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Research Papers: Imaging

Two-photon microscopy of cortical NADH fluorescence intensity changes: correcting contamination from the hemodynamic response

[+] Author Affiliations
Edward Baraghis

Harvard Medical School, Massachusetts General Hospital, Department of Radiology, 149 13th Street, Charlestown, Massachusetts 02129

École Polytechnique de Montréal, Département de génie électrique, Montreal, Canada H3C 3A7

Anna Devor

Harvard Medical School, Massachusetts General Hospital, Department of Radiology, 149 13th Street, Charlestown, Massachusetts 02129

University of California, Departments of Neurosciences and Radiology, San Diego, California 92093

Qianqian Fang, Vivek J. Srinivasan, Weicheng Wu, David A. Boas, Sava Sakadžić

Harvard Medical School, Massachusetts General Hospital, Department of Radiology, 149 13th Street, Charlestown, Massachusetts 02129

Frédéric Lesage

École Polytechnique de Montréal, Département de génie électrique, Montreal, Canada H3C 3A7

Cenk Ayata

Harvard Medical School, Massachusetts General Hospital, Department of Radiology, Neurovascular Research Laboratory, Charlestown, Massachusetts 02129

Massachusetts General Hospital, Department of Neurology, Stroke Service and Neuroscience Intensive Care Unit, Boston, Massachusetts 02114

Karl A. Kasischke

University of Rochester Medical Center, Department of Neurology, Center for Neural Development and Disease, Rochester, New York 14642

J. Biomed. Opt. 16(10), 106003 (October 03, 2011). doi:10.1117/1.3633339
History: Received April 20, 2011; Revised July 20, 2011; Accepted August 12, 2011; Published October 03, 2011; Online October 03, 2011
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Quantification of nicotinamide adenine dinucleotide (NADH) changes during functional brain activation and pathological conditions provides critical insight into brain metabolism. Of the different imaging modalities, two-photon laser scanning microscopy (TPLSM) is becoming an important tool for cellular-resolution measurements of NADH changes associated with cellular metabolic changes. However, NADH fluorescence emission is strongly absorbed by hemoglobin. As a result, in vivo measurements are significantly affected by the hemodynamics associated with physiological and pathophysiological manipulations. We model NADH fluorescence excitation and emission in TPLSM imaging based on precise maps of cerebral microvasculature. The effects of hemoglobin optical absorption and optical scattering from red blood cells, changes in blood volume and hemoglobin oxygen saturation, vessel size, and location with respect to imaging location are explored. A simple technique for correcting the measured NADH fluorescence intensity changes is provided, with the utilization of a parallel measurement of a physiologically inert fluorophore. The model is applied to TPLSM measurements of NADH fluorescence intensity changes in rat somatosensory cortex during mild hypoxia and hyperoxia. The general approach of the correction algorithm can be extended to other TPLSM measurements, where changes in the optical properties of the tissue confound physiological measurements, such as the detection of calcium dynamics.

Figures in this Article
© 2011 Society of Photo-Optical Instrumentation Engineers (SPIE)

Citation

Edward Baraghis ; Anna Devor ; Qianqian Fang ; Vivek J. Srinivasan ; Weicheng Wu, et al.
"Two-photon microscopy of cortical NADH fluorescence intensity changes: correcting contamination from the hemodynamic response", J. Biomed. Opt. 16(10), 106003 (October 03, 2011). ; http://dx.doi.org/10.1117/1.3633339


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