CN114052753A - Preparation method of tomographic specimen based on lung of viral pneumonia patient - Google Patents
Preparation method of tomographic specimen based on lung of viral pneumonia patient Download PDFInfo
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Abstract
The invention provides a preparation method of a tomographic specimen based on the lung of a viral pneumonia patient, which is characterized in that a corpse is placed in a splash-proof autopsy isolating device for processing in a biological safe autopsy room, and the preparation method comprises the following steps: 1) dissecting the corpse, taking out the lung, soaking the lung in a fixing solution, and taking out the lung to obtain a lung specimen; 2) positioning a horizontal cross section datum line on the surface of the lung specimen by adopting a line snapping method, then scribing a saw line, and freezing the lung specimen; 3) sawing the frozen lung specimen along a sawing line, and verifying by CT scanning to obtain a fault specimen; 4) and (3) carrying out pathological material taking, cleaning and glue brushing on the tomographic specimen, then carrying out imaging treatment, and sealing and storing. The invention provides a preparation method of a tomographic specimen based on the lung of a viral pneumonia patient, which reserves precious clinical pathological data for the subsequent study of viral pneumonia and avoids unnecessary infection of an operator in the manufacturing process.
Description
Technical Field
The invention belongs to the field of medical biotechnology, relates to a preparation method of a tomographic specimen of a lung of a patient with viral pneumonia, and particularly relates to a preparation method of a clinical diagnosis, scientific research and teaching specimen of a tomographic specimen of a lung of a patient with viral pneumonia (especially pneumonia with high infectivity, such as novel coronavirus pneumonia, SARS and the like).
Background
Viral pneumonia, especially viral pneumonia with high infectivity, has become an infectious disease that seriously threatens the health and life of the whole human. In the future, viral pneumonia is likely to become a public health disease which troubles human society for a long time. Therefore, it is necessary to store as many cases of viral pneumonia as possible and to fully study their imaging and pathological conditions in order to lay a solid foundation for better understanding of this infectious disease.
Advances in medicine are not isolated from pathological anatomy. The pathology can reveal the nature of the disease, including the etiology, pathogenesis, and changes and laws in organs, tissues, cells, and even molecules of the body during the course of the disease, which are the basis of the origin and development of modern medicine. The pathological research result of the viral pneumonia obtained by dissection has great help for understanding the pathological change and essence of the viral pneumonia, disclosing the occurrence mechanism and development rule and perfecting the clinical diagnosis and treatment scheme, and can fundamentally search the pathogenic and lethal reasons of the viral pneumonia and provide basis for clinically curing critically ill patients.
Pathology is the basis of imaging, which is the manifestation of the pathology. The major target organ involved in viral pneumonia is the respiratory system, mainly the lung parenchyma. However, due to the lack of safety-compromised necropsy sample preparation methods, there is still a lack of standardized procedures available to follow for viral pulmonary specimens for pulmonary inflammation. More importantly, the current research and diagnosis of viral pneumonia are mostly in a state of splitting the pathological morphology from the clinical image or only slightly corresponding to the pathological morphology, and a research scheme for deeply fusing the two is lacked. Therefore, for the lung of patients with viral pneumonia, it is urgently needed to perform contrast research of highly fusing clinical images and anatomical pathologies, establish a viral pneumonia database, and manufacture clinical scientific research and teaching specimens integrating clinical anatomy, pathology and images.
Tomosynthesis is a science that utilizes the dissection saw method to study the morphological structure of the human body. On the basis of maintaining the structure of the machine body in situ, the method can accurately present the form change of the section and the position adjacent relation of each structure of the machine body; three-dimensional reconstruction and quantitative analysis of the structure by tracking the continuous fault or by means of a computer; it closely combines image diagnostics and interventional radiology, and is the product of anatomical and medical imaging. The lung of a patient with viral pneumonia is subjected to tomosynthesis diagnosis and pathological diagnosis, and morphological basis can be provided for teaching, scientific research and image diagnosis of the diseases.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a preparation method of a viral pneumonia patient lung-based tomographic specimen, the viral pneumonia lung cross section tomographic specimen prepared by the method can fully meet the requirements of clinical diagnosis, teaching and scientific research of images and pathology, and each tomographic specimen realizes mutual correspondence of image data and pathological data; moreover, because the manufactured tomographic specimen is clean and tidy and is fully fixed by neutral formalin, the tomographic specimen can be stored for a long time, is visual and vivid when being used for teaching, scientific research and clinical image diagnosis, and can retain precious clinical pathological data for human beings to study viral pneumonia in the future.
In order to achieve the above objects and other related objects, a first aspect of the present invention provides a method for preparing a tomographic specimen based on lungs of a patient with viral pneumonia, wherein a corpse is disposed in a splash-proof autopsy isolation device for processing in a biosafety autopsy room, comprising the steps of:
1) dissecting the corpse, taking out the lung, soaking the lung in a fixing solution, and taking out the lung to obtain a lung specimen;
2) performing horizontal cross section datum line positioning on the surface of the lung specimen obtained in the step 1) by adopting a line snapping method, then scribing a saw line, and freezing the lung specimen;
3) sawing the lung specimen frozen in the step 2) along a sawing line, and verifying by CT scanning to obtain a fault specimen;
4) carrying out pathological material taking, cleaning and glue brushing on the fault specimen obtained in the step 3), then carrying out imaging treatment, and sealing and storing.
Preferably, the corpse is placed in a splash-proof autopsy isolation device for treatment, and the method comprises the following steps:
a) the splash-proof autopsy isolating device comprises a pollution source chamber and a semi-clean chamber which are isolated by an isolating surface, a curtain door on one side surface of the pollution source chamber adjacent to the isolating surface is opened, a corpse is placed in the pollution source chamber, and after the curtain door is closed, the corpse is dissected through at least one first operation port arranged on the other side surface adjacent to the isolating surface and/or at least one second operation port arranged on the side surface opposite to the isolating surface, and the lung is taken out;
b) a second isolation lifting curtain is arranged on the side surface, opposite to the isolation surface, of the semi-cleaning chamber, and the lung is transferred to the semi-cleaning chamber from the pollution source chamber through the first isolation lifting curtain arranged on the isolation surface through at least one third operation port arranged on the second isolation lifting curtain;
c) placing the lung in a specimen storage container arranged in the semi-cleaning chamber, soaking the lung in a fixing solution to obtain a lung specimen, scribing and sawing the lung specimen, opening a second isolation lifting curtain, taking out the lung specimen, and freezing;
d) placing the frozen lung specimen into a semi-clean chamber through a second isolation lifting curtain, performing saw cutting through a third operation port to obtain a fault specimen, performing pathological material taking, cleaning and glue brushing on the fault specimen, sealing, and taking out the fault specimen through the second isolation lifting curtain for imaging treatment;
e) and putting the imaged fault sample into a semi-clean chamber through a second isolation lifting curtain to manufacture a specimen box, and taking out the specimen box.
Preferably, the splash necropsy preventing isolation device comprises a pollution source chamber and a semi-clean chamber which are adjacent, the pollution source chamber and the semi-clean chamber are isolated by an isolation surface, and a first isolation lifting curtain is arranged on the isolation surface; the side surface of one side of the pollution source chamber adjacent to the isolation surface is a curtain door, the side surface of the other side of the pollution source chamber adjacent to the isolation surface is provided with at least one first operation port, and the side surface opposite to the isolation surface is provided with at least one second operation port; and a second isolation lifting curtain is arranged on the side surface of the semi-cleaning chamber opposite to the isolation surface, and at least one third operation port is arranged on the second isolation lifting curtain.
More preferably, a top plate is arranged at the top of the pollution source chamber, and a plurality of support rods are arranged below the top plate.
Further preferably, 4 support rods are arranged below the top plate.
More preferably, the first isolation lifting curtain, the second isolation lifting curtain and the curtain door are of an upper opening type openable and closable structure.
More preferably, the number of the first operation opening and/or the second operation opening is 2.
More preferably, the number of the third operation ports is 2.
More preferably, the first operation opening, the second operation opening and the third operation opening are provided with operation gloves with sealed interfaces.
More preferably, a specimen storage container is arranged in the semi-cleaning chamber.
Preferably, in step 1), the cadaver is a cadaver of a patient with viral pneumonia. The viral pneumonia includes, but is not limited to, novel coronavirus pneumonia or SARS.
Preferably, in step 1), before the corpse is placed in the splash-proof autopsy isolation device, the corpse is shaved to remove hairs and oil stains on the skin of the corpse, and after the corpse is sterilized by a disinfectant, the corpse is fixed by a bandage.
More preferably, the disinfectant is 70-80% ethanol water solution by volume percentage. Most preferably, the disinfecting liquid is 75% ethanol water solution by volume percentage.
More preferably, the cadaveric typing is performed according to an anatomical pose.
Preferably, in the step 1), the lung is dissected and taken out, the trachea of the cadaver is cut off, the chest cavity is incised by adopting a T-shaped incision method, the sternum and costal cartilage are removed, the chest cavity is exposed, the artery, the trachea, the lung root and the lung ligament are cut off, and the lung is taken out.
Preferably, in the step 1), the lung is soaked in the fixative by firstly infusing the fixative into the lung through the opening of the severed end of the trachea, and then soaking the lung filled with the fixative in the fixative.
More preferably, the fixative solution is a 5-15% neutral formalin solution. Most preferably, the fixative solution is a 10% neutral formalin solution.
More preferably, the lungs are soaked in the fixative for 46-50 h.
Preferably, in the step 1), the fixative is poured out from the lung specimen, the lung specimen is washed with clean water, the moisture on the surface of the lung specimen is absorbed, and then the lung specimen is kept stand and dried.
More preferably, the moisture on the surface of the lung specimen is absorbed by using absorbent filter paper.
More preferably, the standing and air-drying is to place the lung specimen with the surface moisture absorbed on the water-absorbing filter paper, and to stand the lung specimen in a shade place for 4 to 6 hours to air-dry the lung specimen.
Preferably, in step 2), the horizontal cross-section reference line is located with the longitudinal axis of the trachea as a coronal line and perpendicular thereto as a transverse line.
Preferably, in step 2), the titanium dioxide dye is titanium dioxide.
Preferably, in step 2), the scribing depth of the saw cutting line is 1-2 cm.
Preferably, in step 2), the lung specimen is sealed before freezing.
Preferably, in step 2), the freezing is performed by freezing the lung specimen at ≦ -30 ℃ for 4-6 days.
Preferably, in step 3), before the sawing, the lung specimen needs to be fixed, and the lung specimen should be fixed on a band sawing machine.
Preferably, in step 3), the sawing is performed by using a band sawing machine.
More preferably, the band saw machine has a saw wear of less than 2 mm.
More preferably, on the band sawing machine, the distance between the fixed baffle and the saw blade is equal to the thickness of the lung specimen to be sawed plus half of the saw consumption.
Preferably, in step 3), the thickness of the tomographic specimen coincides with the scanning thickness of CT.
More preferably, the thickness of the tomographic specimen and the scanning thickness of the CT are both 0.7-0.9 mm.
Preferably, in step 4), the pathological material taking comprises the following steps:
A) observing pathological changes of the fault specimen at room temperature, and taking lesion tissues for examination of a morphologically changed lesion area;
B) sequentially carrying out gradient alcohol dehydration, xylene transparence, paraffin wax immersion, paraffin wax embedding, pathological tissue section, slide sticking, tissue section xylene dewaxing, HE staining, gradient alcohol dehydration, xylene transparence and gum sealing treatment on the detected lesion tissues to obtain HE stained sections.
More preferably, in step a), said pathological change is selected from one or more of colour, texture, solid change, mucoid change.
Preferably, in step 4), the cleaning is rinsing with water.
Preferably, in the step 4), after cleaning, the moisture on the surface of the fault specimen is absorbed, and then the fault specimen is kept stand and dried.
More preferably, the water on the surface of the fault specimen is absorbed by using water absorption filter paper.
More preferably, the standing and air-drying is to place the fault specimen with the surface moisture absorbed dry on water-absorbing filter paper, and stand the fault specimen in a shade place for 4-6 hours to air-dry.
Preferably, in the step 4), the glue brushing is to brush the surface of the fault specimen with a gelatin solution, and then to stand and dry the surface.
More preferably, the gelatin solution is obtained by hydrolyzing collagen.
More preferably, the concentration of the gelatin solution is 15-25%.
More preferably, the brushing times of the gelatin solution are 1-3 times.
More preferably, the standing and airing are standing and airing in a shady and cool place.
Preferably, in step 4), the imaging process is to scan the two cross sections of the cleaned and brushed tomographic specimen by using a scanner, and then store the two cross sections in a hard disk of an electronic computer.
Preferably, in the step 4), the cleaned and brushed tomographic specimen is placed in a specimen box filled with preservation solution for permanent preservation.
More preferably, the preservation solution is a 4-6% neutral formalin solution.
More preferably, the cartridge is a polymethylmethacrylate plate cartridge.
In a second aspect, the present invention provides the use of the above method for the preparation of a tomographic specimen of the lungs of a patient with viral pneumonia.
As described above, the method for preparing a tomographic specimen based on the lung of a viral pneumonia patient according to the present invention has the following advantageous effects:
(1) the virus pneumonia lung cross section sectional specimen manufactured by the method can fully meet the requirements of clinical diagnosis, teaching and scientific research of two subjects of image and pathology, and each sectional specimen realizes the mutual correspondence of image data and pathological data.
(2) The virus pneumonia lung cross section sectional specimen prepared by the method of the invention is clean and tidy, can be preserved for a long time by fully fixing the specimen by neutral formalin, and reserves precious clinical pathological data for subsequent study of virus pneumonia.
(3) The preparation process of the virus pneumonia lung cross section specimen prepared by the method of the invention fully considers the infectivity of virus pneumonia, has obvious protection effect and avoids unnecessary infection of operators in the preparation process.
Drawings
Fig. 1 is a schematic view showing the overall structure of a splash-proof autopsy isolation device according to the present invention.
FIG. 2 shows an anatomical structure of a tomographic specimen of lung of a patient with viral pneumonia according to the present invention.
FIG. 3 is a view showing an HE stained section of a tomographic specimen based on the lung of a patient with interstitial pneumonia prepared in the present invention.
FIG. 4 shows an HE stained section of a lung sectional specimen of a critically ill patient with viral pneumonia prepared by the present invention.
Reference numerals
1 contamination Source Chamber
2 semi-clean chamber
3 first isolating lifting curtain
4-curtain door
5 first operation port
6 second operation port
7 second isolation lifting curtain
8 third operation port
9 operating glove
10 Top plate
11 support rod
12 specimen storage container
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and not to limit the scope of the invention.
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
It should be understood that the processing equipment or devices not specifically mentioned in the following examples are conventional in the art; all pressure values and ranges refer to relative pressures.
Furthermore, it is to be understood that one or more method steps mentioned in the present invention does not exclude that other method steps may also be present before or after the combined steps or that other method steps may also be inserted between these explicitly mentioned steps, unless otherwise indicated; it is also to be understood that a combined connection between one or more devices/apparatuses as referred to in the present invention does not exclude that further devices/apparatuses may be present before or after said combined device/apparatus or that further devices/apparatuses may be interposed between these two explicitly referred to devices/apparatuses, unless otherwise indicated. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.
The invention provides a preparation method of a tomographic specimen based on the lung of a viral pneumonia patient, which is characterized in that a corpse is placed in a splash-proof autopsy isolating device for processing in a biological safe autopsy room, and the preparation method comprises the following steps:
1) in a biological safe autopsy room, placing the autopsy in a splash-proof autopsy isolation device for treatment, dissecting and taking out the lung, soaking the lung in a fixing solution, and taking out the lung to obtain a lung specimen;
2) performing horizontal cross section datum line positioning on the surface of the lung specimen obtained in the step 1) by adopting a line snapping method, then scribing a saw line, and freezing the lung specimen;
3) sawing the lung specimen frozen in the step 2) along a sawing line, and verifying by CT scanning to obtain a fault specimen;
4) carrying out pathological material taking, cleaning and glue brushing on the fault specimen obtained in the step 3), then carrying out imaging treatment, and sealing and storing.
The biological safety autopsy room is a conventional autopsy room which meets the requirements of biological safety autopsy operation. The biosafety autopsy room can carry out negative pressure filtration, is provided with a special operation table, and also has shower or sanitary facilities, air-conditioning or cold and warm air equipment and a special sewage treatment system.
The invention provides a preparation method of a tomographic specimen based on the lung of a viral pneumonia patient, wherein the corpse is placed in a splash-proof autopsy isolating device for treatment, and the preparation method comprises the following steps:
a) the splash-proof autopsy isolating device comprises a pollution source chamber and a semi-clean chamber which are isolated by an isolating surface, a curtain door on one side surface of the pollution source chamber adjacent to the isolating surface is opened, a corpse is placed in the pollution source chamber, and after the curtain door is closed, the corpse is dissected through at least one first operation port arranged on the other side surface adjacent to the isolating surface and/or at least one second operation port arranged on the side surface opposite to the isolating surface, and the lung is taken out;
b) a second isolation lifting curtain is arranged on the side surface, opposite to the isolation surface, of the semi-cleaning chamber, and the lung is transferred to the semi-cleaning chamber from the pollution source chamber through the first isolation lifting curtain arranged on the isolation surface through at least one third operation port arranged on the second isolation lifting curtain;
c) placing the lung in a specimen storage container arranged in the semi-cleaning chamber, soaking the lung in a fixing solution to obtain a lung specimen, scribing and sawing the lung specimen, opening a second isolation lifting curtain, taking out the lung specimen, and freezing;
d) placing the frozen lung specimen into a semi-clean chamber through a second isolation lifting curtain, performing saw cutting through a third operation port to obtain a fault specimen, performing pathological material taking, cleaning and glue brushing on the fault specimen, sealing, and taking out the fault specimen through the second isolation lifting curtain for imaging treatment;
e) and putting the imaged fault sample into a semi-clean chamber through a second isolation lifting curtain to manufacture a specimen box, and taking out the specimen box.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, as shown in fig. 1, the splash-proof autopsy isolation device comprises a pollution source chamber and a semi-clean chamber which are adjacent to each other, wherein the pollution source chamber and the semi-clean chamber are isolated by an isolation surface, and a first isolation lifting curtain is arranged on the isolation surface; the side surface of one side of the pollution source chamber adjacent to the isolation surface is a curtain door, the side surface of the other side of the pollution source chamber adjacent to the isolation surface is provided with at least one first operation port, and the side surface opposite to the isolation surface is provided with at least one second operation port; and a second isolation lifting curtain is arranged on the side surface of the semi-cleaning chamber opposite to the isolation surface, and at least one third operation port is arranged on the second isolation lifting curtain.
The pollution source chamber and the semi-cleaning chamber are made of transparent plastic films. Because the pathological autopsy operation time is long, the exposed area of the visceral tissue section is large, a transparent plastic film which has good light transmittance and can resist repeated folding can be used, and the pathological autopsy scene can be suitable.
In a preferred embodiment, as shown in fig. 1, a top plate is disposed on the top of the contamination source chamber, and a plurality of support rods are disposed below the top plate. The bracing piece is used for supporting the pollution source cavity to drive simultaneously and support half clean cavity, form sufficient space and be convenient for the corpse to deposit in the pollution source cavity, and in the interior processing preparation sample of half clean cavity.
Specifically, 4 support rods are arranged below the top plate. The pollution source chamber can be stably supported, and enough space is kept.
The top plate and the support rod are made of transparent hard plastic. Has certain supporting capacity.
In a preferred embodiment, as shown in fig. 1, the first isolation opening curtain, the second isolation opening curtain and the curtain door are of an upper opening type openable and closable structure. The first isolation lifting curtain, the second isolation lifting curtain and the curtain door are opened from bottom to top.
The first isolation lifting curtain can prevent aerosol and body fluid generated during autopsy from splashing into the semi-cleaning chamber. Meanwhile, the human body specimen obtained after autopsy can be transferred to the semi-clean chamber for subsequent treatment by opening the first isolation lifting curtain.
The second isolation lifting curtain can block the possible situation that the fixing liquid splashes out of the isolation device when the fixing liquid preservation operation is carried out on the human body specimen. Meanwhile, the second isolation lifting curtain is opened, so that the human body specimen subjected to stationary liquid storage can be transferred to other places.
The openable curtain door can be opened and is used for placing corpses.
In a preferred embodiment, as shown in fig. 1, the first operation port 5 and/or the second operation port 6 are 2. The operator can conveniently carry out two-hand operation on the corpse of the pollution source chamber.
In a preferred embodiment, as shown in fig. 1, the number of the third operation ports is 2. The operator of being convenient for can carry out bimanualness to the human sample of half clean chamber.
In a preferred embodiment, as shown in FIG. 1, a specimen storage container is provided within the semi-clean chamber. The specimen storage container is a sealed tank filled with stationary liquid. Can prevent the cut human body specimen from becoming a pollution source in the moving process.
In a preferred embodiment, as shown in fig. 1, the first operation port, the second operation port and the third operation port are provided with operation gloves with sealing interfaces. The operating gloves can be convenient for the operator to carry out the gouging to the human sample of pollution source cavity and/or semi-clean cavity when, avoid the pollution source contact operator in pollution source cavity and/or the semi-clean cavity. Meanwhile, when the operation of the planer is not carried out, the first operation port, the second operation port and the third operation port can be closed.
In a preferred embodiment, as shown in FIG. 1, a specimen storage container is provided within the semi-clean chamber. The specimen storage container is a sealed tank filled with stationary liquid and can be used for placing the lung. Can prevent the cut human body specimen from becoming a pollution source in the moving process.
In the invention, the corpse is the corpse of the patient with viral pneumonia. Such viral pneumonia includes, but is not limited to, novel coronavirus pneumonia or SARS.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, before the corpse is placed in the splash-proof autopsy isolation device, the hair of the corpse is shaved off, dirt and oil stains on the skin of the corpse are removed, the corpse is disinfected by a disinfectant, and then the corpse is shaped by a bandage.
In a preferred embodiment, the disinfecting liquid is 70-80% ethanol water solution by volume percentage, preferably 75% ethanol water solution by volume percentage, so as to have the best sterilizing effect.
In a preferred embodiment, the cadaver shaping is performed according to an anatomical pose.
In the step 1), the lung is dissected and taken out, namely, the trachea of a dead body is cut off, the thoracic cavity is cut by adopting a T-shaped incision method, the sternum and costal cartilage are removed, the thoracic cavity is exposed, and then the artery, the trachea, the lung root and the lung ligament are cut off, so that the lung is taken out.
In a specific embodiment, the sternum and costal cartilage are removed by cutting open the thoracic cavity, removing the extrathoracic tissue, exposing the ribs, cutting off the costal cartilage from the second rib with a chondrotome, cutting off the sternoclavicular joint with a scalpel, cutting off the first ribs with rib scissors, lifting the costal arch, adhering the costal arch to the back of the sternum and costal cartilage, and separating the diaphragm muscle and mediastinum.
Wherein, when cutting off costal cartilage at two sides from the second rib, the tangential distance is 0.5-1cm from the boundary of costal cartilage and rib.
In one specific embodiment, the artery and trachea are cut off, namely the innominate artery and the left subclavian artery, and the trachea is cut off at the position 3-4cm above the trachea bifurcation by using a scalpel on the section of the rib cage cartilage.
In one embodiment, the lung root and ligaments are severed by separating the lung from the mediastinum, avoiding the vascular nerves around the lung root, and severing the lung root and ligaments vertically. After removal of the lung, attention was paid to the location and integrity of the bronchi, pulmonary arteries, pulmonary veins, bronchial arteries and bronchopulmonary lymph nodes at the hilum of the lung. Particularly, there are phrenic nerve and pericardial phrenic vessel in front of the left lung root, and vagus nerve behind; the accessory nerve and pericardial diaphragmatic vessel are in front of the right lung root, the vagus nerve is in the back, and the strange venous arch is above the right lung root and should be avoided.
In the step 1), the lung is soaked in the fixative, the fixative is firstly infused into the lung through an opening at the broken end of a trachea, and then the lung filled with the fixative is soaked in the fixative.
In a preferred embodiment, the fixative solution is a 5-15% neutral formalin solution, preferably a 10% neutral formalin solution.
In a preferred embodiment, the lungs are soaked in the fixative for 46-50 hours, preferably 48 hours.
When the perfusion fixative is put into the lung, the trachea is cut off at a position about 5cm away from the bifurcation of the trachea, a plastic tube of a disposable infusion apparatus wound with a gauze strip is inserted into the broken end of the trachea, the outer end of the plastic tube is aligned with the fracture of the trachea, gauze is lined between the plastic tube and the inner wall of the trachea, and the plastic tube and the trachea are fastened by a suture thread around the trachea. Sucking the fixative with an injector, injecting the fixative slowly from the center of the plastic tube, injecting the fixative in batches, if gas exists in the plastic tube during the injection process, slightly lifting the lung, simultaneously pulling out the injector, slowly putting down the lung to discharge the gas, and injecting the injector when the fixative is about to flow out, wherein the injection amount is not enough. The pressure is not too high during injection, so that the tube wall is prevented from being broken. If leakage is found, the hemostatic forceps can be used for clamping or the hemostatic forceps can be used for tying.
When the lung filled with the fixative is soaked in the fixative, the upper end (namely the trachea broken end) of the lung is fixed above the plastic barrel, and the normal anatomical position is kept.
In the preparation method of the fault specimen based on the lung of the viral pneumonia patient, in the step 1), the lung specimen is poured out of the stationary liquid, is washed by clean water, absorbs water on the surface of the lung specimen, and is then kept stand and dried.
In a preferred embodiment, the moisture on the surface of the lung specimen is absorbed by using absorbent filter paper.
In a preferred embodiment, the standing and air drying is to place the lung specimen with the surface moisture absorbed dry on a water-absorbing filter paper, and stand the lung specimen in a shade and dark place for 4 to 6 hours to air dry. During the period, the water-absorbing filter paper is timely replaced, the condition of the lung specimen in air drying is carefully observed, and the lung tissue is required to be kept to have certain humidity and cannot be dried too much.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, in the step 2), the horizontal cross section datum line is positioned by taking the longitudinal axis of the trachea as a coronary line, and the horizontal cross section datum line is marked as a cross section line perpendicular to the longitudinal axis of the trachea. The consistency of the fault specimen marking line and the lung longitudinal axis surface cross section positioning line is ensured.
In the method for preparing the section specimen based on the lung of the viral pneumonia patient, in the step 2), the line snapping method is a method which is conventionally used in the field. Specifically, a line stained with titanium white dye is used, one end of each of two persons is taken and then flicked on the surface of lung tissue, and actually, the line is used for replacing a pen, the titanium white dye is used for replacing ink, and the horizontal cross section datum line is positioned on the surface of the specimen.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, in the step 2), the titanium dioxide dye is titanium dioxide.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, in the step 2), the scribing depth of the saw line is 1-2 cm.
In the preparation method of the lung fault specimen based on the viral pneumonia patient, in the step 2), the lung specimen is sealed before freezing. Preventing infection.
In the preparation method of the lung fault specimen based on the viral pneumonia patient, in the step 2), the lung specimen is frozen at minus 30 ℃ or less for 4 to 6 days, preferably at minus 30 ℃ for 5 days.
In the preparation method of the lung fault specimen based on the viral pneumonia patient, in the step 3), the lung specimen needs to be fixed before sawing. In particular, the lung specimen should be mounted on a band saw machine. Specifically, the lung specimen is fixed on a fixed baffle of a band saw machine.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, in the step 3), the saw cutting is performed by a band sawing machine. The band saw machine is a joinery band saw machine which is used conventionally. Specifically, the band saw machine is a model MJ345E joinery band saw machine.
In a preferred embodiment, the band saw machine has a saw consumption of less than 2 mm. The band saw machine adjusts the tension of the saw blade according to different positions of the lung specimen to be sawed.
In a preferred embodiment, on the band sawing machine, the distance between the fixed baffle and the saw blade is equal to the thickness of the lung specimen to be sawed plus half of the saw consumption.
In the preparation method of the tomographic specimen based on the lung of the viral pneumonia patient, in the step 3), the thickness of the tomographic specimen is consistent with the scanning thickness of CT.
In a preferred embodiment, the thickness of the tomographic specimen and the scanning thickness of the CT are both 0.7-0.9mm, preferably 0.8 mm.
In the step 3), the length and the width of the fault specimen are carefully measured and are recorded one by one, the fault specimen is placed in a plastic bag, the serial number of the plastic bag is marked by a waterproof and oilproof pen, and the plastic bag is sequentially placed in a medical freezing freezer at the temperature of-30 ℃ for temporary storage. The specimen can not be placed in a high-temperature environment for too long time to prevent the specimen from being damaged.
In the step 4), the pathological material drawing comprises the following steps:
A) observing pathological changes of the fault specimen at room temperature, and performing lesion tissue sampling and detection on a pathological change area with morphological changes;
B) sequentially carrying out gradient alcohol dehydration, xylene transparence, paraffin wax immersion, paraffin embedding of the detected tissue, pathological tissue section, slide sticking, xylene dewaxing of the tissue section, HE staining, gradient alcohol dehydration, xylene transparence and gum sealing treatment on the detected lesion tissue to obtain an HE stained section.
In a preferred embodiment, in step a), said pathological changes include, but are not limited to, color, texture, solid changes, mucoid changes, and the like.
In step B), the above steps of gradient alcohol dehydration, xylene transparence, paraffin wax immersion, tissue paraffin embedding, pathological tissue section, slide sticking, tissue section xylene dewaxing, HE dyeing, gradient alcohol dehydration, xylene transparence, gum sealing treatment and the like are all conventional treatment means for lesion tissues, specifically refer to chapter iv "section making and dyeing technology" in "modern experimental pathological technology" (military medical science publishing, 8 months, 2012, version 1, main edition: penreous cloud and plum populus, vice-master edition: high soldiers and Liu super, main examination: wangden).
In step B), the HE stained sections are used for observing histopathological changes under a microscope.
In the preparation method of the section specimen based on the lung of the viral pneumonia patient, in the step 4), the cleaning is water washing.
In the step 4), after cleaning, the moisture on the surface of the tomographic specimen is sucked, and then the tomographic specimen is kept stand and dried.
In a preferred embodiment, the water on the surface of the tomographic specimen is absorbed by using water absorption filter paper.
In a preferred embodiment, the standing and air drying is to place the tomography specimen with the surface moisture absorbed dry on water-absorbing filter paper, and stand the tomography specimen in a shady and shady place for 4 to 6 hours to air dry. During the period, the water absorption filter paper is timely replaced.
In the step 4), the surface of the tomographic specimen is coated with gelatin solution, and then the tomographic specimen is kept stand and dried.
In a preferred embodiment, the concentration of the gelatin solution is 15-25%, preferably 20%.
In a preferred embodiment, the number of applications of the gelatin solution is 1 to 3, preferably 2.
The gelatin solution is obtained by hydrolyzing collagen. The collagen (called collagen for short) is the most important protein component in the skin and bone of animals. The surface of the fault specimen is coated with gelatin solution, so that the effects of isolating air and stabilizing tissue morphology can be achieved.
In a preferred embodiment, the standing and airing are standing and airing in a shady and shady place.
In the step 4), the imaging processing is to respectively scan two cross sections of the cleaned and brushed tomographic specimen by a scanner and then store the two cross sections in a hard disk of an electronic computer. As a permanent scientific research data.
In the step 4), the cleaned and brushed tomographic specimen is placed in a specimen box filled with preservation solution for permanent preservation.
In a preferred embodiment, the preservation solution is a 4-6% neutral formalin solution, preferably a 5% neutral formalin solution.
In a preferred embodiment, the cartridge is a polymethylmethacrylate plate cartridge. The specimen box is made of a transparent polymethyl methacrylate plate, a joint surface is connected by chloroform (chloroform), a small hole is drilled on the side surface of the specimen box by an electric drill to serve as a liquid injection hole for medicine injection and corrosion prevention, the cleaned and brushed fault specimen is placed in the specimen box, an upper cover is sealed, a preservation liquid is injected through the side liquid injection hole, and then the liquid injection hole is sealed and blocked for preservation. Note that the connection between the joint surface and the liquid injection hole must be tight to avoid damage and water leakage in the future.
Example 1
In a biological safety necropsy with negative pressure filtration meeting the operation requirements, hairs of corpses of patients with viral pneumonia are shaved off, dirt and oil stains on the skin of the corpses are removed, 75% ethanol water solution is used for disinfection, and then bandages are used for shaping the corpses according to anatomical postures.
And opening a curtain door of an upper opening type openable and closable structure on the side surface of one side of the pollution source chamber of the splash-proof autopsy isolating device, putting the corpse of the patient with the fixed viral pneumonia into the pollution source chamber, and closing the curtain door. An operator extends a hand into the cavity of the pollution source through the operation gloves of the sealing interfaces on the first operation port and/or the second operation port to carry out pathological dissection on the corpse. The trachea of a corpse of a patient with viral pneumonia is cut off, the thoracic cavity is cut by a T-shaped incision method, tissues outside the thoracic cavity are peeled off, ribs are exposed, costal cartilages on two sides are cut off from a second rib by a chondrotome, the distance between the tangent line and the junction of the costal cartilages and the ribs is 0.75cm, sternoclavicular joints are cut off by a scalpel, first ribs on two sides are cut off by a costal scissors, a costal arch is lifted to be tightly attached to the back of the sternum and the costal cartilages, and the sternum and the costal cartilages are removed after the diaphragm and the mediastinum are divided, so that the thoracic cavity is exposed. Then, the innominate artery and the left subclavian artery were cut off, and the trachea was cut off at a position 3.5cm above the bifurcation of the trachea at the cross section of the rib cartilage of the thoracic cage with a scalpel. Then the lung and the mediastinum are separated, the vascular nerves around the lung root are avoided, the lung root and the lung ligament are cut off vertically, and the lung is taken out. After removal of the lung, attention was paid to the location and integrity of the bronchi, pulmonary arteries, pulmonary veins, bronchial arteries and bronchopulmonary lymph nodes at the hilum of the lung. Particularly, there are phrenic nerve and pericardial phrenic vessel in front of the left lung root, and vagus nerve behind; the accessory nerve and pericardial diaphragmatic vessel are in front of the right lung root, the vagus nerve is in the back, and the strange venous arch is above the right lung root and should be avoided.
Then, the operator inserts the hand into the semi-clean chamber through the operation glove of the sealed interface on the third operation port, lifts the first isolation lifting curtain, transfers the lung taken out from the pollution source chamber into the specimen storage container in the semi-clean chamber, and closes the first isolation lifting curtain.
The lung in the specimen storage container is preserved, firstly, 10% neutral formalin solution is poured into the lung through an opening at the broken end of the trachea, namely, the trachea is cut off at a position which is about 5cm away from the bifurcation of the trachea, a plastic tube of a disposable infusion apparatus wound with gauze strips is inserted into the broken end of the trachea, the outer end of the plastic tube is aligned with the fracture of the trachea, gauze is lined between the plastic tube and the inner wall of the trachea, and then the plastic tube and the trachea are tightly tied by a suture thread around the trachea. Sucking 10% neutral formalin solution with an injector, injecting the neutral formalin solution into a person in batches slowly from the center of a plastic tube, slightly lifting the lung if gas exists in the plastic tube in the process of infusion, simultaneously pulling out the injector, slowly putting down the lung to discharge the gas, and inserting the injector into the person to infuse when the 10% neutral formalin solution is about to flow out, wherein the injection amount is determined as the injection amount. The pressure is not too high during injection, so that the tube wall is prevented from being broken. If leakage is found, the hemostatic forceps can be used for clamping or the hemostatic forceps can be used for tying.
And then soaking the lung filled with the 10% neutral formalin solution in the specimen storage container for 48 hours, taking out the lung to obtain a lung specimen, fixing the upper end (namely the trachea broken end) of the lung above the plastic barrel during soaking, and keeping the normal anatomical position. Taking out the soaked lung specimen, pouring out 0% neutral formalin solution, washing with clear water, drying the surface water of the lung specimen by using water absorption filter paper, placing the lung specimen with the surface water absorbed on the water absorption filter paper, and standing for 5 hours in a shady and shady place of the splash-proof autopsy isolation device for drying. Carefully observing the status of the lung specimen in the air-drying process, and paying attention to the fact that the lung tissue has certain humidity and cannot be too dry.
The method is characterized in that titanium dioxide powder serving as titanium dioxide dye is positioned on the surface of a lung specimen by a horizontal cross section datum line by adopting a line snapping method, and the longitudinal axis of a trachea is taken as a coronary line and the vertical line is taken as a transverse line. A cut line was drawn at a depth of 1.5cm and the lung specimen was then sealed. The operator extends the hand out of the operation glove of the sealed interface on the third operation port, lifts the second isolation lifting curtain, takes out the sealed lung specimen, and freezes at-30 ℃ for 5 days.
The operator lifts the second isolation lift curtain, puts the frozen lung specimen into the semi-clean chamber, puts the MJ345E joinery band saw into the semi-clean chamber, and closes the second isolation lift curtain. An operator stretches a hand into the semi-cleaning cavity through the operation gloves of the sealing interface on the third operation port, fixes the lung specimen on the fixed baffle of the band sawing machine, and adopts the joinery band sawing machine to saw along the sawing line to obtain the fault specimen. The saw consumption of the joinery band saw machine is less than 2mm, and the distance between the fixed baffle and the saw blade of the joinery band saw machine is equal to the thickness of the lung specimen to be sawed plus half of the saw consumption. When sawing, CT scanning is used for verification, and the thickness of the fault specimen is consistent with the scanning thickness of CT and is 0.8 mm. The length and the width of the fault specimen are carefully measured and recorded one by one, the fault specimen is placed in a sealing bag, and the serial number of the sealing bag is marked by a waterproof and oilproof pen. The operator can extend the hand out of the operation glove of the sealing interface on the third operation port, lift the second isolation lifting curtain, take out the sealed fault specimen, and sequentially place the sealed fault specimen into a medical freezing freezer at minus 30 ℃ for temporary storage.
The operator can also stretch the hand into the semi-clean chamber through the operation gloves of the sealing interface on the third operation port to continue to take pathological materials. The pathological changes of the fault specimen are observed at room temperature, and the lesion tissues of the lesion area with morphological changes are required to be picked up, wherein the pathological changes include but are not limited to color, texture, solid change, mucus change and the like. Gradient alcohol dehydration, xylene transparence, paraffin wax immersion, paraffin embedding of the examined tissue, pathological tissue section, section spreading, xylene dewaxing of the tissue section, HE staining, gradient alcohol dehydration, xylene transparence and gum sealing treatment are sequentially carried out on the examined lesion tissue, and the HE stained section is obtained and used for observing pathological histological changes under a microscope.
After the pathological section specimen is washed by water, the water on the surface of the section specimen is absorbed by water absorption filter paper, the section specimen with the absorbed water on the surface is placed on the water absorption filter paper, and the section specimen is kept stand for 4 to 6 hours in a shady and shady place to be dried. During the period, the water absorption filter paper is timely replaced. Then, the surface of the tomographic specimen was painted with 20% gelatin solution obtained by hydrolyzing collagen for 2 times, and then left to stand and air-dry in the shade of the anti-splash autopsy isolation device, and sealed with a transparent bag.
The operator stretches out the hand from the operation glove of the sealed interface on the third operation port, lifts the second isolation lifting curtain, takes out the sealed fault specimen, respectively scans the two sections of the fault specimen by the scanner, and stores the two sections in the hard disk of the electronic computer as permanent scientific research data.
And finally, lifting the second isolation lifting curtain, putting the sealed fault specimen into the semi-clean chamber, and closing the second isolation lifting curtain. An operator stretches a hand into the semi-clean chamber through an operation glove of a sealing interface on the third operation port, removes the sealing of the fault sample, puts the fault sample into a sample box made of a transparent polymethyl methacrylate plate, connects a joint surface with trichloromethane (chloroform), drills a small hole on the side surface of the sample box as a liquid injection hole by an electric drill, puts the fault sample into the sample box, adds a sealing upper cover, injects 5% neutral formalin solution through the side liquid injection hole, then seals and stores the liquid injection hole. Note that the connection between the joint surface and the liquid injection hole must be tight to avoid damage and water leakage in the future. The operator stretches out the hand from the operation glove of the sealing interface on the third operation port, lifts the second isolation lifting curtain, and transfers the specimen box filled with the fault specimen to a place for long-term storage. In the process, the splash-proof autopsy isolation device ensures that the pollution source does not directly contact with an operator.
As shown in figure 2, the tomographic specimen prepared by the invention has clear anatomical structure and high lesion display degree, and can well meet the general observation requirement.
The tomographic specimen prepared in the invention can be verified by its HE stained section:
as shown in FIG. 3, the lung interstitial congestion and edema, lymphocyte and monocyte infiltration in the HE stained section of the sectional specimen result in the widening of part of the alveolar space, no exudate or only a small amount of serous fluid is seen in the alveolar cavity, which indicates that the sectional specimen is microscopically consistent with interstitial pneumonia and belongs to the lung of patients with interstitial pneumonia of patients with viral pneumonia.
As shown in fig. 4, in addition to the pulmonary interstitium, lesions in HE stained sections of the tomographic specimens may spread to the alveolar space, where different amounts of serous fluid, cellulose, monocytes, macrophages, etc. appear. It is indicated that the tomographic specimen shows particularly serious viral pneumonia under the microscope. In the HE stained section of the tomographic specimen of the patient with viral pneumonia, the serous cellulose exudate is concentrated to form a layer of uniform red-stained membranous material, namely a transparent membrane, on the alveolar cavity surface.
Therefore, the HE stained section of the tomographic specimen prepared in the invention has clear images and can perfectly show the viral pneumonia lung tissue images as shown in FIGS. 3 and 4.
In conclusion, the preparation method of the fault specimen based on the lung of the viral pneumonia patient provided by the invention reserves precious clinical pathological data for the subsequent study of the viral pneumonia, and avoids unnecessary infection of an operator in the preparation process. Therefore, the invention effectively overcomes various defects in the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be accomplished by those skilled in the art without departing from the spirit and scope of the present invention as set forth in the appended claims.
Claims (10)
1. A preparation method of a lung fault specimen, which is to place a corpse in a splash-proof autopsy isolation device for processing in a biological safe autopsy room, comprises the following steps:
1) dissecting the corpse, taking out the lung, soaking the lung in a fixing solution, and taking out the lung to obtain a lung specimen;
2) performing horizontal cross section datum line positioning on the surface of the lung specimen obtained in the step 1) by adopting a line snapping method, then scribing a saw line, and freezing the lung specimen;
3) sawing the lung specimen frozen in the step 2) along a sawing line, and verifying by CT scanning to obtain a fault specimen;
4) carrying out pathological material taking, cleaning and glue brushing on the fault specimen obtained in the step 3), then carrying out imaging treatment, and sealing and storing.
2. The method for preparing a lung tomographic specimen as recited in claim 1, wherein the corpse is disposed in a splash-proof autopsy isolation device for processing, comprising the steps of:
a) the splash-proof autopsy isolation device comprises a pollution source chamber and a semi-clean chamber which are isolated by an isolation surface, a curtain door on one side surface adjacent to the isolation surface on the pollution source chamber is opened, a corpse is placed in the pollution source chamber, and after the curtain door is closed, the corpse is dissected through at least one first operation port arranged on the other side surface adjacent to the isolation surface and/or at least one second operation port arranged on the side surface opposite to the isolation surface, and the lung is taken out;
b) a second isolation lifting curtain is arranged on the side surface, opposite to the isolation surface, of the semi-cleaning chamber, and the lung is transferred to the semi-cleaning chamber from the pollution source chamber through the first isolation lifting curtain arranged on the isolation surface through at least one third operation port arranged on the second isolation lifting curtain;
c) placing the lung in a specimen storage container arranged in the semi-cleaning chamber, soaking the lung in a fixing solution to obtain a lung specimen, scribing and sawing the lung specimen, opening a second isolation lifting curtain, and taking out the lung specimen for freezing;
d) placing the frozen lung specimen into a semi-clean chamber through a second isolation lifting curtain, performing saw cutting through a third operation port to obtain a fault specimen, performing pathological material taking, cleaning and glue brushing on the fault specimen, sealing, and taking out the fault specimen through the second isolation lifting curtain for imaging treatment;
e) and putting the imaged fault sample into a semi-clean chamber through a second isolation lifting curtain to manufacture a specimen box, and taking out the specimen box.
3. The method for preparing the tomographic specimen of the lung according to claim 1, wherein the anti-splash autopsy isolation apparatus comprises a contamination source chamber and a semi-clean chamber which are adjacent to each other, the contamination source chamber and the semi-clean chamber are isolated by an isolation surface, and the isolation surface is provided with a first isolation lifting curtain; the side surface of one side of the pollution source chamber adjacent to the isolation surface is a curtain door, the side surface of the other side of the pollution source chamber adjacent to the isolation surface is provided with at least one first operation port, and the side surface opposite to the isolation surface is provided with at least one second operation port; and a second isolation lifting curtain is arranged on the side surface of the semi-cleaning chamber opposite to the isolation surface, and at least one third operation port is arranged on the second isolation lifting curtain.
4. The method of claim 1, wherein the cadaver is removed by shaving hair, removing dirt and oil from the skin of the cadaver, and then sterilizing the cadaver with a disinfectant, and then fixing the cadaver with a bandage; the disinfectant is 70-80% ethanol water solution by volume percentage.
5. The method for preparing the lung tomographic specimen as recited in claim 1, wherein in the step 1), the lung is immersed in the fixative by injecting the fixative into the lung through the opening of the severed end of the trachea, and then immersing the lung filled with the fixative in the fixative; the fixing solution is 5-15% of neutral formalin solution; the soaking time of the lung in the fixing solution is 46-50 h.
6. The method for preparing a lung tomographic specimen according to claim 1, wherein the step 2) includes any one or more of the following conditions:
A1) the titanium dioxide dye is titanium dioxide;
A2) the scribing depth of the sawing line is 1-2 cm;
A3) the lung specimen is sealed before freezing, wherein the freezing is to freeze the lung specimen at-30 ℃ for 4-6 days.
7. The method for preparing the lung tomographic specimen as recited in claim 1, wherein in the step 3), the sawing is performed by a band saw machine, and a saw consumption of the band saw machine is less than 2 mm.
8. The method for preparing the lung sectional specimen according to claim 1, wherein the pathological sampling comprises the following steps:
A) observing pathological changes of the fault specimen at room temperature, and taking lesion tissues for examination of a morphologically changed lesion area;
B) sequentially carrying out gradient alcohol dehydration, xylene transparence, paraffin wax immersion, paraffin embedding of the detected tissue, pathological tissue section, slide sticking, xylene dewaxing of the tissue section, HE dyeing, gradient alcohol dehydration, xylene transparence and gum sealing treatment on the detected lesion tissue to obtain an HE dyed section.
9. The method for preparing the lung fault specimen according to claim 1, wherein the brushing is performed by brushing the surface of the fault specimen with gelatin solution, standing and drying; the gelatin solution is obtained by hydrolyzing collagen, the concentration of the gelatin solution is 15-25%, and the brushing frequency of the gelatin solution is 1-3 times.
10. Use of the method of preparing a lung sectional specimen according to any one of claims 1 to 9 for preparing a lung sectional specimen of a patient with viral pneumonia.
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