Inducing Axonal Varicosities with a Micromechanical Stimulus on Neurons

In This Article

Abstract

Source: Servello, D., et al. A Microbiomechanical System for Studying Varicosity Formation and Recovery in Central Neuron Axons. J. Vis. Exp. (2018).

This video demonstrates the induction of axonal varicosities by applying micromechanical stimulus on neurons. The system comprises a micropipette attached to a syringe containing buffer via tubing. Using a micromanipulator, the micropipette is positioned above the neurons. Fluid pressure is then applied to induce varicosities in axons.

Protocol

1. Setting Up the Puffing Pipette Apparatus

NOTE: There are five basic components to this set-up: the glass pipette, the tubing, the syringe, the micromanipulator, and the buffer (Hank's) Table 1. Any buffer that has physiologically relevant salt concentrations could be used in this set-up.

Pull a borosilicate pipette to have around 45-µm-diameter opening tip using a micropipette puller and the specifications in Table 2.
NOTE: It was found that 45 µm is a suitable size for the diameter of the pipette opening under experimental conditions. Slight....

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Results

Microscope setup for cell observation; includes micromanipulator, neuron cultures, and focal plane diagram.
Figure 1: Schematic and photograph of puffing apparatus. (A) Photographs showing the overall microscope set-up. (B) The puffing apparatus showing the glass pipette held by the micromanipulator and connected to the rubber tubing. (C) Phase contras.......