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Electro-chemistry detecting method for fast identifying peripheral nerve bundle nature

A peripheral nerve and detection method technology, applied in biochemical equipment and methods, microbial determination/inspection, measuring devices, etc., can solve the problems of in-situ staining, clinical application limitations, long staining time, etc., and achieve simplified steps, fast identification, low-cost results

Inactive Publication Date: 2008-08-20
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the staining time of this method is long, the process is complicated, and in situ staining cannot be performed, so the clinical application is also limited.

Method used

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  • Electro-chemistry detecting method for fast identifying peripheral nerve bundle nature
  • Electro-chemistry detecting method for fast identifying peripheral nerve bundle nature
  • Electro-chemistry detecting method for fast identifying peripheral nerve bundle nature

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1a

[0031] Before the experiment, the gold plate electrode (4) was polished on silicon carbide fine sandpaper, and then polished into a "mirror surface" with suede with 0.3 μm and 0.05 μm alumina suspension, and finally cleaned by ethanol and secondary water ultrasonically. ,spare.

[0032]1. In the process of modifying the electrode, first fix the gold electrode in the spin coater with the surface facing up, and mix 0.1mL of 20% polydimethyldiallylamine aqueous solution with a mass ratio of 7:10:1 (weight-average molecular weight 100000, Aldrich company), polydimethylsiloxane (weight-average molecular weight 60000) and curing agent (Sylgard 184 Dow Coming company) are drip-coated on the electrode surface, start the spin coater, at 2000 rpm Rotate at a high speed for 15 minutes, and then stand in an oven at 80°C for one hour to solidify the polymer film on the electrode surface.

[0033] 2. Immerse the gold electrode modified polymer film into a beaker containing 0.5% chloroauric...

Embodiment 1b

[0039] Before the experiment, the gold plate electrode (4) was polished on silicon carbide fine sandpaper, and then polished into a "mirror surface" with suede with 0.3 μm and 0.05 μm alumina suspension, and finally cleaned by ethanol and secondary water ultrasonically. ,spare.

[0040] 1. In the process of modifying the electrode, first fix the gold electrode in the spin coater with the surface facing up, and mix 0.1 mL of 20% polydimethyldiallylamine ( Weight-average molecular weight 20000, Aldrich company) aqueous solution, polydimethylsiloxane (weight-average molecular weight 60000) and curing agent (Sylgard 184 Dow Corning company) are drip-coated on the electrode surface, start the spin coater, at 2500 rpm Rotate at a high speed for 15 minutes, and then stand in an oven at 80°C for one hour to solidify the polymer film on the electrode surface.

[0041] Other steps are the same as Example 1a, and the result obtained is exactly the same as Example 1a.

Embodiment 2

[0043] 1. The Beagle dogs were anesthetized and killed in stages, and the spinal canal was opened from the middle of the back immediately, and the T section running from the epidural to the entrance of the spinal nerve root canal was taken. 12 -S 2 For the anterior root and posterior root of the spinal nerve (the anterior root is the motor nerve, and the posterior root is the sensory nerve), fresh specimens of the two types of nerve roots of each dog were put into specimen bottles, and quickly stored in liquid nitrogen for future use. Naturally thaw at room temperature when taking it, and store it in a wet box after thawing to prevent the specimen from drying and dehydrating.

[0044] 2. Immerse a nerve specimen in 0.05M 1mL pH7.4 phosphate buffer solution and break it up in a tissue homogenizer. Then centrifuge at 10,000 rpm at 4°C for 15 min, take the supernatant and add ammonium sulfate to 25% saturation. Afterwards, centrifuge at 10000 rpm at 4°C for 15 min, remove the p...

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Abstract

The invention provides an electrochemical detection method for fast identifying peripheral nerve bundle characteristic, which is an electrochemical detection method for qualitatively detecting acetylcholinesterase in peripheral nerve bundle which is based on detecting oxidative signal on electrode surface produced by enzyme reaction products. The method has the following steps: first using a polymer gold nanoparticle complex to modify electrode, which is then incubated in choline oxidase solution and acetylcholinesterase solution orderly, through good adsorption force of protein on complex film, the two kinds of enzymes enrich on the electrode surface, through three-electrode system, detecting the ampere response of substrate acetylcholine based on two enzymes system, then the detection of acetylcholinesterase is indirectly realized (detecting concentration range is: 1.0-2.0unit / mL). The invention is a breakthrough in the field of identifying clinical differential peripheral nerve bundle characteristic for incubating in nervous homogenates instead of standard acetylcholinesterase solution, according to whether acetylcholine has response, it realizes fast identification of motor tract and sensory tract.

Description

technical field [0001] The present invention relates to a method for electrochemical identification of the properties of peripheral nerve bundles. Background technique [0002] In nerve repair after peripheral nerve injury (such as replantation of severed limbs), one of the difficulties is that the motor bundle and sensory bundle in the nerve trunk cannot be correctly distinguished during the operation. The existing suture methods such as epineurium and perineurium suture Legally, because the correct suture of the same nerve bundle at the two stumps cannot be realized, the recovery of the patient's neurological function is seriously affected. If the sensory bundles at the two stumps of the nerve can be realized. The correct suture of motor bundle to motor bundle will greatly improve the recovery rate of nerve function. The enzymatic histochemistry method currently used is based on the difference in the content of a neurotransmitter enzyme, acetylcholinesterase, in the motor...

Claims

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Application Information

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IPC IPC(8): G01N27/416G01N27/327C12Q1/46
Inventor 赵微徐静娟陈洪渊曹晓建
Owner NANJING UNIV
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