CN110042141A - A kind of diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging - Google Patents
A kind of diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging Download PDFInfo
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- 238000007689 inspection Methods 0.000 title claims abstract description 38
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- 238000000120 microwave digestion Methods 0.000 claims abstract description 15
- 238000012545 processing Methods 0.000 claims abstract description 9
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 28
- 230000029087 digestion Effects 0.000 claims description 22
- 239000012496 blank sample Substances 0.000 claims description 15
- 239000013068 control sample Substances 0.000 claims description 11
- 239000002253 acid Substances 0.000 claims description 10
- 210000000056 organ Anatomy 0.000 claims description 10
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 10
- 239000012498 ultrapure water Substances 0.000 claims description 10
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- 238000002604 ultrasonography Methods 0.000 claims description 3
- 239000005446 dissolved organic matter Substances 0.000 claims description 2
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims 3
- 238000004364 calculation method Methods 0.000 claims 1
- 238000004458 analytical method Methods 0.000 description 4
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- 210000004072 lung Anatomy 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 241000206761 Bacillariophyta Species 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
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- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 2
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Abstract
The invention discloses a kind of diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, it include: sample after micro-wave digestion, pH are adjusted, enter a kind of Flow Optical microscopic imaging device with the type of flow, particle therein is quickly shot using the micro- camera of high speed, and automate identification software using diatom and automatically identify diatom from particle photo and classify, to making survey report after data processing.This method significantly improves checkability and accuracy, reduces labor intensity, has a good application prospect in medicolegal practice.
Description
Technical field
The present invention relates to medicolegal examination field, in particular to a kind of medical jurisprudence diatom based on Flow Optical micro-imaging
The method of inspection.
Background technique
The analysis on cause of death of corpse is the great difficult problem in medical jurisprudence in water.Although scholars are drowned diagnosis to diatom examination
Value there are still disagreements, but its had always been considered as since 1904 by invention be it is a kind of be more reliably drowned diagnostic means,
Particularly with the identification of the decomposed body cause of the death in water, it is considered as the best approach (" goldstandard ").Before diatom examination includes sample
Reason and analysis (i.e. qualitative analysis) and the analysis of content (i.e. quantitative analysis) to diatom type contained by sample.Diatom examination practice
In, sample pre-treatments mainly use strong acid to heat resolution-centrifugal process, research shows that the method has resolution low efficiency, resolution effect
The notable defects such as low, the pollution environment of difference, the diatom rate of recovery；Diatom floristic analysing need to apply diatom taxology professional knowledge, usually
Legal medical expert worker asks diatom to learn expert assistance after shooting diatom photo and carries out diatom Identification of Species；And traditional diatom content point
Analysis method is: the visual field is searched, takes pictures, counts and is counted one by one manually under an optical microscope, though legal medical expert worker can
Complete independently, but low efficiency, working strength are high, examine error big.For the above-mentioned deficiency of traditional diatom examination, with the present inventor
In doctor Hu Sunlin be that " micro-wave digestion-vacuum filtration-scanning electron microscope diatom examination side has been invented by the main seminar for completing people
(patent of invention number: ZL 2,009 1 0214550. 2, ZL 2,010 1 0115728. is 0) for method ".This method is that diatom examination is ground
An important breakthrough for studying carefully field is that of obtaining national science and technology progress second prize project " legal medical expert's diatom examination key skill in 2017
The core achievement of art and equipment ", relative to conventional method, this method is improving inspection sensitivity, anti-pollution, high efficiency, safety
Property etc. all achieve marked improvement, in particular, will be drowned corpse diatom examination positive rate by 28% improve to 97.4%, mention significantly
High diatom examination is drowned diagnostic value.
However, " micro-wave digestion-vacuum filtration-scanning electron microscope methods for detecting diatom " still carries out diatom knowledge using manual type
Other and Identification of Species, furthermore the image photographic speed of scanning electron microscope constrains the raising of checkability not as good as light microscopic, these factors.
Summary of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of high degree of automations, efficient, accurate method
Medicine methods for detecting diatom.
In order to achieve the above technical purposes, the invention proposes a kind of medical jurisprudence diatoms based on Flow Optical micro-imaging
The method of inspection, this method significantly improve checkability and accuracy, reduce labor intensity, have in medicolegal practice good
Good application prospect, specific as follows:
A kind of diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, successively to the sample in medical jurisprudence diatom examination
Carry out following examine:
(1) micro-wave digestion, makes sample under microwave irradiation effect and strong acid chemically reacts, with quick dissolved organic matter；It is micro-
The purpose of wave resolution is the frustule wall for extracting acid corrosion-resistant from sample, can be used as being drowned indicant.
(2) digestion solution pH is adjusted, and carries out pH adjusting to the liquid obtained after micro-wave digestion；
(3) the particle dynamic optical micro-imaging in digestion solution, the digestion solution after adjusting pH enter Flow Optical with the type of flow
Microscopic imaging device, particle (including frustule wall) therein after testing window when by the micro- camera snapshots of high speed；
(4) diatom is identified automatically, using diatom automatic identifying system to Granule Micrograph that is captured, being stored in computer into
Row diatom automatic identification and classification；
(5) data processing and survey report production, handle inspection data and are analyzed, obtain in sample the type of diatom and
Content；Then the type and content for comparing diatom contained by each sample, judges matching degree；Survey report is made, finally to be drowned
Diagnosis provides foundation.
Preferably, removing internal organs group in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging
It knits with outside water sample inspection, is additionally provided with blank sample and control sample survey, occurs to avoid false positive and false negative result.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, the blank
Sample is the sample without diatom, such as pure water；The control sample is the standard sample containing center guiding principle and pennates diatom,
Diatom concentration is 1-100/mL, and size is 1-100 μm.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, step (1) institute
It states micro-wave digestion to include the following steps: to add to sample in micro-wave diminishing pot, concentrated nitric acid (and/or other strong acid) microwave afterwards is added
Resolution, digestion solution pure water or ultrapure water dilution；Digestion solution is transferred to test tube, it is spare；Because the counteracting tank after resolution may be residual
It stays diatom, pollutes next group sample, to remove diatom residue, counteracting tank is placed in 1-5NNaOH solution and is impregnated, then with surpassing
Pure water cleaning；Or HF solution (or other can dissolve the acid of diatom) to counteracting tank is added to carry out micro-wave digestion to dissolve frustule
Wall.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, step (1) institute
Stating sample includes 1-10g organs and tissues (lung, liver, kidney, marrow etc.), 1-10mL water sample, 1-10mL blank sample and 1-
10mL controls sample.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, step (2) institute
State digestion solution pH adjustment step specifically:
(1) it is filtered by vacuum: using Vacuum filtration device, the digestion solution that step (1) is obtained filters, and adds pure water or ultrapure after having filtered
Water continues to filter, and makes filter membrane surface close to neutrality；Parameter is filtered by vacuum are as follows: vacuum degree 100-700mmHg, used filter membrane
Diameter 10-50mm, aperture are 0.1-3.0 μm；
(2) particle is desorbed: the filter membrane handled using the method for ultrasound desorption or vacuum suction makes the particle of its surface enrichment
Desorption is into pure water or ultrapure water.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, step (3) institute
The particle dynamic optical micro-imaging in digestion solution is stated to include the following steps: suspension obtained by step (2) adding to Flow Optical
In the sample injector of microscopic imaging device, suspension is transported to detection system with the flow velocity of 0.01mL/min ~ 10mL/min, when
Grain object (including frustule wall) after testing window when, image is shot by the micro- camera of high speed, amplification factor 40-
2000X。
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, the fluid
Optical micro imaging device includes: sample injector, fluid delivery system, light source, detection window, the micro- camera of high speed, computer and control
Software processed.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, step (4) institute
Stating the process that diatom is identified automatically includes: to identify software to captured automatically using the diatom for including image recognition and calssification technology
Image is handled and is analyzed in real time, is identified diatom and is classified, and diatom qualification result automatically to be stored in computer.
Preferably, in the above-mentioned diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, step (5) institute
It states data processing and survey report production includes the following steps:
(1) statistical check data；
(2) only when blank sample inspection result is negative, control sample detection diatom type is identical as Known Species, diatom content is missed
When difference is less than given value 10-20%, sample inspection result is significant, can provide survey report；
(3) survey report is made, by Judicial Identification Document related request production survey report, in its check conclusion part,
The type and content that diatom in sample should be stated should also state organs and tissues and water sample if sample includes organs and tissues and water sample
The matching degree of diatom examination result.
Compared with prior art, the invention has the following beneficial effects:
1. efficiently: the present invention improves checkability by two kinds of technologies: first, scanning electron microscope is replaced using the micro- camera of high speed,
Substantially increase particle image taking speed；Second, artificial diatom is replaced to identify mode, automatic diatom mirror in a manner of the identification of automatic diatom
Determine system to automatically analyze to taking a picture in real time, identify diatom and classify, avoids traditional methods for detecting diatom and " microwave disappears
Solution-vacuum filtration-scanning electron microscope methods for detecting diatom " time-consuming, the deficiency of effort.
2. accurate: compared with conventional method and " micro-wave digestion-vacuum filtration-scanning electron microscope methods for detecting diatom ", this hair
It is bright that diatom identification is carried out using diatom automatic identifying system, reviewer is overcome caused by diatom identification level difference
Personal error.
3. high degree of automation, labor intensity are low: with conventional method and " micro-wave digestion-vacuum filtration-scanning electron microscope diatom
The method of inspection " is compared, and the present invention uses automatic sampling and automatic detection mode, is avoided search, label, is amplified, takes pictures, identifying
Etc. manual steps, greatly reduce the labor intensity of reviewer.
Detailed description of the invention
Fig. 1 is Flow Optical microscopic imaging device schematic illustration；
Fig. 2 is the diatom picture shot by Flow Optical microscopic imaging device；
(a) the diatom photo quickly shot, 100X；
(b) the diatom photo of time lapse, 200X；
Fig. 3 is inspection process figure of the invention.
Specific embodiment
Embodiment 1:
The present invention include the following steps: micro-wave digestion, digestion solution pH adjusting, the particle dynamic optical micro-imaging in digestion solution,
Diatom is identified automatically, data processing and survey report make.
1 sample
Including sample, blank sample (I and II), control sample.
1.1 sample
Organs and tissues (lung, liver, kidney, marrow etc.) and water sample.
1.2 blank sample I
Pure water or ultrapure water.
1.3 blank sample II
Pure water or ultrapure water.
1.4 control samples
Standard sample containing center guiding principle and pennates diatom, diatom concentration are 1-100/mL, and size is 1-100 μm.
2 checkout sequences
By blank sample I, control sample, blank sample II, the sequence for closing organ-tissue (liver, kidney, marrow etc.), lung, water sample
Successively test.
3 methods of inspection
All samples are examined by the following method.
3.1 micro-wave digestions:
Sample is added in micro-wave diminishing pot, concentrated nitric acid (and/or other acid) micro-wave digestion afterwards, digestion solution pure water or super is added
Pure water dilution；Digestion solution is transferred to test tube, it is spare.Because the counteracting tank after resolution may remain diatom, next group sample is polluted,
To remove diatom residue, counteracting tank is placed in 1-5NNaOH solution and is impregnated, then is cleaned with ultrapure water；Or add HF solution (or
Other can dissolve diatoms acid) to counteracting tank carry out micro-wave digestion to dissolve frustule wall.
Amount of samples is as follows:
Blank sample (I and II): 1-10mL
Control sample: 1-10mL
Organs and tissues (lung, liver, kidney, marrow etc.): 1-10g
Water sample: 1-10mL
Micro-wave digestion condition setting: by microwave dissolver manufacturer's recommended parameter or the parameter setting after optimum experimental.
3.2 digestion solution pH are adjusted
Digestion solution pH adjustment step specifically:
3.2a vacuum filtration
Using Vacuum filtration device, the digestion solution that 3.1 processing obtain is filtered, adds pure water or ultrapure water to continue to filter after having filtered,
Make filter membrane surface close to neutrality；
Parameter is filtered by vacuum are as follows: vacuum degree 100-700mmHg, used filter membrane diameter 10-50mm, aperture 0.1-3.0
µm。
3.2b particle desorption
The filter membrane obtained using the method processing 3.2a of ultrasound desorption or vacuum suction, is desorbed the particle of its surface enrichment to 1-
In 50mL pure water or ultrapure water.
4 particle dynamic optical micro-imagings
The suspension that 3.2b is obtained adds in the sample injector of Flow Optical microscopic imaging device, suspension with 0.01mL/min ~
The flow velocity of 10mL/min is transported to detection system, and when particulate matter (including frustule wall) window after testing, image is
By the micro- camera shooting of high speed, amplification factor 40-2000X.
5 diatoms are identified automatically
Using the diatom based on the image recognition and calssifications Technology design such as artificial intelligence, machine vision, deep learning, neural network
Automatic identification software is handled and is analyzed in real time to image captured by 4, is identified diatom and is classified, diatom qualification result is automatic
It is stored in computer.
6 data processings and survey report production
6a statistical check data.
6b is only negative when blank sample inspection result, and control sample detection diatom type is identical as Known Species, diatom contains
When measuring error less than given value 10-20%, sample inspection result is significant, can provide survey report.
6c makes survey report.Survey report is made by Judicial Identification Document production related request, examines and ties at it
By part, the type and content of diatom in sample should be stated, if sample includes organs and tissues and water sample, should also state organs and tissues
With the matching degree of water sample diatom examination result.
Claims (10)
1. a kind of diatom inspection method in medical jurisprudence based on Flow Optical micro-imaging, it is characterised in that medical jurisprudence diatom examination
In sample successively carry out following examine:
(1) micro-wave digestion, makes sample under microwave irradiation effect and strong acid chemically reacts, with quick dissolved organic matter；
(2) digestion solution pH is adjusted, and carries out pH adjusting to the liquid obtained after micro-wave digestion；
(3) the particle dynamic optical micro-imaging in digestion solution, the digestion solution after adjusting pH enter Flow Optical with the type of flow
Microscopic imaging device, particle therein after testing window when by the micro- camera snapshots of high speed；
(4) diatom is identified automatically, using diatom automatic identifying system to Granule Micrograph that is captured, being stored in computer into
Row diatom automatic identification and classification；
(5) data processing and survey report production, handle inspection data and are analyzed, obtain in sample the type of diatom and
Content；Then the type and content for comparing diatom contained by each sample, judges matching degree；Survey report is made, finally to be drowned
Diagnosis provides foundation.
2. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, which is characterized in that
The sample includes organs and tissues, water sample, blank sample and control sample.
3. the diatom inspection method in medical jurisprudence according to claim 2 based on Flow Optical micro-imaging, which is characterized in that
The blank sample is the sample without diatom；The control sample is the standard sample containing center guiding principle and pennates diatom,
Its diatom concentration is 1-100/mL, and size is 1-100 μm.
4. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, which is characterized in that
Step (1) described micro-wave digestion includes the following steps: to add to sample in micro-wave diminishing pot, and micro-wave digestion after strong acid is added, disappears
Solve liquid pure water or ultrapure water dilution；Digestion solution is transferred to test tube, it is spare；Counteracting tank is placed in NaOH solution and is impregnated, then
It is cleaned with ultrapure water；Or HF solution or other acid to counteracting tanks that can dissolve diatom is added to carry out micro-wave digestion to dissolve frustule
Wall.
5. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, which is characterized in that
Step (1) described sample includes 1-10g organs and tissues, 1-10mL water sample, 1-10mL blank sample and 1-10mL control sample
Product.
6. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, which is characterized in that
Step (2) the digestion solution pH adjustment step specifically:
(1) it is filtered by vacuum: using Vacuum filtration device, the digestion solution that step (1) is obtained filters, and adds pure water or ultrapure after having filtered
Water continues to filter, and makes filter membrane surface close to neutrality；Parameter is filtered by vacuum are as follows: vacuum degree 100-700mmHg, used filter membrane
Diameter 10-50mm, aperture are 0.1-3.0 μm；
(2) particle is desorbed: the filter membrane handled using the method for ultrasound desorption or vacuum suction makes the particle of its surface enrichment
Desorption is into pure water or ultrapure water.
7. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, which is characterized in that
Particle dynamic optical micro-imaging in step (3) described digestion solution includes the following steps: to add to suspension obtained by step (2)
In the sample injector of Flow Optical microscopic imaging device, suspension is transported to detection with the flow velocity of 0.01mL/min ~ 10mL/min
System, when particulate matter window after testing, image is shot by the micro- camera of high speed, amplification factor 40-2000X.
8. the diatom inspection method in medical jurisprudence according to claim 7 based on Flow Optical micro-imaging, which is characterized in that
The Flow Optical microscopic imaging device includes: sample injector, fluid delivery system, light source, detection window, the micro- camera of high speed, meter
Calculation machine and control software.
9. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, which is characterized in that
The process that step (4) described diatom is identified automatically include: using include image recognition and calssification technology diatom identify automatically it is soft
Part is handled and is analyzed in real time to captured image, is identified diatom and is classified, and diatom qualification result automatically to be stored in calculating
Machine.
10. the diatom inspection method in medical jurisprudence according to claim 1 based on Flow Optical micro-imaging, feature exist
In step (5) data processing and survey report production include the following steps:
(1) statistical check data；
(2) only when blank sample inspection result is negative, control sample detection diatom type is identical as Known Species, diatom content is missed
When difference is less than given value 10-20%, sample inspection result is significant, just provides survey report；
(3) survey report is made, by Judicial Identification Document related request production survey report, in its check conclusion part,
The type and content of diatom in sample is stated, the matching degree of organs and tissues and water sample diatom examination result is also stated.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113702381A (en) * | 2021-09-06 | 2021-11-26 | 生强(天津)科技有限公司 | Diatom verifying attachment based on optical imaging |
| CN118777354A (en) * | 2024-07-16 | 2024-10-15 | 青海警官职业学院 | A method for detecting diatoms in the lungs of middle-aged and elderly deceased persons |
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