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Device and method for evaluating neuron-like differentiation degree of PC12 cell

A PC12 and neuron technology, applied in measurement devices, instruments, scanning probe microscopy, etc., can solve the problem of difficult scanning and imaging of PC12 cells, and achieve the effect of strong operability

Inactive Publication Date: 2012-02-15
CHINA NAT ACAD NANOTECH & ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for SICM under continuous negative feedback control mode, it is difficult to scan and image neuron-like PC12 cells with sharp vertical height changes at high resolution.

Method used

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  • Device and method for evaluating neuron-like differentiation degree of PC12 cell
  • Device and method for evaluating neuron-like differentiation degree of PC12 cell
  • Device and method for evaluating neuron-like differentiation degree of PC12 cell

Examples

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Embodiment

[0026] Embodiment: a kind of device for assessing the degree of neuron-like differentiation of PC12 cells (see Figure 4 ), which is characterized in that it is a non-contact, high-resolution probe-hopping mode scanning ion conductance microscope (HPICM) that detects the surface microstructure of living biological samples under physiological liquid culture conditions, and functionally NGF-induced differentiation A device consisting of two parts of the patch clamp system for evaluating the electrical excitability of PC12 cells; the device for evaluating the degree of neuron-like differentiation of PC12 cells includes a glass microprobe filled with electrolyte, a glass microprobe placed Ag / AgCl electrode immersed in the electrolyte, reference Ag / AgCl electrode, petri dish containing cells and cell culture medium, HPICM scanning control system, Z-direction HPICM scanning probe, XY-direction HPICM scanning table, patch clamp Amplifier, digital-to-analog / analog-to-digital converter...

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Abstract

The invention discloses a device for evaluating the neuron-like differentiation degree of a PC12 cell. The device consists of a hopping probe ion conductance microscope (HPICM) and a patch clamp system. An evaluating method comprises the following steps of: obtaining a glass microprobe for observing the micro-topography of the PC12 cell with an HPLCM technology; scanning and imaging the surface topographies of living neuron-like PC12 cell at different NGF (Nerve Growth Factor) induced differentiation time points with the HPLCM technology; analyzing the scanning and imaging with image processing software installed in an HPLCM system; obtaining the glass microprobe which is suitable for performing whole cell recording by adopting a patch clamp technology; recording the ion channel current of the living neuron-like PC12 cell under physiological conditions by adopting the patch clamp technology; and computing corresponding inward current and current density by using the ion channel current of the PC12 cell. The invention has the advantages: high operability is achieved by combining the HPLCM technology with the patch clamp technology, and the neuron-like differentiation degree of the PC12 cell can be qualitatively, quantitatively and comprehensively analyzed and evaluated more accurately on the aspects of structures and functions.

Description

(1) Technical field [0001] The present invention belongs to the field of evaluating the degree of cell differentiation, especially a device and method for evaluating the degree of neuron-like differentiation of PC12 cells, especially refers to comprehensively evaluating the neuron-like differentiation of PC12 cells from the aspects of structure and function of living cells, and more Specifically, based on the constantly changing surface microtopography and electrophysiological characteristics of PC12 cells under the continuous induction and differentiation of nerve growth factor (NGF), probe-hopping mode scanning ion conductance microscopy (HPICM) and its combination The patch clamp technique, a method for assessing the degree of neuron-like differentiation of PC12 cells by identifying their corresponding structural and functional characteristic parameters. (2) Background technology [0002] PC12 cells are rat adrenal pheochromocytoma cells, and neuron cells are derived from...

Claims

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Application Information

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IPC IPC(8): G01Q60/44
CPCG01Q60/44
Inventor 张彦军刘晓
Owner CHINA NAT ACAD NANOTECH & ENG
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