Name: simultaneous evaluation of cerebral hemodynamics and light scattering properties of the in vivo rat brain using multispectral diffuse reflectance imaging
Uploaded: 2017-05-07T15:00:00
Description: Watch this Scientific Journal Video about Simultaneous Evaluation of Cerebral Hemodynamics and Light Scattering Properties of the In Vivo Rat Brain Using Multispectral Diffuse Reflectance Imaging at J

Part of this work was supported by a Grant-in-Aid for Scientific Research (C) from the Japanese Society for the Promotion of Science (25350520, 22500401, 15K06105) and the US-ARMY ITC-PAC Research and Development Project (FA5209-15-P-0175).

Animal care, preparation, and experimental protocols were approved by the Animal Research Committee of Tokyo University of Agriculture and Technology. For this methodology, the rat is housed in a controlled environment (24 &#176;C, 12 h light/dark cycle), with food and water available ad libitum.
1. Construction of a Conventional Multispectral Diffuse Reflectance Imaging System
<ol>
	<li>Mount nine narrowband optical interference filters with center wavelengths of 500, 520, 540, 560...

<ol>
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The most critical step in this protocol is the removal of the thinned skull region to make the cranial window; this should be performed carefully to avoid unexpected bleeding. This step is important for obtaining high-quality multispectral diffuse reflectance images with high accuracy. The use of a stereomicroscope is recommended for the surgical procedure if possible. Small pieces of gelatin sponge are useful for hemostasis.
The optical system described in this article passes a monochromatic ...

Multispectral diffuse reflectance imaging is the most common technique for obtaining a spatial map of intrinsic optical signals (IOSs) in cortical tissue. IOSs observed in the in vivo brain are mainly attributed to three phenomena: variations in light absorption and scattering properties due to cortical hemodynamics, variation in absorption depending on the reduction or oxidization of cytochromes in mitochondria, and variations in light scattering properties induced by morphological alterations1.
Light in the visible (VIS) to near-infrared (NIR) spectral range is effectively absorbed and scattered by biological tissue. The diffuse reflectance spectrum of the in vivo brain is characterized by absorption and scattering spectra. The reduced scattering coefficients &#956;s&#39; of brain tissue in the VIS-to-NIR wavelength range result in a monotonous scattering spectrum exhibiting smaller magnitudes at longer wavelengths. The reduced scattering coefficient spectrum &#956;s&#39;(&#955;) can be approximated to be in the form of the power law function2,3 as &#956;s&#39;(&#955;) = a &#215; &#955;-b. The scattering power b is related to the size of biological scatterers in living tissue2,3. Morphological alterations of the tissue and reduction of the viability of living cortical tissue can affect the size of the biological scatterers4,5,6,7,8,9.
An optical system for multispectral diffuse reflectance imaging can be easily constructed from an incandescent light source, simple optical components, and a monochromatic charged-coupled device (CCD). Therefore, various algorithms and optical systems for multispectral diffuse reflectance imaging have been used to evaluate cortical hemodynamics and/or tissue morphology10,11,12,13,14,15,16,17,18.
The method described in this article is used to visualize both the hemodynamics and light scattering properties of rat cerebral tissue in vivo using a conventional multispectral diffuse reflectance imaging system. The advantages of this method over alternative techniques are the ability to evaluate spatiotemporal changes in both cerebral hemodynamics and cortical tissue morphology, as well as its applicability to various brain dysfunction animal models. Therefore, the method will be appropriate for investigations of traumatic brain injury, epileptic seizure, stroke, and ischemia.

<table border="0" cellpadding="0" cellspacing="0" width="856">
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		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="36">
			<td height="36" style="height:36px;width:289px;">150-W halogen-lamp light source</td>
			<td style="width:281px;">Hayashi Watch Works Co., Ltd, Tokyo, Japan</td>
			<td style="width:119px;">LA-150SAE</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="36">
			<td height="36" style="height:36px;">Light guide</td>
			<td style="width:281px;">Hayashi Watch Works Co., Ltd, Tokyo, Japan</td>
			<td>LGC1-5L1000</td>
			<td></td>
		</tr>
		<tr height="36">
			<td height="36" style="height:36px;">Collecting lens</td>
			<td style="width:281px;">Hayashi Watch Works Co., Ltd, Tokyo, Japan</td>
			<td>SH-F16</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@500nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#65088</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@520nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#65093</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@540nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#65096</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@560nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#67766</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@570nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#67767</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@580nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#65646</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@600nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#65102</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@730nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#65115</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Interference filters l@760nm</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>#67777</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Motorized filter wheel&#160;</td>
			<td>Andover Corporation, NH, USA</td>
			<td>FW-MOT-12.5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">16-bit cooled CCD camera</td>
			<td>Bitran, Japan</td>
			<td>BS-40</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Video zoom lens</td>
			<td>Edmund Optics Japan Ltd, Tokyo, Japan</td>
			<td>VZMTM300i</td>
			<td></td>
		</tr>
		<tr height="36">
			<td height="36" style="height:36px;width:289px;">Spectralon white standard with 99% diffuse reflectance</td>
			<td>Labsphere Incorporated, North Sutton, NH, USA</td>
			<td>SRS-99-020</td>
			<td></td>
		</tr>
	</tbody>
</table>

simultaneous evaluation of cerebral hemodynamics and light scattering properties of the in vivo rat brain using multispectral diffuse reflectance imaging

The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional multispectral diffuse reflectance imaging system. This system is constructed from a broadband white light source, a motorized filter wheel with a set of narrowband interference filters, a light guide, a collecting lens, a video zoom lens, and a monochromatic charged-coupled device (CCD) camera. An ellipsoidal cranial window is made in the skull bone of a rat under isoflurane anesthesia to capture in vivo multispectral diffuse reflectance images of the cortical surface. Regulation of the fraction of inspired oxygen using a gas mixture device enables the induction of different respiratory states such as normoxia, hyperoxia, and anoxia. A Monte Carlo simulation-based multiple regression analysis for the measured multispectral diffuse reflectance images at nine wavelengths (500, 520, 540, 560, 570, 580, 600, 730, and 760 nm) is then performed to visualize the two-dimensional maps of hemodynamics and the light scattering properties of the in vivo rat brain.

Representative spectral images of diffuse reflectance acquired from in vivo rat brains are shown in Figure 3. The images at 500, 520, 540, 560, 570, and 580 nm clearly visualize a dense network of blood vessels in the cerebral cortex. The deterioration of contrast between blood vessels and the surrounding tissue observed in the images at 600, 730, and 760 nm reflects the lower absorption of light by hemoglobin at longer and NIR wavelengths.
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Watch this Scientific Journal Video about Simultaneous Evaluation of Cerebral Hemodynamics and Light Scattering Properties of the In Vivo Rat Brain Using Multispectral Diffuse Reflectance Imaging at J

Simultaneous Evaluation of Cerebral Hemodynamics and Light Scattering Properties of the In Vivo Rat Brain Using Multispectral Diffuse Reflectance Imaging

The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional multispectral diffuse reflectance imaging system.

Simultaneous Evaluation of Cerebral Hemodynamics and Light Scattering Properties of the In Vivo Rat Brain Using Multispectral Diffuse Reflectance Imaging

The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional ...

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