该研究得到了国家自然科学基金（81960226,81760223）的资助;云南省自然科学基金（202001AS070045,202301AY070001-011）

该实验方案已获得昆明医科大学动物伦理与使用委员会（kmmu20220945）的批准。本研究使用Sprague-Dawley（SD）两性大鼠，300-350g。
1. 诱导SAH模型
<ol><li>使用2%异氟烷和100%氧气麻醉大鼠。通过提供异氟烷（1%-3%）的连续吸入麻醉来维持麻醉。使用立体定位装置固定大鼠的头部（参见 材料表）。</li>
	<li>按照以下步骤创建 SAH 模型。<ol><li>将显微注射针插入鞍上蓄水池。然后，使用注射泵将来自大鼠尾动脉（不含抗凝剂）的自体血液注入鞍上池（参见 材料表）。</li>
		<li>使用上述坐标将显微注射针植入右侧脑室：前囟，-0.8 mm;横向，1.4毫米;深度，4 mm9。对于 SAH 组，注射 0.2 mL 大鼠尾动脉血。对于假组，施用相同体积的等渗盐水（图1A）。</li>
	</ol></li>
</ol>2. 脑切片的制备和稳定
<ol><li>使用2%异氟烷吸入深度麻醉大鼠。斩首后，取出整个大脑并将它们浸入冰冷的人工脑脊液 （ACSF） 中。 注：使用具有以下成分的新鲜制备的 ACSF 溶液：134 mM NaCl、2.8 mM KCl、29 mM NaHCO3、1.1 mM NaH 2 PO 4、1.5 mM MgSO4、2.5 mM CaCl 2和 10.11 mM D-葡萄糖（参见材料表）。将 pH 值保持在 7.3 到 7.4 之间。</li>
	<li>使用振动切片机（参见材料表）在用5%CO2和95%O 2充气的冰冷ACSF中制备厚度为200μm的SD大鼠的急性脑切片（图2）。</li>
	<li>将 200 μm 厚的急性脑切片放入浸没在 ACSF 中的尼龙网的储存室中。用95%O 2和5%CO2平衡ACSF溶液，保持35-37°C的温度范围。 让脑切片稳定2小时，让它们有时间调整（图3）。</li>
</ol>3. 用 TO-PRO-3 标记急性脑切片中的周细胞
注：使用示踪剂 TO-PRO-310 荧光标记来自急性脑切片的周细胞。
<ol><li>将荧光染料TO-PRO-3加入ACSF中，达到1μM的终浓度。 将急性脑切片在室温下与含有TO-PRO-3的ACSF在黑暗环境中孵育20分钟。 注意： 请记住在处理 TO-PRO-3 时戴上乳胶手套以提供保护。</li>
	<li>将携带脑切片的尼龙网过滤器从装载室转移到 6 孔板冲洗室（参见 材料表）。让脑切片在冲洗室中停留10分钟（图4D）。 注意：此冲洗步骤终止染色过程，减少背景标记，并防止非特异性染色。</li>
	<li>使用与 5% CO 2 和 95% O2 的混合物平衡的 ACSF 溶液冲洗制剂共 15 分钟。该冲洗步骤可阻止染料吸收并最大限度地减少背景标记（图4C）。</li>
</ol>4.急性脑切片中脑微循环的非重要周细胞染色
<ol><li>在黑暗环境中，将预装有TO-PRO-3的脑切片与Alexa Fluor 488（FITC-ISOB4;5μg/ mL，参见 材料表）偶联的异凝集素B4在37°C孵育30分钟。孵育后，在ACSF中冲洗脑切片15分钟（图4E）。</li>
	<li>像往常一样，在用 5% CO 2 和 95% O2 充气的 ACSF 中孵育脑切片。在37°C下将浓度为37μM的碘化丙啶（PI）加入两种溶液中。 这将标记非重要的脑周细胞。将制剂在该ACSF溶液中在黑暗中孵育60分钟（图4F）。</li>
	<li>接下来，用ACSF溶液冲洗制剂15分钟，以阻止染料吸收并最大限度地减少背景标记。</li>
</ol>5. 急性脑切片中重要和非重要脑周细胞的成像
<ol><li>使用塑料巴斯德移液管轻轻地将脑切片转移到玻璃底共聚焦培养皿（见材料表）上。用先前用 5% CO 2 和 95% O2 平衡的 ACSF 溶液填充培养皿。使用铁网将大鼠脑切片固定到位。</li>
	<li>将玻璃底共聚焦培养皿转移到共聚焦显微镜的载物台上。将急性脑切片放置在培养皿底部，并在共聚焦显微镜成像期间用95%O 2和5%CO2的混合物平衡的ACSF溶液灌注9（图5A和图5Ab'）。</li>
	<li>使用 40 倍物镜可视化大脑皮质微血管系统的壁细胞。使用兼容软件捕获图像堆栈（参见 材料表）。对IB4（激发/发射460nm/520nm）、PI（激发/发射545nm/595nm）和TO-PRO-3（激发/发射606nm/666nm）使用适当的滤光片（图5D）。 注意：使用以下详细信息捕获图像：40×DIC N1 物镜，3 × 12 位：512 × 512 像素（0.22 × 0.22 mm），校准：0.42 μm/px。</li>
	<li>根据脑微血管系统和周细胞的网络和形态识别它们。在每个脑切片上找到一个包含脑微血管系统网络的特定区域并捕获图像。</li>
	<li>仔细记录成像位置，以确保后续捕获的一致性（图5B，C）。如需进一步处理和分析，请使用图像分析软件（ImageJ）和照片编辑软件（参见材料表）。</li>
</ol>

蛛网膜下腔出血后脑周细胞收缩力的可视化

<ol>
	<li>Dalkara, T., Gursoy-Ozdemir, Y., Yemisci, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+microvascular+pericytes+in+health+and+disease.">Brain microvascular pericytes in health and disease.</a> Acta Neuropathologica. 122 (1), 1-9 (2011).</li><li>Dore-Duffy, P., Cleary, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphology+and+properties+of+pericytes.">Morphology and properties of pericytes.</a> Methods in Molecular Biology (Clifton, N.J). 686, 49-68 (2011).</li><li>Peppiatt, C. M., Howarth, C., Mobbs, P., Attwell, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bidirectional+control+of+CNS+capillary+diameter+by+pericytes.">Bidirectional control of CNS capillary diameter by pericytes.</a> Nature. 443 (7112), 700-704 (2006).</li><li>Attwell, D., Mishra, A., Hall, C. N., O'Farrell, F. M., Dalkara, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+is+a+pericyte.">What is a pericyte.</a> Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism. 36 (2), 451-455 (2016).</li><li>Yemisci, M., Gursoy-Ozdemir, Y., Vural, A., Can, A., Topalkara, K., Dalkara, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pericyte+contraction+induced+by+oxidative-nitrative+stress+impairs+capillary+reflow+despite+successful+opening+of+an+occluded+cerebral+artery.">Pericyte contraction induced by oxidative-nitrative stress impairs capillary reflow despite successful opening of an occluded cerebral artery.</a> Nature Medicine. 15 (9), 1031-1037 (2009).</li><li>Korte, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noradrenaline+released+from+locus+coeruleus+axons+contracts+cerebral+capillary+pericytes+via+&#945;2+adrenergic+receptors.">Noradrenaline released from locus coeruleus axons contracts cerebral capillary pericytes via &#945;2 adrenergic receptors.</a> Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism. , (2023).</li><li>Hirunpattarasilp, C., Barkaway, A., Davis, H., Pfeiffer, T., Sethi, H., Attwell, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperoxia+evokes+pericyte-mediated+capillary+constriction.">Hyperoxia evokes pericyte-mediated capillary constriction.</a> Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism. 42 (11), 2032-2047 (2022).</li><li>Neuhaus, A. A., Couch, Y., Sutherland, B. A., Buchan, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+method+to+study+pericyte+contractility+and+responses+to+ischaemia+in+vitro+using+electrical+impedance.">Novel method to study pericyte contractility and responses to ischaemia in vitro using electrical impedance.</a> Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism. 37 (6), 2013-2024 (2017).</li><li>Gong, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+TRPM4+Activity+in+cerebral+artery+myocytes+contributes+to+cerebral+blood+flow+reduction+after+subarachnoid+hemorrhage+in+rats.">Increased TRPM4 Activity in cerebral artery myocytes contributes to cerebral blood flow reduction after subarachnoid hemorrhage in rats.</a> Neurotherapeutics: The Journal of the American Society for Experimental NeuroTherapeutics. 16 (3), 901-911 (2019).</li><li>Mai-Morente, S. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pericyte+mapping+in+cerebral+slices+with+the+far-red+fluorophore+TO-PRO-3.">Pericyte mapping in cerebral slices with the far-red fluorophore TO-PRO-3.</a> Bio-protocol. 11 (22), e4222 (2021).</li><li>Mai-Morente, S. P., Marset, V. M., Blanco, F., Isasi, E. E., Abudara, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+nuclear+fluorescent+dye+identifies+pericytes+at+the+neurovascular+unit.">A nuclear fluorescent dye identifies pericytes at the neurovascular unit.</a> Journal of Neurochemistry. 157 (4), 1377-1391 (2021).</li><li>Zhao, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rationale+for+the+real-time+and+dynamic+cell+death+assays+using+propidium+iodide.">Rationale for the real-time and dynamic cell death assays using propidium iodide.</a> Cytometry. Part A: The Journal of the International Society for Analytical Cytology. 77 (4), 399-405 (2010).</li><li>Van Hooijdonk, C. A., Glade, C. P., Van Erp, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TO-PRO-3+iodide:+A+novel+HeNe+laser-excitable+DNA+stain+as+an+alternative+for+propidium+iodide+in+multiparameter+flow+cytometry.">TO-PRO-3 iodide: A novel HeNe laser-excitable DNA stain as an alternative for propidium iodide in multiparameter flow cytometry.</a> Cytometry. 17 (2), 185-189 (1994).</li><li>Lacar, B., Herman, P., Platel, J. C., Kubera, C., Hyder, F., Bordey, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+progenitor+cells+regulate+capillary+blood+flow+in+the+postnatal+subventricular+zone.">Neural progenitor cells regulate capillary blood flow in the postnatal subventricular zone.</a> The Journal of Neuroscience: The Official Journal of the Society for Neuroscience. 32 (46), 16435-16448 (2012).</li><li>Mai-Morente, S. P., Marset, V. M., Blanco, F., Isasi, E. E., Abudara, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+nuclear+fluorescent+dye+identifies+pericytes+at+the+neurovascular+unit.">A nuclear fluorescent dye identifies pericytes at the neurovascular unit.</a> Journal of Neurochemistry. 157 (4), 1377-1391 (2021).</li><li>Hezel, M., Ebrahimi, F., Koch, M., Dehghani, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Propidium+iodide+staining:+a+new+application+in+fluorescence+microscopy+for+analysis+of+cytoarchitecture+in+adult+and+developing+rodent+brain.">Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain.</a> Micron (Oxford, England). 43 (10), 1031-1038 (2012).</li><li>Mathiisen, T. M., Lehre, K. P., Danbolt, N. C., Ottersen, O. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+perivascular+astroglial+sheath+provides+a+complete+covering+of+the+brain+microvessels:+An+electron+microscopic+3D+reconstruction.">The perivascular astroglial sheath provides a complete covering of the brain microvessels: An electron microscopic 3D reconstruction.</a> Glia. 58 (9), 1094-1103 (2010).</li></ol>

提交人声明，他们没有相互竞争的经济利益。

开发的是高分辨率共聚焦成像技术，用于可视化重要的脑周细胞、非重要的脑周细胞和脑切片中的微血管系统。在急性大鼠脑切片中，该过程需要用 TO-PRO-311 初始标记周细胞，然后用 IB412 标记微血管内皮细胞;随后，使用PI进行死亡周细胞的鉴定。该协议简单明了，可重复，并且高度适用于功能研究。
为了特异性地追踪神经系统内的脑周细?...

脑周细胞以其细长的突起和突出的细胞体为特征，环绕着微循环 1,2。虽然脑血流增加主要由毛细血管扩张驱动，但较小的动脉表现出较慢的扩张速度3。周细胞收缩力对毛细血管直径和周细胞形态产生影响，影响血管动力学4.脑周细胞的收缩导致毛细血管收缩，在病理情况下，过度收缩可能会阻碍红细胞流动5.多种因素，包括从蓝斑释放的去甲肾上腺素，可诱导毛细血管内的脑周细胞收缩6.周细胞在脑血流中起调节作用，表现出 20-HETE 合成，在高氧期间充当氧传感器7。氧化-硝化应激引发的脑周细胞收缩对毛细血管产生不利影响5.尽管对脑周细胞收缩进行了体内和体外研究8，但关于脑切片中活和非活脑周细胞成像的知识仍然有限。
至关重要的是，脑周细胞的组织固定后成像会影响其活力和随后的收缩性评估。此外，在神经系统疾病（例如蛛网膜下腔出血 - SAH）等情况下，脑周细胞的转基因标记无法区分活的和无活的周细胞，正如我们初步的 SAH 诱导的脑周细胞死亡研究所证实的那样9. 
为了克服这些挑战，我们使用 TO-PRO-3 标记活周细胞，同时用碘化丙啶 （PI） 染色死亡周细胞。我们使用高分辨率共聚焦成像技术来可视化脑切片中活的和无活的脑周细胞，同时在成像过程中保持切片活性。本文旨在提出一种可重复的方法，用于对脑切片中活的和无活的脑周细胞进行成像，作为探究脑周细胞对 SAH 后脑微循环影响的宝贵工具。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>6-well plate</td>
			<td>ABC biochemistry</td>
			<td>ABC703006</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Adobe Photoshop</td>
			<td>Adobe</td>
			<td>Adobe Illustrator CS6 16.0.0</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Aluminium foil</td>
			<td>MIAOJIE</td>
			<td>225 mm x 273 mm</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>CaCl2&#183;2H2O</td>
			<td>Sigma-Aldrich</td>
			<td>C3881</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Confocal imaging software</td>
			<td>Nikon</td>
			<td>NIS-Elements 4.10.00</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Confocal Laser Scanning Microscope</td>
			<td>Nikon</td>
			<td>N-SIM/C2si</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Gas tank (5% CO2, 95% O2)</td>
			<td>PENGYIDA</td>
			<td>40L</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Glass Bottom Confocal Dishes</td>
			<td>Beyotime</td>
			<td>FCFC020-10pcs</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>Sigma-Aldrich</td>
			<td>G5767</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Glue</td>
			<td>EVOBOND</td>
			<td>KH-502</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Ice machine</td>
			<td>XUEKE</td>
			<td>IMS-20</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Image analysis software</td>
			<td>National Institutes of Health</td>
			<td>Image J</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Inhalation anesthesia system</td>
			<td>SCIENCE</td>
			<td>QAF700</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Isolectin B 4-FITC</td>
			<td>SIGMA</td>
			<td>L2895&#8211;2MG</td>
			<td>Store aliquots at &#8211;20 &#176;C</td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma-Aldrich</td>
			<td>7447&#8211;40&#8211;7</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>KH2PO4</td>
			<td>Sigma-Aldrich</td>
			<td>P0662</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>MgSO4</td>
			<td>Sigma-Aldrich</td>
			<td>M7506</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma-Aldrich</td>
			<td>7647&#8211;14&#8211;5</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>NaH2PO4&#183;H2O</td>
			<td>Sigma-Aldrich</td>
			<td>10049&#8211;21&#8211;5</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>NaHCO3</td>
			<td>Sigma-Aldrich</td>
			<td>S5761</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Pasteur pipette</td>
			<td>NEST Biotechnology</td>
			<td>318314</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Peristaltic Pump</td>
			<td>Scientific Industries Inc</td>
			<td>Model 203</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Propidium (Iodide)</td>
			<td>Med Chem Express</td>
			<td>HY-D0815/CS-7538</td>
			<td>Store aliquots at &#8211;20 &#176;C</td>
		</tr>
		<tr>
			<td>Stereotaxic apparatus</td>
			<td>SCIENCE</td>
			<td>QA</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Syringe pump</td>
			<td>Harvard PUMP</td>
			<td>PUMP 11 ELITE Nanomite</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Thermostatic water bath</td>
			<td>OLABO</td>
			<td>HH-2</td>
			<td>RT</td>
		</tr>
		<tr>
			<td>Vibrating microtome</td>
			<td>Leica</td>
			<td>VT1200</td>
			<td>RT</td>
		</tr>
	</tbody>
</table>

imaging vital and non-vital brain pericytes in brain slices following subarachnoid hemorrhage

周细胞是位于大脑微循环内的关键壁细胞，在通过收缩力调节主动调节脑血流方面发挥着关键作用。传统上，它们的收缩力是通过观察特定情况下的形态变化和附近的毛细管直径变化来衡量的。然而，组织固定后、评估活力和随之而来的成像脑周周细胞的周细胞收缩力会受到影响。同样，对脑周细胞进行基因标记在区分活细胞和非活周细胞方面存在不足，特别是在蛛网膜下腔出血（SAH）等神经系统疾病中，我们的初步研究证实了脑周细胞的死亡。已经设计了一种可靠的方案来克服这些限制，能够在脑切片中同时对功能性和非功能性脑周细胞进行荧光标记。这种标记方法允许高分辨率共聚焦显微镜可视化，同时标记脑切片微血管系统。这种创新方案提供了一种评估脑周细胞收缩性、其对毛细血管直径和周细胞结构的影响的方法。在 SAH 背景下研究脑周细胞收缩力可以深入了解其对脑微循环的影响。

Brain pericytes, distinguished by their slender protuberances and protruding cell bodies, encircle the microcirculation1,2. While cerebral blood flow augmentation is predominantly driven by capillary dilation, smaller arteries exhibit slower rates of dilation3. Pericyte contractility exerts influence over capillary diameter and pericyte morphology, impacting vascular dynamics4. Contraction of brain pericytes leads to capillary constriction, and in pathological scenarios, excessive contraction may impede erythrocyte flow5. Various factors, including norepinephrine released from the locus coeruleus, can induce brain pericyte contraction within capillaries6. With a regulatory role in cerebral blood flow, pericytes exhibit 20-HETE synthesis, serving as an oxygen sensor during hyperoxia7. Oxidative-nitrative stress-triggered contraction of brain pericytes detrimentally affects capillaries5. Despite both in vivo and ex vivo investigations into brain pericyte contraction8, limited knowledge persists regarding the imaging of viable and non-viable brain pericytes within brain slices.
Crucially, post-tissue fixation imaging of brain pericytes compromises their vitality and subsequent contractility assessment. Moreover, in scenarios such as neurological disorders (e.g., subarachnoid hemorrhage - SAH), transgenic labeling of brain pericytes fails to differentiate between viable and non-viable pericytes, as confirmed by our preliminary SAH-induced brain pericyte death study9. 
To surmount these challenges, we employed TO-PRO-3 to label live pericytes, while deceased ones were stained with propidium iodide (PI). We used high-resolution confocal imaging technologies to visualize viable and non-viable brain pericytes in brain slices while preserving slice activity during imaging. This article aims to present a reproducible method for imaging viable and non-viable brain pericytes in brain slices, serving as a valuable tool to probe the impact of brain pericytes on cerebral microcirculation post SAH.

作者聚焦：啮齿动物蛛网膜下腔出血后周细胞成像 

蛛网膜下腔出血后脑切片中重要和非重要脑周细胞的成像

Our study showed a significant increase in pericytes deaths. So starting at a sick horse after SAH, it demonstrated a positive correlation between the number of low vital parasites and the time elapsed after SAH. Imaging vital and the non-vital brain parasites in brain slices following SAH and the parasite isolation techniques, have advanced the research in my field.Currently, it's quite difficult to fully distinguish between dead parasites and dead endothelial cells. Our study find contraction-dependent apoptosis in parasites after SAH. We have developed a very reliable protocol, which enables the fluorescent labeling, both functional and a non-functional brain parasite in brain sections, and the same time.

在正常生理条件下，脑周细胞一般不会发生细胞死亡。 图 6 说明了这种现象，黄色表示存在重要的脑周细胞;脑周细胞未显示 PI 染色，表明其活力。为了进一步研究细胞死亡后周细胞是否仍然附着在微血管系统上，在SAH大鼠模型中采用了方法，并进行了随后的成像。
已经开发了对 SAH 后脑切片中重要和非重要脑周细胞进行成像的方法。如 <strong class="xfig"...

Watch this Scientific Journal Video about Imaging Pericytes Post-Subarachnoid Hemorrhaging in Rodents at JoVE.com

初步调查证实，蛛网膜下腔出血 （SAH） 会导致脑周细胞死亡。评估 SAH 后周细胞收缩力需要区分活的和无活的脑周细胞。因此，已经开发了一种在脑切片中同时标记活和无活脑周细胞的程序，便于使用高分辨率共聚焦显微镜进行观察。

Pericytes are crucial mural cells situated within cerebral microcirculation, pivotal in actively modulating cerebral blood flow via contractility ...

我们的研究表明周细胞死亡显着增加。因此，从 SAH 后的病马开始，它证明了低生命寄生虫的数量与 SAH 后经过的时间之间存在正相关关系。在SAH和寄生虫分离技术之后，对脑切片中的重要和非重要脑寄生虫进行成像，推动了我所在领域的研究。目前，很难完全区分死去的寄生虫和死去的内皮细胞。我们的研究发现 SAH 后寄生虫的收缩依赖性细胞凋亡。我们已经开发了一种非常可靠的方案，可以同时在脑切片中对功能性和非功能性脑寄生虫进行荧光标记。

imaging-vital-non-vital-brain-pericytes-brain-slices-following

Research

JoVE Journal

Neuroscience

Imaging Vital and Non-vital Brain Pericytes in Brain Slices following Subarachnoid Hemorrhage

701 次观看.  The First Affiliated Hospital of Kunming Medical University. 我们的研究表明周细胞死亡显着增加。因此，从 SAH 后的病马开始，它证明了低生命寄生虫的数量与 SAH 后经过的时间之间存在正相关关系。在SAH和寄生虫分离技术之后，对脑切片中的重要和非重要脑寄生虫进行成像，推动了我所在领域的研究。目前，很难完全区分死去的寄生虫和死去的内皮细胞。我们的研究发现 SAH 后寄生虫的收缩依赖性细胞凋亡。我们已经开发了一?...