspectral imaging microscope , 2002, IEEE Engineering in Medicine and Biology Magazine.

[13]  R. Pepperkok,et al.  Spectral imaging and its applications in live cell microscopy , 2003, FEBS letters.