Optical imaging of intrinsic signal is a powerful technique for studying the functional organization of the brain [T. Bonhoeffer, D.S. Kim, D. Malonek, D. Shoham, A. Grinvald, Optical imaging of the layout of functional domains in area 17 and across the area 17/18 border in cat visual cortex, Eur. J. Neurosci. 7 (1995) 1973–1988; M. Hubener, D. Shoham, A. Grinvald, T. Bonhoeffer, Spatial relationships among three columnar systems in cat area 17, J. Neurosci. 17 (1997) 9270–9284; D. Malonek, A. Grinvald, Interactions between electrical activity and cortical microcirculation revealed by imaging spectroscopy: implications for functional brain mapping, Science 272 (1996) 551–554; A. Shmuel, A. Grinvald, Functional organization for direction of motion and its relationship to orientation maps in cat area 18, J. Neurosci. 16 (1996) 6945–6964] , , and . Three components of intrinsic optical signal can be distinguished. Two of these components can be attributed either to changes in blood volume or to changes in oxygen consumption [R.D. Frostig, E.E. Lieke, D.Y. Ts'o, A. Grinvald, Cortical functional architecture and local coupling between neuronal activity and the microcirculation revealed by in vivo high resolution optical imaging of intrinsic signals, Proc. Natl. Acad. Sci. U. S. A. 87 (1990) 6082–6086] . The origin of the third component is not yet clear but the component seems to be based on scattered light [H.U. Dodt, G. D'Arcangelo, E. Pestel, W. Zieglgansberger, The spread of excitation in neocortical columns visualized with infrared-dark field videomicroscopy, NeuroReport 7 (1996) 1553–1558; K. Holthoff, O.W. Witte, Intrinsic optical signals in rat neocortical slices measured with near-infrared dark-field microscopy reveal changes in extracellular space, J. Neurosci. 16 (1996) 2740–2749; B.A. MacVicar, D. Hochman, Imaging of synaptically evoked intrinsic optical signals in hippocampal slices, J. Neurosci. 11 (1991) 1458–1469; L. Trachsel, H.U. Dodt, W. Zieglgansberger, The intrinsic optical signal evoked by chiasm stimulation in the rat suprachiasmatic nuclei exhibits GABAergic day–night variation, Eur. J. Neurosci. 8 (1996) 319–328] , , and . A spectral fitting method with three components is used for the analysis of intrinsic optical signal [M. Nemoto, Y. Nomura, C. Sato, M. Tamura, K. Houkin, I. Koyanagi, H. Abe, Analysis of optical signals evoked by peripheral nerve stimulation in rat somatosensory cortex: dynamic changes in hemoglobin concentration and oxygenation, J. Cereb. Blood Flow Metab. 19 (1999) 246–259] . In order to validate the analysis, we need the knowledge on contribution of signal resulted from hemoglobin to total intrinsic optical signal. The exchange transfusion with fluorocarbon has the advantage that can change the spectral contribution of hemoglobin [M. Ferrari, M.A. Williams, D.A. Wilson, N.V. Thakor, R.J. Traystman, D.F. Hanley, Cat brain cytochrome-c oxidase redox changes induced by hypoxia after blood–fluorocarbon exchange transfusion, Am. J. Physiol. 269 (1995) H417–H424; A.L. Sylvia, C.A. Piantadosi, O2 dependence of in vivo brain cytochrome redox responses and energy metabolism in bloodless rats, J. Cereb. Blood Flow Metab. 8 (1988) 163–172] and . Here we describe a new method of the reduction of hemoglobin signal from somatosensory evoked optical intrinsic signal in rat cortex by the combination of exchange transfusion with fluorocarbon and imaging system of thinned skull cranial window. The method allows for the study of the synaptically evoked changes in light scattering as well as fluorescence of calcium indicator or voltage-sensitive dye without absorption of hemoglobin.Themes: Cellular biologyTopics: Imaging techniques
"In imaging the surface of the brain, use of intrinsic optical signals (IOS) and intrinsic optical imaging (IOI) to monitor neural activities in vivo becomes more recognized (Miller et al., 1993; Uchino et al., 1995; Ikeda et al., 1996; Kristal and Dubinsky, 1997; Lemasters et al., 1997; Johnson et al., 2000) and popularly utilized in the field of neurosciences (Aitken et al., 1999; Andrew et al., 1999; Nomura et al., 2000; Haller et al., 2001). But both IOS and IOI are highly affected by light scattering and absorption of the measured neural tissue or the brain due to morphological structures and hemodynamic aspects of the brain, respectively. "
[Show abstract][Hide abstract] ABSTRACT: In this study, we investigated dynamic changes in light scattering and hemoglobin oxygen saturation (S(sc)O(2)) on the rat spinal cord due to peripheral electrical stimulation by measuring near infrared (NIR) and visible spectroscopy, respectively. The spectral slope in the wavelength region between 700 and 900 nm is used as an index (S(NIR)) to quantify light scattering. With a 100-mum (source-detector separation) fiber-optic needle probe, optical reflectance was measured from the left lumbar region, specifically LL5, of the spinal cord surface at a height of 575 mum from the spinal cord surface. Graded electrical stimulations from 20 to 50 V, in increments of 10 V, were given to the plantar surface of the rat left hind paw for a period of 20 s. Changes in both light scattering (S(NIR)) and S(sc)O(2) were determined as a difference between the baseline and the maximum of slope value and hemoglobin oxygen saturation, respectively, during the stimulation period. There were significant differences in both S(NIR) and S(sc)O(2) during stimulation, with the average percentage changes of 10.9% and 15.5%, respectively. We observed that both S(NIR) and S(sc)O(2) measured at the spinal cord are insensitive to the intensity of the electrical stimulus, which is possibly caused by the nonlinear process of neurovascular coupling. Our finding essentially indicates that peripheral electrical stimulation results in significant changes in both light scattering and hemoglobin oxygen saturation on the rat spinal cord, and ignoring light scattering changes could lead to possible negative offsets of hemodynamic parameters (oxy-, deoxy-, and total hemoglobin concentrations) obtained in the functional optical imaging in the brain.
"Optical changes within smooth muscle can also occur in the absence of blood and may contribute to some of the slower scattering changes seen in hippocampal slices , . At least one study did not observe optical changes in vivo in the absence of blood, but neither dark-field nor birefringence illumination was used . A survey of birefringence responses from different nerves of crayfish and lobster showed that the temporal structure of the optical response depended on the type of nerve used and sufficient sensitivity to record from nerves as small as crayfish ventral cord (250 µm) . "
"entation domains ( see Fig . 9B ) . We suggest that this remaining component is also due to CBV changes in parenchymal microvessels rather than changes in Ls due to cellular swelling ( Holthoff and Witte , 1996 ; MacVicar and Hochman , 1991 ) because the activity - dependent Ls component is unlikely to significantly contribute to the OIS in vivo ( Nomura et al . , 2000 ) . Since , unlike pial vessels , parenchymal vessels may not be fully dilated at low BP , they can still respond to stimulus ( for CO 2 stimulation , see Gregory et al . , 1981 ) , suggesting CBV regulation within paren - chymal microvessels independently of parent artery ."
[Show abstract][Hide abstract] ABSTRACT: Since changes in oxygen consumption induced by active neurons are specific to cortical columns, the small and transient "dip" of deoxyhemoglobin signal, which indicates an increase in oxygen consumption, has been of great interest. In this study, we succeeded in enhancing and sustaining the dip in the deoxyhemoglobin-weighted 620-nm intrinsic optical imaging signals from a 10-s orientation-selective stimulation in cat visual cortex by reducing arterial blood pressure with sodium nitroprusside (a vasodilator) to mitigate the contribution of stimulus-induced blood supply. During this condition, intact spiking activity and a significant reduction of stimulus-induced blood volume changes (570-nm intrinsic signals) were confirmed. The deoxyhemoglobin signal from the prolonged dip was highly localized to iso-orientation domains only during the initial approximately 2 s; the signal specificity weakened over time although the domains were still resolvable after 2 s. The most plausible explanation for this time-dependent spatial specificity is that deoxyhemoglobin induced by oxygen consumption drains from active sites, where spiking activity occurs, to spatially non-specific downstream vessels over time. Our results suggest that the draining effect of pial and intracortical veins in dHb-based imaging techniques, such as blood oxygenation-level dependent (BOLD) functional MRI, is intrinsically unavoidable and reduces its spatial specificity of dHb signal regardless of whether the stimulus-induced blood supply is spatially specific.
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