Department of Anesthesiology, Renji Hospital, Shanghai Jiaotong University, School of Medicine; Key Laboratory of Anesthesiology (Shanghai Jiaotong University), Ministry of Education; Department of Anesthesiology, Shanghai General Hospital, Shanghai Jiaotong University.
Department of Anesthesiology, Renji Hospital, Shanghai Jiaotong University, School of Medicine; Key Laboratory of Anesthesiology (Shanghai Jiaotong University), Ministry of Education.
J Vis Exp. 2024 Jul 19(209). doi: 10.3791/66262.
Stereotaxic injection of a specific brain region constitutes a fundamental experimental technique in basic neuroscience. Researchers commonly base their choice of stereotaxic injection parameters on mouse brain atlases or published materials that employed various populations/ages of mice and different stereotaxic equipment, necessitating further validation of the stereotaxic coordinate parameters. The efficacy of calcium imaging, chemogenetic, and optogenetic manipulations relies on the precise expression of reporter genes within the region of interest, often requiring several weeks of effort. Thus, it is a time-consuming task if the coordinates of the target brain region are not verified in advance. Using an appropriate dye instead of a virus and implementing cryosectioning, researchers can observe the injection site immediately following dye administration. This facilitates timely adjustments to coordinate parameters in cases where discrepancies exist between the actual injection site and the theoretical position. Such adjustments significantly enhance the accuracy of viral expression within the target region in subsequent experiments.
立体定位注射特定脑区是基础神经科学中一项基本的实验技术。研究人员通常根据小鼠脑图谱或已发表的文献资料来选择立体定位注射参数，这些文献资料使用了不同群体/年龄的小鼠和不同的立体定位设备，因此需要进一步验证立体定位坐标参数。钙成像、化学遗传学和光遗传学操作的效果依赖于报告基因在感兴趣区域内的精确表达，这通常需要数周的努力。因此，如果目标脑区的坐标没有事先验证，那么这将是一项耗时的任务。使用适当的染料代替病毒，并实施冷冻切片，研究人员可以在染料给药后立即观察注射部位。如果实际注射部位与理论位置存在差异，这有助于及时调整坐标参数。这种调整显著提高了后续实验中目标区域内病毒表达的准确性。
