Mangalath Aswathy, Raj Vishnu, Santhosh Rasitha, Thekkuveettil Anoopkumar
Division of Molecular Medicine, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum, Kerala- 695012, India.
Bio Protoc. 2023 Jun 20;13(12):e4697. doi: 10.21769/BioProtoc.4697.
A fascinating question in neuroscience is how sensory stimuli evoke calcium dynamics in neurons. is one of the most suitable models for optically recording high-throughput calcium spikes at single-cell resolution. However, calcium imaging in is challenging due to the difficulties associated with immobilizing the organism. Currently, methods for immobilizing worms include entrapment in a microfluidic channel, anesthesia, or adhesion to a glass slide. We have developed a new method to immobilize worms by trapping them in sodium alginate gel. The sodium alginate solution (5%), polymerized with divalent ions, effectively immobilizes worms in the gel. This technique is especially useful for imaging neuronal calcium dynamics during olfactory stimulation. The highly porous and transparent nature of alginate gel allows the optical recording of cellular calcium oscillations in neurons when briefly exposed to odor stimulation.
神经科学中一个引人入胜的问题是感觉刺激如何在神经元中引发钙动力学。[具体生物名称]是在单细胞分辨率下光学记录高通量钙峰的最合适模型之一。然而，由于固定该生物体存在困难，在[具体生物名称]中进行钙成像具有挑战性。目前，固定线虫的方法包括困在微流控通道中、麻醉或粘附在载玻片上。我们开发了一种通过将线虫捕获在海藻酸钠凝胶中来固定它们的新方法。与二价离子聚合的5%海藻酸钠溶液能有效地将线虫固定在凝胶中。这项技术对于在嗅觉刺激期间成像神经元钙动力学特别有用。海藻酸钠凝胶的高度多孔和透明性质使得在短暂暴露于气味刺激时能够光学记录神经元中的细胞钙振荡。
