A method of marking neurons on brain slices using patch clamp technique

doi:10.12047/j.cjap.6035.2021.031

Abstract

Abstract: Objective: To introduce a method of marking neurons using patch clamp technique. Methods: The brain slices of the target area was cut with a vibrating microtome. The glass microelectrode was perfused with the electrode liquid containing Neurobiotin^TM Tracer, and the whole-cell patch-clamp recording was performed. After recording, the brain slices were fixed and rinsed with 4% paraformaldehyde. After stained in phosphate buffer with Streptavidin-Texas Red and Triton X-100 for at least 2 hours, the neurons can be observed under a fluorescence microscope. Results: The cell membrane voltage was clamped at -70 mV, and the neuron showed a gradually increasing membrane current after step stimulation. When recording in the current clamp mode, the step stimulus caused the neuron to depolarize to the threshold potential and then burst into action potentials. The morphology of intact neurons with clear cell body and protrusions of a neuron could be observed under a fluorescence microscope. Conclusion: This method is suitable for observing the morphological features of the recorded neuron after patch clamp experiments, which is easy to operate, and the image is intuitive and clear.

Key words: whole-cell patch clamp, brain slices, neuronal morphology, fluorescence tags

CLC Number:

R338.8

GAO Jie, HUANG Yan-yun, ZHANG Yu-tong, DU Xiang-xin, ZHANG Li-na, HAO Na, GUO Xia, LI Jian-guo, ZHANG Yu. A method of marking neurons on brain slices using patch clamp technique[J]. CJAP, 2021, 37(4): 445-448.

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