The Experimental Study Of Rapid Immunohistochemical Methods Identified Human Peripheral Nerve Fiber

Objective: It is well known that the peripheral nerves is composed of motor and sensory fibers, neural function is by these two kinds of nerve bundles of proportion of the decision. Nerve fibers and not exist alone. They each combination and nerve bundle membrane separates nerve fibers composition bundle of nerves. Neural stem function beam distribution and nature of the judge to guide the choice of surgical methods of clinical. The current distinction between nerve bundles, because there are a lot of their own deficiencies and shortcomings in the clinical practice it is difficult. Since 1970, people found many neurons specific protein in immunohistochemical method under the help of the people, the development process of neurons have further understanding. Immunohistochemical techniques (immunohistochemistry) is the basic principle, application of immunology antigen-antibody reaction antigen and antibody specificity, namely, through the theory of chemical reaction to mark the antibody colour-display reagent (fluorescent element, enzyme, metal ions, isotopes) color to determine the organization intracellular antigens (peptide and protein), carries on the orientation, qualitative and quantitative research and immune cells called chemical technology (immunocytochemistry). Although immunohistochemical technology has advantages, but the key is how to determine the sensory nerve and nerve characteristic marker, through the literature review, this experiment target in the membrane protein V, hoped through al immunohistochemical techniques to reveal membrane proteins in peripheral nerve V al the characteristics, so as to achieve the objective identification nerve bundle nature. Membrane protein (al Annexin V) is a kind of phospholipids on calcium ions, molecular binding proteins for 35KD, it has many functions such as anticoagulation, anti-inflammatory, calcium ion channels, in peripheral nerve in Annexin V mainly distributed in sensory nerve of axons, not found in the motor. This experiment by means of fast yow-chyi method to confirm the specificity of membrane protein V al, and using the characteristics of this application from the fault of peripheral nerve, hope to achieve rapid identify clinical guidelines of peripheral nerve function, thus establishing the scientific experiment and clinical application of Bridges.Methods: Experiment 1.Choose normal healthy male SD rats, weight only 250±10g, Cut out rat spinal nerve root 5mm, before after spinal nerve root 5mm, exposure and siatic nerve and muscle and skin, 5mm, lumbav immediately above freezing crosscut slice organization, thick slices 5um ,Starting SABC immunohistochemical experiment, Nerve biopsy via the antigen-antibody reaction - DAB color - hematoxylin-eosin stain, pure complex, transparent, wafer processing, microscope color, By using the substitution test group, an antioxidant, the substitute PBS experimental procedure. Experiment 2 Taking fresh abandoned landmine forearm ulnar nerve trunk, ulnar wrist back teams (skin) and the median nerve big teams each 5mm. Yu muscle,the organization immediately frozen section, thick slices 5um, Starting S-P immunohistochemical experiment, nerve biopsy via the antigen-antibody reaction - DAB color - hematoxylin-eosin stain, pure complex, transparent, wafer processing, microscope color, By using the substitution test group, an antioxidant, the substitute PBS experimental procedure.Results: Experiment 1 SD rat spinal nerve root, with a negative response, before the heel for positive reaction to dye, muscle teams of nerve fibers, skin with hyperchromatic teams, including positive nerve fibers sciatic nerve fibers in both positive and negative has the nerve fiber dyed, at high magnification observation, the nerve fiber immune dyeing of axons, myelin not colouring. The glial tissue is a negative response.Experiment 2 The median nerve, with thenar of nerve fibers, its feet nerve with hyperchromatic wrist back a nerve fiber, the positive one nerve fibers in both positive and negative has the nerve fiber dyed, ulnar nerve trunk, The glial tissue is a negative response . Conclusion: We know that in the distal nerve dry, dry branches in the nerve in positioning, which has specific functions in the body, and girded proximal, each bundle of nerves, no single function of mixed bundle. In the mix of fiber bundle of nerves, the proportion of differences. If a large proportion of fiber in axonal regeneration, into the distal nerve endometrial tube process, will have more opportunities of endometriosis neural tube into the related peripheral nerve injury, thus breaking in situ, nerve or shift, the nerve repair when transplanted neural stem in understanding performer fine structure, especially within the neural function of the nerve stem of positioning knowledge, so that it was possible to precise selective nerve bundle branch, reducing regeneration fibers showed the vagus caused by void regeneration, improve the recovery rate of postoperative limb function. Therefore the clinical needs a rapid, accurate identification of sensory nerve bundle around and nerve bundle method, this experiment through membrane proteins in peripheral nerve V al organizational characteristics, broke the immune method identified no exact molecular properties bundle of nerves, and the bottleneck of the method is applied to human peripheral nerve, validated in human peripheral nerve identification method of peripheral nerve bundle for feasibility, provides a new means to identify function, this kind of method applies early clinical built bridge.