CN113063751B - Forensic medicine pulmonary fat embolism analysis method based on infrared spectrum imaging technology - Google Patents
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Abstract
The invention provides a forensic medicine pulmonary fat embolism analysis method based on infrared spectrum imaging technology, which comprises the following steps: s1, obtaining a biological sample, preprocessing the biological sample, and putting the biological sample into an infrared spectrum detection table; s2, collecting chemical information and analyzing chemical components of the biological sample by using an infrared spectrum detection platform to obtain a lipid content pseudo-color image; s3, constructing an artificial intelligent neural network model, processing the pseudo-color image by adopting the artificial intelligent neural network model to obtain the number of lipid pixels in blood vessels, and performing pulmonary tissue embolism analysis according to the number of lipid pixels in the blood vessels. The invention positions lipid components in alveolar wall capillaries and interstitial small blood vessels according to the characteristic functional group vibration mode, and realizes the death cause identification of the forensic pulmonary fat embolism by qualitative and quantitative analysis of the lipid components.
Description
Technical Field
The invention relates to the technical field of forensic pathology diagnosis, in particular to a forensic pulmonary fat embolism analysis method based on infrared spectrum imaging technology.
Background
Pulmonary Fat Embolism (PFE) is a phenomenon of lipid droplet embolism in alveolar wall capillaries and interstitial small blood vessels which mainly occurs after serious trauma, especially long bone fracture and fat tissue-rich soft tissue contusion and pressure injury. The main death mechanism is that after trauma, the lipid drop component enters into the systemic circulation and reaches the blood vessel of the lung through the right heart, so that the blood-gas exchange dysfunction is caused, and the human body is suffocated to die. In forensic practice, PFE can be used as both a primary cause of death and an auxiliary cause of death for assessing the degree of involvement of trauma in death; also, PFE may be considered a life response, suggesting that the victim remains alive at or shortly after the time of trauma.
Diagnosis of PFE relies primarily on traditional histopathological methods, including conventional HE staining and associated lipid-specific staining (e.g., sudan III, sudan IV, sudan black B, and oil red-O). In HE staining, lipid droplet components in pulmonary vessels are often dissolved by organic solvents, showing a beaded-like change in morphology, requiring further special staining for evidence. However, the related special dyeing method has the following problems: (1) the dyeing process is time consuming, labor intensive and produces a large amount of toxic waste; (2) the quality of dyeing is usually determined by the proficiency of technicians, the dyeing effect among different experiment platforms has larger difference, and the repeatability of results is poor; (3) in the process of staining and washing the slide, because the lipid drop is loosely combined with the surrounding tube cavity, the phenomenon that the lipid drop is deviated or diffused to the outside of the blood vessel is easily caused, thereby causing the occurrence of false negative results; (4) specially stained sections are relatively difficult to preserve, fade over time, and retrospective inspection is difficult to perform. Therefore, there is an urgent need to develop an objective, environmentally friendly, reproducible method for the in-depth diagnosis and study of PFE.
Disclosure of Invention
Aiming at the technical problems, the invention provides a forensic medicine pulmonary fat embolism analysis method based on an infrared spectrum imaging technology, which is used as a label-free and nondestructive spectrum detection technology, and an FTIR spectrum can show characteristic peaks corresponding to specific chemical functional groups in a sample. And (3) positioning lipid components in alveolar wall capillaries and interstitial small blood vessels according to the characteristic functional group vibration mode, and performing qualitative and quantitative analysis on the lipid components. The identification of the death cause of the forensic pulmonary fat embolism is realized.
In order to achieve the purpose, the invention provides the following scheme: the invention provides a forensic medicine pulmonary fat embolism analysis method based on infrared spectrum imaging technology, which comprises the following steps:
s1, obtaining a biological sample, preprocessing the biological sample, and putting the processed biological sample into an infrared spectrum detection table;
s2, collecting chemical information and analyzing chemical components of the biological sample by using an infrared spectrum detection platform to obtain a lipid content pseudo-color image;
s3, constructing an artificial intelligent neural network model, processing the lipid content pseudo-color image by adopting the artificial intelligent neural network model to obtain the number of lipid pixels in blood vessels, and carrying out pulmonary fat embolism analysis according to the number of lipid pixels in the blood vessels.
Preferably, the process of step S1 is:
s1.1, placing a biological sample into 4% paraformaldehyde fixing solution for fixing for 24 hours, performing gradient dehydration on the sample after the sample is fixed for 24 hours by using 10%, 20% and 30% sucrose, cleaning and slicing;
s1.2, preparing the sliced biological sample into a slice, placing the slice on a calcium fluoride or barium fluoride slice, and placing the slice on an infrared spectrum detection table after the tissue is completely dried.
Preferably, the process of step S2 is:
s2.1, debugging parameters of an infrared spectrum detection table, and defining a sample scanning area on the detection table;
and S2.2, based on the sample scanning area, adopting an infrared spectrum detection table to collect macromolecular substance chemical information of the biological sample and analyzing chemical components to obtain a lipid content pseudo-color image.
Preferably, the parameters of the infrared spectroscopy detection station include grating size, spectral resolution and focal plane array detector size.
Preferably, the grating size is 2.5 μm × 2.5 μm; the spectral resolution is 4cm-1(ii) a The focal plane array detector is 64 x 64 in size and one level of zero padding.
Preferably, the chemical composition analysis process is: 1725 + 1745cm in the spectrum-1And 2800--1And performing integration treatment on the two wave bands, and constructing a pseudo-color image according to an integration quantitative result, wherein the larger the integration value is, the higher the lipid content of the corresponding region is.
Preferably, the process of step S3 is:
s3.1, carrying out image turnover, scale transformation, contrast transformation and normalization processing on the lipid content pseudo-color image;
s3.2, constructing an artificial intelligent neural network model, and training the artificial intelligent neural network model by adopting the processed data of the lipid content pseudo-color image;
and S3.3, marking the intravascular lipid in the lipid content pseudo-color image by adopting the trained artificial intelligent neural network model, carrying out quantitative analysis according to the pixel number of the intravascular lipid, and determining that the pulmonary fat embolism is dead when the lipid number marked in the blood vessel is more than a specific value.
Preferably, when the intravascular lipid is labeled more than 30%, it is considered to be pulmonary fat embolism death.
The invention discloses the following technical effects:
the invention can position lipid components in alveolar wall capillaries and interstitial small vessels according to the characteristic functional group vibration mode, and realizes the death cause identification of the forensic pulmonary fat embolism by qualitative and quantitative analysis of the lipid components; the analysis process of the invention does not need any chemical reagent, thus realizing the detection purposes of green, environmental protection and no pollution; meanwhile, the obtained result highly depends on the parameter setting of the infrared spectrum imager, the operation is simple, convenient and easy, and the chemical analysis with high efficiency, rapidness and high repeatability can be carried out by combining the strong area data acquisition capacity of the focal plane array detector; because the method belongs to nondestructive and in-situ detection, and fat components in blood vessels cannot shift or leak, the probability of false negative results is far lower than that of the traditional special dyeing method; more importantly, the method provided by the invention is different from the traditional special staining storage mode in that the slice data is stored in a digital mode, and has the advantages of high stability and strong backtracking property.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.
FIG. 1 is a schematic flow diagram of the process of the present invention;
FIG. 2 is a schematic view of spectral data collected by an IR imager in accordance with an embodiment of the invention;
fig. 3 is a pseudo-color drawing according to an embodiment of the present invention, in which: FIG. 3(a) is 2800--1Infrared imaging schematic diagram of (1); FIG. 3(b) is 1725-1745cm-1Schematic diagram of infrared imaging.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.
As shown in fig. 1, the invention provides a forensic lung fat embolism analysis method based on infrared spectrum imaging technology, comprising the following steps:
s1, tissue sample treatment: obtaining a biological sample, preprocessing the biological sample, making the preprocessed biological sample into a 4-10 mu m thick sheet, placing the sheet on a calcium fluoride or barium fluoride slice, and placing the sheet on an infrared spectrum detection table after the tissue is completely dried.
The sample can be pretreated by the following three methods before being made into a thin sheet: 1. directly slicing blood stains on the surface of a sample after cleaning; 2. fixing the sample in 4% paraformaldehyde fixing solution for 24 hours, and slicing; 3. the samples after 24 hours of fixation were dehydrated with a 10%, 20%, 30% sucrose gradient, washed and sliced.
S2, chemical information acquisition and chemical component analysis: debugging related parameters of the ultra-high resolution Fourier transform infrared spectrum imager, determining a sample scanning area on a detection table, executing macromolecular substance chemical information acquisition work, and determining the chemical distribution condition of lipid components in a tissue sample slice according to related spectrum chemical absorption peaks.
The instrument debugging parameters include: the grating size is 2.5 μm × 2.5 μm, and the spectral resolution is 4cm-1Focal plane array detector size 64 x 64, first order zero fill. The infrared spectrum adopts a transmission mode, each sample is scanned for 64 times and then averaged, and the transmission infrared spectrum is recorded at 800-4000cm-1In between, a new background spectrum is recorded every 15 sample spectra.
Processing the obtained infrared spectrum data: atmosphere Compensation (CO) of the obtained infrared spectrum2Compensation) and then baseline correction is carried out, and a Rubbbeband correction method is selected for correction. Vector normalization is then performed and the spectrum is smoothed with the smoothing factor set to 9.
The spectral data collected by the ir imager of this embodiment is shown in fig. 2.
Chemical component analysis: 1725 + 1745cm in the spectrum-1(C ═ O bond of fatty acid ester) and 2800-3000cm-1(C-H、C-H2、C-H3Lipid region), and the integrated quantitative result is presented in the form of a pseudo-color image, the larger the integrated value is, the higher the lipid content of the corresponding region is, and the pseudo-color image of the integrated quantitative result obtained in this embodiment is specifically shown in fig. 3.
S3, artificial intelligence technology diagnosis: the ratio of the blood vessel lipid component in the scanning area is quantitatively and qualitatively analyzed by adopting a related artificial intelligence technology, so that the diagnosis of the pulmonary fat embolism and death is determined.
Firstly, an artificial intelligent neural network model is constructed and trained, wherein the artificial intelligent model comprises RCNN, Yolo, U-Net and the like. The training data is the lipid distribution pseudo-color map obtained in step S2 and the binary image corresponding to the lipid component distribution. The selected pictures are all artificially filtered and the filtered pictures are cropped to squares. Because, there are fewer samples of fat emboli. Therefore, in this embodiment, the picture is randomly preprocessed, including flipping, scale transformation, and contrast transformation, and all pictures are normalized. The pre-processed data was randomly divided into 4 sets, only one of which was used as a test set for evaluation of the composite image. The remaining 3 data were used as a training set to generate a composite image. And finally, putting the synthesized image as a training set into the constructed artificial intelligent neural network model for training.
The trained artificial intelligence model can label the intravascular lipid in the chemical information map acquired in step S2 and perform quantitative analysis according to the number of pixels, and when the proportion of lipid pixels is greater than 30%, the lipid is considered as pulmonary fat embolism death.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (7)
1. A forensic medicine pulmonary fat embolism analysis method based on infrared spectrum imaging technology is characterized by comprising the following steps:
s1, obtaining a biological sample, preprocessing the biological sample, and putting the processed biological sample into an infrared spectrum detection table;
s2, collecting chemical information and analyzing chemical components of the biological sample by using an infrared spectrum detection platform to obtain a lipid content pseudo-color image;
s3, constructing an artificial intelligent neural network model, processing the lipid content pseudo-color image by adopting the artificial intelligent neural network model to obtain the number of lipid pixels in blood vessels, and performing pulmonary tissue embolism analysis according to the number of lipid pixels in the blood vessels;
the process of S3 is:
s3.1, carrying out image turnover, scale transformation, contrast transformation and normalization processing on the lipid content pseudo-color image;
s3.2, constructing an artificial intelligent neural network model, and training the artificial intelligent neural network model by adopting the processed data of the lipid content pseudo-color image;
and S3.3, marking the intravascular lipid in the lipid content pseudo-color image by adopting the trained artificial intelligent neural network model, carrying out quantitative analysis according to the pixel number of the intravascular lipid, and determining that the pulmonary fat embolism is dead when the lipid number marked in the blood vessel is more than a specific value.
2. The forensic lung fat embolism analysis method based on infrared spectroscopy imaging technology according to claim 1 wherein the process of S1 is:
s1.1, placing a biological sample into 4% paraformaldehyde fixing solution for fixing for 24 hours, performing gradient dehydration on the sample after the sample is fixed for 24 hours by using 10%, 20% and 30% sucrose, cleaning and slicing;
s1.2, preparing the sliced biological sample into a slice, placing the slice on a calcium fluoride or barium fluoride slice, and placing the slice on an infrared spectrum detection table after the tissue is completely dried.
3. The forensic lung fat embolism analysis method based on infrared spectroscopy imaging technology according to claim 1 wherein the process of S2 is:
s2.1, debugging parameters of an infrared spectrum detection table, and defining a sample scanning area on the detection table;
and S2.2, based on the sample scanning area, adopting an infrared spectrum detection table to collect macromolecular substance chemical information of the biological sample and analyzing chemical components to obtain a lipid content pseudo-color image.
4. The forensic lung fat embolism analysis method according to claim 3 based on infrared spectroscopy imaging technology wherein the parameters of the infrared spectroscopy detection station include grating size, spectral resolution and focal plane array detector size.
5. The forensic medical pulmonary fat embolism analysis method based on infrared spectroscopy imaging technology according to claim 4 wherein the grating size is 2.5 μ ι η x 2.5 μ ι η; the spectral resolution is 4cm-1(ii) a The focal plane array detector is 64 x 64 in size and one level of zero padding.
6. The forensic medicine pulmonary fat embolism analysis method based on infrared spectroscopy imaging technology according to claim 3 wherein the chemical composition analysis process is: 1725 + 1745cm in the spectrum-1And 2800--1And performing integration treatment on the two wave bands, and constructing a pseudo-color image according to an integration quantitative result, wherein the larger the integration value is, the higher the lipid content of the corresponding region is.
7. The forensic lung fat embolism analysis method based on infrared spectroscopy imaging technology as claimed in claim 1 wherein lung fat embolism is considered dead when intravascular lipids are labelled more than 30%.
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