Preparation of Neuron Balls as Hanging Drop Culture
3:24
Placing Neuron Balls on PLL-Coated Glass Coverslips and Culture Maintenance
4:29
Applying LRRTM2 Beads on Neuron Ball Culture With or Without Cell Bodies
6:07
Fluorescence Microscopy
7:19
Results: Analysis of Accumulation of Presynaptic Proteins in LRRTM2-Induced Presynapses of Axons of Neuron Ball Culture
9:15
Conclusion
副本
This protocol is suitable to investigate where presynaptic proteins originate:cell bodies or axons. And how presynaptic proteins accumulate in presynapses in an organized manner during the synapse formation. This technique can induce thousands of
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Presynapse formation is a dynamic process including accumulation of synaptic proteins in proper order. In this method, presynapse formation is triggered by beads conjugated with a presynapse organizer protein on axonal sheets of “neuron ball” culture, so that accumulation of synaptic proteins is easy to be analyzed during presynapse formation.