Quantitative Determination of Glymphatic Flow Using Spectrophotofluorometry
Quantitative Determination of Glymphatic Flow Using Spectrophotofluorometry
复制标题
使用分光光度荧光法定量测定类淋巴流量
DOI：
10.1007/s12264-020-00548-w
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发表时间：
2020-07
影响因子：
5.6
通讯作者：
Wang Linhui
中科院分区：
文献类型：
--
作者：
Zhang Yu;Song Jian;He Xuzhong;Xiong Jian;Xue Rong;Ge Jiahao;Lu Shiyu;Hu Die;Zhang Guoxing;Xu Guangyin;Wang Linhui
在鞘内注射荧光示踪剂之后，脑振动切片的离体成像已被广泛用于研究啮齿动物脑中的胶质淋巴系统。示踪剂进入大脑的渗透通常通过图像处理来量化，尽管这种方法需要大量的时间和手动操作。在这里，我们说明了一个简单的协议，使用分光荧光定量测定胶质淋巴活性。在脑池内或纹状体内注射荧光示踪剂后的特定时间点，收获某些脑区域和脊髓，并从组织中提取示踪剂。示踪剂的强度进行了分析，并从标准曲线定量其浓度。使用该方法，评估蛛网膜下CSF示踪剂向脑实质中的局部和动态递送，并且还确定示踪剂从脑中的清除。此外，使用我们的方法证实了老年小鼠脑中胶质淋巴流入的损伤。我们的方法在胶质淋巴活性的定量测定方面比成像方法更准确和有效，这将在临床前研究中有用。
Following intrathecal injection of fluorescent tracers, ex vivo imaging of brain vibratome slices has been widely used to study the glymphatic system in the rodent brain. Tracer penetration into the brain is usually quantified by image-processing, even though this approach requires much time and manual operation. Here, we illustrate a simple protocol for the quantitative determination of glymphatic activity using spectrophotofluorometry. At specific time-points following intracisternal or intrastriatal injection of fluorescent tracers, certain brain regions and the spinal cord were harvested and tracers were extracted from the tissue. The intensity of tracers was analyzed spectrophotometrically and their concentrations were quantified from standard curves. Using this approach, the regional and dynamic delivery of subarachnoid CSF tracers into the brain parenchyma was assessed, and the clearance of tracers from the brain was also determined. Furthermore, the impairment of glymphatic influx in the brains of old mice was confirmed using our approach. Our method is more accurate and efficient than the imaging approach in terms of the quantitative determination of glymphatic activity, and this will be useful in preclinical studies.
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影响因子：
4.6
作者：
Lundgaard I;Wang W;Eberhardt A;Vinitsky HS;Reeves BC;Peng S;Lou N;Hussain R;Nedergaard M
通讯作者：
Nedergaard M
DOI：
10.1126/science.1241224
发表时间：
2013-10-18
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Xie L;Kang H;Xu Q;Chen MJ;Liao Y;Thiyagarajan M;O'Donnell J;Christensen DJ;Nicholson C;Iliff JJ;Takano T;Deane R;Nedergaard M
通讯作者：
Nedergaard M
影响因子：
6.3
作者：
Lundgaard, Iben;Lu, Minh Lon;Nedergaard, Maiken
通讯作者：
Nedergaard, Maiken
影响因子：
3
作者：
SARIA, A;LUNDBERG, JM
通讯作者：
LUNDBERG, JM
影响因子：
3
作者：
Wolf MS;Chen Y;Simon DW;Alexander H;Ross M;Gibson GA;Manole MD;Bayır H;Kochanek PM;Clark RSB
通讯作者：
Clark RSB