| Cell Cycle Analysis in the Vertebrate Brain Using Immunolabeled Fresh Cell Nuclei
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Author:
Date:
2013-11-20
[Abstract] Flow cytometry, a standard technique used for quantitative analysis of isolated cells, is routinely employed by immunologists and oncologists to study DNA content, protein expression, and other functional parameters in blood and tumor cells. Unfortunately, the use of this technique by neurobiologists has been hampered by the complexity of the nervous system, whose constituting cells can hardly be dissociated to obtain samples of sufficient quality. We have developed a simplified and quick method to purify and immunolabel cell nuclei with high sensitivity and low background. Our protocol allows the discrimination of single nuclei from doublets and larger aggregates, obtaining low coefficients of variation for cell cycle analysis with propidium iodide. In addition, due to the reduced sample ...
[摘要] Flow cytometry, a standard technique used for quantitative analysis of isolated cells, is routinely employed by immunologists and oncologists to study DNA content, protein expression, and other functional parameters in blood and tumor cells. Unfortunately, the use of this technique by neurobiologists has been hampered by the complexity of the nervous system, whose constituting cells can hardly be dissociated to obtain samples of sufficient quality. We have developed a simplified and quick method to purify and immunolabel cell nuclei with high sensitivity and low background. Our protocol ...
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| Neuronal Morphology Analysis
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Author:
Date:
2013-08-05
[Abstract] This protocol describes how to visualize neuronal morphology and how to determine neuronal complexity of immature and mature hippocampal neurons in the mouse in vivo including tissue preparation, staining of brain sections and confocal cell analysis.
[摘要] 该协议描述了如何可视化神经元形态和如何确定小鼠体内未成熟和成熟海马神经元的神经元复杂性，包括组织制备，脑切片染色和共聚焦细胞分析。
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| Evaluation of Caspase-1 Activation and IL-1β Production in A Kainic Acid Microdyalisis Brain Injury Model
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Author:
Date:
2013-04-20
[Abstract] Intracerebral infusion of kainic acid (KA) by a microdialysis probe induces a focal swelling in the brain-perfused area which promotes inflammation (Compan et al., 2012; Oprica et al., 2003). The microdialysis technique allows the local in vivo perfusion of KA and the simultaneous collection of inflammatory mediators, and other neuroactive substances, released in the injured brain. This protocol also allows the perfusion of different solutions in each cerebral hemisphere at the same time. By perfusing KA in isotonic solution of Krebs-Ringer Bicarbonate (KRB) (280-290 mOsm) in one hippocampus and KA in hypertonic KRB solution (1,400-1,500 mOsm) in the contralateral side, we can evaluate in vivo the efficiency of hypertonic solutions in preventing ...
[摘要] 通过微透析探针脑内输注红藻氨酸（KA）在脑灌注区域诱导局灶性肿胀，这促进炎症（Compan等人，2012; Oprica等人， 。，2003）。微透析技术允许局部的体内灌注KA和同时收集炎性介质和其他神经活性物质，释放在受伤的脑中。该协议还允许在每个大脑半球同时灌注不同的溶液。通过在对侧中在一个海马中的Krebs-Ringer碳酸氢盐（KRB）（280-290mOsm）的等渗溶液中灌注KA和在高渗KRB溶液（1,400-1,500mOsm）中的KA，我们可以在体内评价高渗溶液在KA输注后预防由肿胀引起的炎症的效率。一旦已经诱导炎症反应，就有可能通过微透析探针输入胱天蛋白酶-1的生物素化特异性抑制剂，从而允许检测响应于损伤的IL-1产生的脑区域和细胞（Oprica等人， et al。，2003）。
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