CN102805664A

CN102805664A - Method for displaying and acquiring innervation image in human body levator ani muscle

Info

Publication number: CN102805664A
Application number: CN2012103073170A
Authority: CN
Inventors: 王文艳; 卫兵; 张妤; 范懿隽; 周晓
Original assignee: 王文艳
Priority date: 2012-08-27
Filing date: 2012-08-27
Publication date: 2012-12-05

Abstract

The invention discloses a method for displaying and acquiring an innervation image in human body levator ani muscle. The method comprises the following steps of: dyeing nerves in a levator ani muscle specimen by using a sihler nerve dyeing method; manufacturing a hollow transparent silica gel model and placing a background light source in the silica gel model; placing the levator ani muscle specimen which is subjected to nerve dyeing on the outer surface of the silica gel template and acquiring a stereoscopic shape image of the innervation in the levator ani muscle by using a camera under irradiation of the background light source; placing the levator ani muscle specimen which is subjected to nerve dyeing under a dissecting microscope; tracing the running direction of nerves along the nerve trunk in the levator ani muscle under the dissecting microscope; observing a fine structure for connection between nerves in the specimen under the microscope; and recording by using an image acquisition system connected to the microscope. According to the method provided by the invention, the levator ani muscle is in a transparent state and the nerves in the levator ani muscle are violet blue, and further the aim of completely displaying the innervation state in the levator ani muscle is fulfilled under the mapping of the background light source.

Description

Show and gather the method for neural distributed image in the human anus elevator

Technical field

The present invention relates to neural staining technique, specifically a kind of method that shows and gather neural distributed image in the human anus elevator.

Background technology

Levator ani m. is the thin layer muscular tissue that is positioned at nervus vasculairs clump below, pelvis deep and ischiorectal fossa top; The levator ani m. pathological changes is to cause that pelvic floor dysfunction property disease (comprises urinary incontinence; Pelvic organ prolapse; Fecal incontinence, chronic pelvic pain) main cause of clinical disease such as, the nerve in the clear and definite levator ani m. are distributed with the pathogenesis that helps to find cause the neurogenic disease of levator ani m. pathological changes; Reduce the chance of the blind method paracentesis of Wicresoft, reduce because the related complication that the levator ani m. nerve injury causes to neural trunk damage in the levator ani m..

Because the position of levator ani m. is special, composition is complicated (The irregular shape muscle that levator ani m. is made up of puborectalis, musculus pubococcygeus and illiococcygeous; Part puborectalis and male urethra or female urethra and perivaginal sphincter merge), big to the research difficulty of levator ani m. and the outer traveling of dominating neural intramuscular thereof.At present, the research to levator ani m. in the anatomical research is less, and a lot of scholars attempt to adopt research methoies such as gross anatomy, iconography, SABC, structure animal model that in-depth explanation is carried out in source of the nerve in the levator ani m. and neural distribution mode.Cause that defectives such as skew take place in the neural blood vessel position of the region of anatomy owing to exist the little nervous ramification of damage, little nervous ramification and minute blood vessel and fascia to differentiate in unclear, the dissection process in the gross anatomy process, the result who therefore studies to the macroscopical anatomy of nerve in the levator ani m. often there are differences; Though iconography research can objectively respond the muscular tissue form, can't show the neural direction of travel in the muscular tissue; SABC and immunofluorescence technique only can show the details of the local tomography of BIAO and BEN, can't sketch the contours of the configuration and the inner nerve trend of BIAO and BEN of BIAO and BEN; Use the apes mammal the most close when making up animal model, also can't be equal to human organizational structure fully and distribute with neural with the human tissue structure.Above-mentioned research method all can not be under the situation of not destroying the BIAO and BEN integrity simultaneously, the whole three-dimensional distribution that shows nerve in the levator ani m..

Because levator ani m. and internal organs such as urethra, vagina, rectum and anal canal are in close relations; Clinical scholar's mistake of most department of obstetrics and gynecology, Urology Surgery and anorectal surgical is assigned levator ani m. as internal organs muscle; Rather than be the part that skeletal muscle is formed entirely; Be short of the Professional knowledge of the neural colouring method of anatomy aspect simultaneously owing to the clinical scholar of relevant speciality, and less in the classical basic anatomical research to the research of levator ani m. and dominating neural thereof.Though the neural colouring method of Sihler is the anatomy colouring method that a kind of relatively more classical research skeletal muscle intrinsic nerve distributes, this method also only is mainly used in neural Research on Distribution in the body surface skeletal muscle (comprising extremity and musculus pectoralis abdominis meat).So far do not have and adopt the report of the neural colouring method of sihler to the neural research that distributes in the levator ani m..

Summary of the invention

The object of the present invention is to provide a kind of method that shows and gather neural distributed image in the human anus elevator, this method can obtain distributed image neural in the levator ani m. exactly under the situation of not destroying the BIAO and BEN integrity.

For realizing above-mentioned purpose, the method for neural distributed image is in demonstration provided by the invention and the elevator of collection human anus: adopt the neural colouring method of sihler that the nerve in the human anus elevator BIAO and BEN is dyeed; Make the silica gel model of hollow printing opacity, at the inner background light source of placing of silica gel model; To accomplish neural painted levator ani m. BIAO and BEN and be placed on silica gel model outer surface, under the background light source irradiation, gather the neural solid shape image that distributes in the levator ani m. with photographing unit; To accomplish neural painted levator ani m. BIAO and BEN is placed under the anatomic microscope; Neural trunk is reviewed neural direction of travel in the levator ani m. under anatomic microscope; The fine structure that connects between nerve in the BIAO and BEN under the sight glass, and with the image capturing system record that is connected on the microscope.

Demonstration provided by the invention and the method for gathering neural distributed image in the human anus elevator can make levator ani m. be pellucidity and nerve in the levator ani m. is hyacinthine; And then under the mapping of background light source; Reach the purpose of neural distribution in the complete demonstration levator ani m., for the traveling regularity of distribution neural in the clear and definite human anus elevator provides effective research method.

The method of neural distributed image has following good effect in demonstration provided by the invention and the elevator of collection human anus:

1, can be from different viewed, understand the position of nervous ramification in the levator ani m. and the regularity of distribution of neural trunk and teleneuron accurately, for anatomy, physiology and clinical medical correlational study provide accurately the neural theoretical basis that distributes in the levator ani m..

2, the neural regularity of distribution in the clear and definite levator ani m. for the function of understanding levator ani m., carry out neural Electromyographic research in the levator ani m., the neural operation of Wicresoft has important directive significance at the bottom of the development innovation basin.

3, further understand maternal infuries, physiological and pathologicals such as amyotrophy change levator ani m. and the neural influence of intramuscular, and for reducing, delay and treat maternal infuries, physiological and pathologicals such as levator ani m. atrophy sexually revise at the bottom of the basin that causes disease such as organ prolapse provides research basic.

4, be to inquire into because the unusual pathogenesis of urinary tract, reproductive tract and function of intestinal canal that levator ani m. function and textural anomaly cause provides anatomical basis.

The specific embodiment

Below further specify the present invention through a specific embodiment.

At first, the nerve in the human anus elevator BIAO and BEN is dyeed may further comprise the steps:

1, collects the fresh cadaver BIAO and BEN.

2, formalin fixed: adopting concentration is that 10% formalin solution soaks fixedly 4～8 weeks of pelvis BIAO and BEN, changes formalin solution weekly once.

3, specimen sampling: remove pelvic cavity viscera, complete excision pelvic diaphragm muscle and soft tissue along the pelvis medial wall are separated and are cut the levator ani m. BIAO and BEN.

4, softening fading: it is softening fading for 2～3 weeks in 3% the KOH solution that BIAO and BEN is soaked into concentration, changes KOH solution weekly once, is transparent or semitransparent shape until BIAO and BEN.In addition, 3 concentration of use normal burette adding are 3% hydrogen peroxide in the KOH solution of the every 100ml that is adopted.

5, decalcification: BIAO and BEN is soaked into 2～4 weeks in the Sihler I solution, changes Sihler I solution weekly once, the BIAO and BEN muddiness that causes until decalcification becomes transparent or semitransparent once more.

6, dyeing: BIAO and BEN is soaked into 3～4 weeks in the Sihler II solution, changes Sihler II solution weekly once, dyed darkviolet until BIAO and BEN.

7, decolouring: BIAO and BEN was immersed in the Sihler I solution 2～4 hours, and per hour observation under anatomic microscope is lavender until muscle, and nerve fiber is bluish violet, and tiny teleneuron is clear and legible.

8, neutralization: it is 0.05% lithium carbonate solution 1～2 hour that BIAO and BEN is immersed concentration, changes lithium carbonate solution per half an hour one time, does not have obvious variable color until soak, and nerve fiber dyeing is deepened.

9, purification: BIAO and BEN was immersed in 40%, 60%, 80% and 100% glycerite each 24 hours successively, make painted muscle color thin out, nerve fiber is at printing opacity irradiation demonstration more clear down.

10 is transparent: will accomplish neural painted BIAO and BEN and soak preserve with 100% glycerol, and the transparency of BIAO and BEN is improved and strengthen.

The above-mentioned Sihler I solution that adopts is by the glacial acetic acid of 1 volume, and the glycerol of 1 volume and 6 volumes, 1% chloral hydrate aqueous solution are formed; The above-mentioned Sihler II solution that adopts is by 1 volume EhrlichShi hematoxylin, and the glycerol of 1 volume and 6 volumes, 1% chloral hydrate aqueous solution are formed.In addition, before carrying out above-mentioned each operating procedure, all need wash the soak on the BIAO and BEN with flowing water.

After accomplishing above-mentioned steps, can carry out the following work of gathering substantially and dissecting the image of microscopically:

(1), make hollow printing opacity silica gel model, the outer surface of model is a curved surface, like sphere or parabola, model is hemispherical or is similar to hemispherical ,At the inner background light source of placing of silica gel model; Background light source adopts electric filament lamp; To accomplish neural painted levator ani m. BIAO and BEN and be placed on the silica gel model surface, and under the background light source irradiation, gather the neural solid shape image that distributes in the levator ani m. with photographing unit, photographing unit adopts single anti-digital camera.

(2), will accomplish neural painted levator ani m. is placed under the anatomic microscope; Neural trunk is reviewed neural direction of travel in the levator ani m. under anatomic microscope; The fine structure that connects between nerve in the BIAO and BEN under the sight glass, and with the image capturing system record that is connected on the microscope.

After accomplishing above work, can understand the neural whole characteristics that distribute with traveling in the levator ani m., draw out the interior neural scattergram of levator ani m., and be applied to clinical and basic research such as dissection and physiology in.

Claims

1. show and gather the method for neural distributed image in the human anus elevator, it is characterized in that may further comprise the steps: adopt the neural colouring method of sihler that the nerve in the human anus elevator BIAO and BEN is dyeed; Make the silica gel model of hollow printing opacity, at the inner background light source of placing of silica gel model; To accomplish neural painted levator ani m. BIAO and BEN and be placed on silica gel model outer surface, under the background light source irradiation, gather the neural solid shape image that distributes in the levator ani m. with photographing unit; To accomplish neural painted levator ani m. BIAO and BEN is placed under the anatomic microscope; Neural trunk is reviewed neural direction of travel in the levator ani m. under anatomic microscope; The fine structure that connects between nerve in the BIAO and BEN under the sight glass, and with the image capturing system record that is connected on the microscope.

2. demonstration according to claim 1 and the method for gathering neural distributed image in the human anus elevator, it is characterized in that the nerve in the human anus elevator BIAO and BEN dyeed may further comprise the steps:

(1), to use concentration be 10% formalin fixed human pelvis BIAO and BEN, global solution cuts the levator ani m. BIAO and BEN along the pelvis medial wall;

(2), the levator ani m. BIAO and BEN is soaked into concentration is softening fading in 3% the KOH solution, is transparent or semitransparent shape until BIAO and BEN;

(3), the levator ani m. BIAO and BEN is soaked into decalcification in the Sihler I solution, the BIAO and BEN muddiness that causes until decalcification becomes transparent or semitransparent once more;

(4), the levator ani m. BIAO and BEN is soaked in the Sihler II solution dyes, dyed darkviolet until BIAO and BEN;

(5), the levator ani m. BIAO and BEN immersed in the Sihler I solution decolours, be bluish violet until nerve fiber, tiny teleneuron is clear and legible;

(6), the levator ani m. BIAO and BEN is immersed concentration is to accomplish N-process in 0.05% the lithium carbonate solution, does not have obvious variable color until soak, nerve fiber dyeing is deepened;

(7), the levator ani m. BIAO and BEN is immersed in 40%, 60%, 80% and 100% glycerite successively carry out purification;

(8), will accomplish neural painted BIAO and BEN and soak preservation with 100% glycerol.

3. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator is characterized in that: in said step (1), adopt formalin solution to soak fixedly 4～8 weeks of pelvis BIAO and BEN, change formalin solution weekly one time.

4. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator; It is characterized in that: in said step (2); 3 concentration of use normal burette adding are 3% hydrogen peroxide in the KOH solution of every 100ml; The levator ani m. BIAO and BEN is soaked into softening fading for 2～3 weeks in the KOH solution, changes KOH solution weekly once.

5. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator, it is characterized in that: in said step (3), the levator ani m. BIAO and BEN is soaked into 2～4 weeks in the Sihler I solution, changes Sihler I solution weekly once.

6. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator, it is characterized in that: in said step (4), the levator ani m. BIAO and BEN is soaked into 3～4 weeks in the Sihler II solution, changes Sihler II solution weekly once.

7. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator is characterized in that: in said step (5), the levator ani m. BIAO and BEN immersed in the Sihler I solution 2～4 hours, changed Sihler I solution per half an hour once.

8. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator is characterized in that: in said step (6), the levator ani m. BIAO and BEN immersed lithium carbonate solution 1～2 hour, and change lithium carbonate solution once per half an hour.

9. demonstration according to claim 2 and the method for gathering neural distributed image in the human anus elevator is characterized in that: the levator ani m. BIAO and BEN immersed in 40%, 60%, 80% and 100% glycerite respectively 24 hours successively in said step (7).