CN103776835A - Forensic medicine diatom detection method based on filter membrane - Google Patents
Forensic medicine diatom detection method based on filter membrane Download PDFInfo
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- CN103776835A CN103776835A CN201410039415.XA CN201410039415A CN103776835A CN 103776835 A CN103776835 A CN 103776835A CN 201410039415 A CN201410039415 A CN 201410039415A CN 103776835 A CN103776835 A CN 103776835A
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- 239000012528 membrane Substances 0.000 title claims abstract description 103
- 238000001514 detection method Methods 0.000 title abstract description 6
- 239000003814 drug Substances 0.000 title abstract 3
- 238000000034 method Methods 0.000 claims description 46
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 39
- 238000000967 suction filtration Methods 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 29
- RRAFCDWBNXTKKO-UHFFFAOYSA-N eugenol Chemical compound COC1=CC(CC=C)=CC=C1O RRAFCDWBNXTKKO-UHFFFAOYSA-N 0.000 claims description 26
- 239000003153 chemical reaction reagent Substances 0.000 claims description 25
- AHJKWBGXAGSAIK-UHFFFAOYSA-N acetic acid;2-methoxy-4-prop-2-enylphenol Chemical compound CC(O)=O.COC1=CC(CC=C)=CC=C1O AHJKWBGXAGSAIK-UHFFFAOYSA-N 0.000 claims description 15
- 238000007689 inspection Methods 0.000 claims description 15
- 238000003828 vacuum filtration Methods 0.000 claims description 15
- NPBVQXIMTZKSBA-UHFFFAOYSA-N Chavibetol Natural products COC1=CC=C(CC=C)C=C1O NPBVQXIMTZKSBA-UHFFFAOYSA-N 0.000 claims description 13
- 239000005770 Eugenol Substances 0.000 claims description 13
- UVMRYBDEERADNV-UHFFFAOYSA-N Pseudoeugenol Natural products COC1=CC(C(C)=C)=CC=C1O UVMRYBDEERADNV-UHFFFAOYSA-N 0.000 claims description 13
- 229960002217 eugenol Drugs 0.000 claims description 13
- 230000019771 cognition Effects 0.000 claims description 12
- 230000029087 digestion Effects 0.000 claims description 11
- 238000004451 qualitative analysis Methods 0.000 claims description 9
- 238000004445 quantitative analysis Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 210000000988 bone and bone Anatomy 0.000 claims description 7
- 230000007935 neutral effect Effects 0.000 claims description 7
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 7
- 239000012498 ultrapure water Substances 0.000 claims description 7
- 238000000879 optical micrograph Methods 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 5
- 239000004695 Polyether sulfone Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 229920006393 polyether sulfone Polymers 0.000 claims description 2
- 206010013647 Drowning Diseases 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 230000003287 optical effect Effects 0.000 abstract description 8
- 241000195493 Cryptophyta Species 0.000 abstract description 4
- 210000001519 tissue Anatomy 0.000 description 13
- 238000012545 processing Methods 0.000 description 11
- 210000004072 lung Anatomy 0.000 description 9
- 238000011084 recovery Methods 0.000 description 6
- 229920012266 Poly(ether sulfone) PES Polymers 0.000 description 5
- 239000006059 cover glass Substances 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 230000000007 visual effect Effects 0.000 description 5
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 2
- 238000011888 autopsy Methods 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- HQFCOGRKGVGYBB-UHFFFAOYSA-N ethanol;nitric acid Chemical compound CCO.O[N+]([O-])=O HQFCOGRKGVGYBB-UHFFFAOYSA-N 0.000 description 2
- 230000002440 hepatic effect Effects 0.000 description 2
- 210000005228 liver tissue Anatomy 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 210000005084 renal tissue Anatomy 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 241000206761 Bacillariophyta Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 208000010513 Stupor Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention relates to the field of algae detection, and particularly discloses a forensic medicine diatom detection method based on a filter membrane. After the filter membrane which is vacuum-filtered is transparently treated, the transparently treated filter membrane can be directly observed under an optical microscope. The detection method is simple to operate, high in detection efficiency, free from depending on complicated instruments, capable of detecting the diatom in a primary forensic medicine laboratory and wide in applicability.
Description
Technical field
The present invention relates to algae check field, be specifically related to a kind of diatom inspection method in medical jurisprudence based on filter membrane.
Background technology
Diatom is a kind of unicellular algae, and approximately there is kind more than 16000 in the whole world, and body is long generally between 1 μ m to 200 μ m, its the most obvious feature is that cell membrane is except indivedual kinds, all height silicidize, forms upper and lower two transparent shells, forms a siliceous cell wall with girdle band fit.The living environment of diatom is very extensive, and the trace of diatom almost can be seen in the waters of occurring in nature.
In water, corpse is type common in Medicolegal examination.Since the forties in 20th century, many legal medical expert scholars confirm on drowned person's corpse, and diatom can enter multiple internal organs by the circulation system.Frustule wall is difficult for by force destroyedly because of resistibility, and the high person of siliceous content, uses the concentrated sulphuric acid, red fuming nitric acid (RFNA) to boil even high temperature and burn also and be not destroyed.Therefore, diatom examination is a kind of reliable means that solves the cause of the death problem of corpse in water.
In medicolegal practice, 1～30g histoorgan is cleared up the rear digestion solution that conventionally can leave 10～50ml, and centrifugal is the common method of diatom in enrichment water sample or histoorgan digestion solution.In GA/T 813-2008 standards (" in human tissue organ diatom nitric acid broken machine method check "), pour postdigestive digestion solution into centrifuge tube film-making after repeatedly centrifugal; Finally adopt om observation method, with 10 times or 40 times of object lens inspection diatoms.Show according to the research such as Hu Sun Lin report, centrifugally once cause about 10% diatom loss, increase centrifugal speed and can not significantly improve the recovery of diatom, therefore cause in organs and tissues the diatom recovery low.
Because the method for vacuum filtration also can enrichment diatom, but filter membrane itself is opaque, cannot directly be placed in optical microphotograph Microscopic observation, and therefore Sun Lin etc. attempts using the method for vacuum filtration-automatic scanning Electronic Speculum to observe diatom recklessly.Although the resolution of electron microscope is high, it is clear to observe, but, in view of its complicated operation, cost is higher, maintenance cost is high feature, electron microscope can only be used in some big-and-middle-sized laboratories and scientific research institution, cannot be widely used in grass-roots method medical laboratory.
Summary of the invention
The technical problem to be solved in the present invention is the above-mentioned defect in order to overcome prior art, provide a kind of check highly sensitive, checkability is high, can significantly not increase again the diatom inspection method in medical jurisprudence based on filter membrane of equipment cost.
Object of the present invention is achieved by the following technical programs.
Based on a diatom inspection method in medical jurisprudence for filter membrane, comprise the steps:
S1. filter membrane enrichment: adopt vacuum filtration device that check sample is carried out to vacuum filtration; After sample suction filtration completes, add ultrapure water and continue suction filtration, filter membrane surface is approached neutral;
S2. film transparence: be placed on microslide dry filter membrane, drip acetic acid-eugenol reagent on filter membrane, covered;
S3. qualitative and quantitative analysis: the filter membrane after transparent is placed in to optical microphotograph Microscopic observation; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.
In prior art, adopt filter membrane concentration method that testing sample is carried out after suction filtration, because filter membrane itself is opaque, cannot directly be placed in optical microphotograph Microscopic observation, so must, by complicated facilities for observation as automatic scanning Electronic Speculum, the diatom on filter membrane be detected, the present invention is by suitable transparence processing mode, can make filter membrane transparent, can be placed directly in optical microphotograph Microscopic observation diatom.
The method of the invention is suitable for all types of detection samples, as the digestion solution of drown in water sample, tracheae liquid or histoorgan.
The dry processing that will first filter membrane be examined when filter membrane transparence is processed, as preferred embodiment, the dry method of described filter membrane is: the filter membrane after suction filtration is completed is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry.Filter membrane is carried out transparence processing by the acting as of acetic acid-eugenol reagent using in filter membrane transparence processing procedure.Inventor also tries to adopt acetic acid or eugenol reagent to carry out transparence processing to filter membrane in previous research work, but effect is all not so good as, with acetic acid-eugenol reagent, filter membrane is carried out to the effective of transparence processing.As preferred embodiment, described acetic acid-eugenol reagent is that the eugenol solution of the acetic acid solution of concentration 80～100% and concentration 60～100% is evenly mixed to the mixed solution obtaining in the ratio of 1:9～9:1.
In filter membrane enrichment process, can adopt simply connected or multi-joint vacuum filtration device to carry out vacuum filtration, simply connected vacuum filtration device, each is carried out suction filtration to a sample; Multi-joint vacuum filtration device can carry out suction filtration processing to multiple samples at every turn simultaneously.Preferably, when vacuum filtration, suction filtration vacuum tightness is: 100～800mmHg, and described filter membrane is the filter membrane of polyethersulfone material, filter membrane diameter is that 10～50mm, aperture are 0.1～1.0 μ m.
Compared with prior art, the present invention has following beneficial effect:
1, the recovery is high.Filter membrane enriching step of the present invention, has avoided the repeatedly loss of centrifugal treating diatom that process causes in traditional centrifugal enrichment method.The transparence reagent adopting can be by complete filter membrane transparence, and light can see through filter membrane in a large number to guarantee om observation.Observation by light microscope result shows, the filter membrane after transparence does not affect that it is quantitative.Therefore, compared with traditional centrifugal enrichment method, use filter membrane concentration method from water sample and organize enrichment diatom digestion solution, operating more easyly, the recovery is higher.
2, efficiency is high, cost performance is high, be easy to popularization.Filter membrane enriching step is easy and simple to handle, take a short time, and every kind of sample only needs enrichment once, and the repeatedly repeatedly centrifugal of traditional centrifugal enrichment method avoided in one of film-making, and repeatedly smear, film-making, both wasted time and energy, and makes again the recovery reduce.Film transparence step is simple, and the time is short, and the sample after transparent can directly be placed in optical microphotograph Microscopic observation, can guarantee the accuracy of assay, again without extra procuring equipment; Operating personnel only need simple training to operate.It is highly suitable for that funds are few, small scale but the laboratories that extensively distributes.
3, qualitative and quantitative analysis is accurate, labour intensity is low.Use the film-making of filter membrane concentration method to improve the diatom recovery, after film transparence, use observation by light microscope, diatom texture and details are clear, and do not affect that it is quantitative, have guaranteed the accuracy of qualitative and quantitative analysis.Film-making amount reduces, without too much training and operation, and can in conjunction with computer automation recognition method to captured picture check, classification and statistical treatment, these can significantly reduce analyst's labour intensity.
4, applicability is extensive.The present invention is not only applicable to diatom examination, more can extend to other check fields.Sample during as filter membrane enrichment can be selected plant counteract appearance, soil digested solusion, gas etc.; When transparence, can add suitable coloring agent, fixing agent, mark etc. to observe specific sample; Test rating capable of choosing multiple algae, small insects, parasite etc.Therefore, applicability of the present invention extensively, can interdisciplinaryly use.
Embodiment
Further describe the present invention below in conjunction with specific embodiment.Unless stated otherwise, reagent, the equipment that the present invention adopts is the conventional reagent of the art and equipment.
embodiment 1
A kind of diatom inspection method in medical jurisprudence based on filter membrane: be divided into three steps:
S1. filter membrane enrichment:
The sample of 50～100ml (the histoorgan digestion solution after dilution) is placed in to the interior suction filtration that starts of suction filtration cup of simply connected or multi-joint vacuum filtration device; Vacuum tightness 100～800mmHg, polyethersulfone (PES) filter membrane diameter 10～50mm, aperture is 0.1～1.0 μ m.Sample suction filtration adds the ultrapure water of 50～200ml to continue suction filtration while completing, filter membrane surface is approached neutral.
S2. film transparence:
Filter membrane after suction filtration completes is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry; Get the not microslide of use, center drips 1～5 acetic acid-eugenol reagent; The eugenol solution that acetic acid-eugenol reagent is concentration 80～100% acetic acid solutions and concentration 60～100% evenly mixes according to the ratio of 1:9 the mixed solution obtaining.Dried filter membrane is placed in to slide center, on filter membrane, again drips 1～3 acetic acid-eugenol reagent, add a cover cover glass.
S3. qualitative and quantitative analysis:
Slide after transparent is placed under optical microscope, with 20 times or 40 times of object lens observations; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.Under artificial cognition mode, as the miniature diatom in some visual field, because enlargement factor is limit, can not meet the requirement of evaluation, with 60 times of object lens even 100 times of oily mirrors observe its fine structure.
embodiment 2
A kind of diatom inspection method in medical jurisprudence based on filter membrane: be divided into three steps:
S1. filter membrane enrichment:
The sample of 50～100ml (the histoorgan digestion solution after dilution) is placed in to the interior suction filtration that starts of suction filtration cup of simply connected or multi-joint vacuum filtration device; Vacuum tightness 100～800mmHg, polyethersulfone (PES) filter membrane diameter 10～50mm, aperture is 0.1～1.0 μ m.Sample suction filtration adds the ultrapure water of 50～200ml to continue suction filtration while completing, filter membrane surface is approached neutral.
S2. film transparence:
Filter membrane after suction filtration completes is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry; Get the not microslide of use, center drips 1～5 acetic acid-eugenol reagent; The eugenol solution that acetic acid-eugenol reagent is concentration 80～100% acetic acid solutions and concentration 60～100% evenly mixes according to the ratio of 1:1 the mixed solution obtaining.Dried filter membrane is placed in to slide center, on filter membrane, again drips 1～3 acetic acid-eugenol reagent, add a cover cover glass.
S3. qualitative and quantitative analysis:
Slide after transparent is placed under optical microscope, with 20 times or 40 times of object lens observations; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.Under artificial cognition mode, as the miniature diatom in some visual field, because enlargement factor is limit, can not meet the requirement of evaluation, with 60 times of object lens even 100 times of oily mirrors observe its fine structure.
embodiment 3
A kind of diatom inspection method in medical jurisprudence based on filter membrane: be divided into three steps:
S1. filter membrane enrichment:
The sample of 50～100ml (the histoorgan digestion solution after dilution) is placed in to the interior suction filtration that starts of suction filtration cup of simply connected or multi-joint vacuum filtration device; Vacuum tightness 100～800mmHg, polyethersulfone (PES) filter membrane diameter 10～50mm, aperture is 0.1～1.0 μ m.Sample suction filtration adds the ultrapure water of 50～200ml to continue suction filtration while completing, filter membrane surface is approached neutral.
S2. film transparence:
Filter membrane after suction filtration completes is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry; Get the not microslide of use, center drips 1～5 acetic acid-eugenol reagent; The eugenol solution that acetic acid-eugenol reagent is concentration 80～100% acetic acid solutions and concentration 60～100% evenly mixes according to the ratio of 9:1 the mixed solution obtaining.Dried filter membrane is placed in to slide center, on filter membrane, again drips 1～3 acetic acid-eugenol reagent, add a cover cover glass.
S3. qualitative and quantitative analysis:
Slide after transparent is placed under optical microscope, with 20 times or 40 times of object lens observations; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.Under artificial cognition mode, as the miniature diatom in some visual field, because enlargement factor is limit, can not meet the requirement of evaluation, with 60 times of object lens even 100 times of oily mirrors observe its fine structure.
comparative example 1
A kind of diatom inspection method in medical jurisprudence based on filter membrane: be divided into three steps:
S1. filter membrane enrichment:
The sample of 50～100ml (the histoorgan digestion solution after dilution) is placed in to the interior suction filtration that starts of suction filtration cup of simply connected or multi-joint vacuum filtration device; Vacuum tightness 100～800mmHg, polyethersulfone (PES) filter membrane diameter 10～50mm, aperture is 0.1～1.0 μ m.Sample suction filtration adds the ultrapure water of 50～200ml to continue suction filtration while completing, filter membrane surface is approached neutral.
S2. film transparence:
Filter membrane after suction filtration completes is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry; Get the not microslide of use, center drips 1～5 acetic acid reagent; Acetic acid reagent is concentration 80～100% acetic acid solutions.Dried filter membrane is placed in to slide center, on filter membrane, again drips 1～3 acetic acid reagent, add a cover cover glass.
S3. qualitative and quantitative analysis:
Slide after transparent is placed under optical microscope, with 20 times or 40 times of object lens observations; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.Under artificial cognition mode, as the miniature diatom in some visual field, because enlargement factor is limit, can not meet the requirement of evaluation, with 60 times of object lens even 100 times of oily mirrors observe its fine structure.
Due to the poor effect that adopts 80～100% acetic acid solutions filter membrane to be carried out to transparence processing, the one, filter membrane is carried out to the chronic of transparence processing with 80～100% acetic acid solutions, use in addition the easy flavescence of 80～100% acetic acid solution filter membrane after treatment, be unfavorable for long-term preservation, so, be not suitable for adopting common optical microscope to observe.
comparative example 2
A kind of diatom inspection method in medical jurisprudence based on filter membrane: be divided into three steps:
S1. filter membrane enrichment:
The sample of 50～100ml (the histoorgan digestion solution after dilution) is placed in to the interior suction filtration that starts of suction filtration cup of simply connected or multi-joint vacuum filtration device; Vacuum tightness 100～800mmHg, polyethersulfone (PES) filter membrane diameter 10～50mm, aperture is 0.1～1.0 μ m.Sample suction filtration adds the ultrapure water of 50～200ml to continue suction filtration while completing, filter membrane surface is approached neutral.
S2. film transparence:
Filter membrane after suction filtration completes is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry; Get the not microslide of use, center drips 1～5 eugenol reagent; Eugenol reagent is the eugenol solution of concentration 60～100%.Dried filter membrane is placed in to slide center, on filter membrane, again drips 1～3 eugenol reagent, add a cover cover glass.
S3. qualitative and quantitative analysis:
Slide after transparent is placed under optical microscope, with 20 times or 40 times of object lens observations; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.Under artificial cognition mode, as the miniature diatom in some visual field, because enlargement factor is limit, can not meet the requirement of evaluation, with 60 times of object lens even 100 times of oily mirrors observe its fine structure.
Due to the poor effect that adopts 60～100% eugenol solution filter membrane to be carried out to transparence processing, the one, filter membrane is carried out to the chronic of transparence processing with 60～100% eugenol solution, use in addition the easy flavescence of filter membrane after 60～100% eugenol solution-treated, be unfavorable for long-term preservation, so, be not suitable for adopting common optical microscope to observe.
application examples
Described in use embodiment 1, method detects analysis to died:
Case 1:
On September 21st, 2013, find a tool female corpse on the limit, pond that certain primary school's lateral gate is other, approximately 20 years old age, with it without any certificate article.After corpse table look-up, be difficult to determine the cause of the death and case character.
When diatom examination, adopt nitric acid Ethanol Method to clear up, centrifugal enrichment method is in conjunction with observation by light microscope, result from lung, detect with pond in the diatom of same type, content is 1086/10g lung tissue, in liver and renal tissue, does not detect diatom.Then will clear up a kind of filter membrane enrichment of rear use, transparence, om observation technical checking diatom with a tissue samples, from the dead's lung, liver, kidney, all detect with pond water sample in the diatom of same type, content is respectively 1275/10g lung tissue, 61/10g hepatic tissue, 8/10g nephridial tissue, therefore identifies that this women's cause of the death is for being drowned.In conjunction with other investigation result, finally confirm that the dead is unexpected overboard death.
Case 2:
On March 6th, 2013, on the other river levee of certain middle school's basketball court, find the unknown female's corpse of a tool, corpse height corruption, for confirming that the dead is drowned or is after death entered water by throwing corpse in being thrown into water before death, after autopsy, get lung, liver and nephridial tissue, adopt nitric acid Ethanol Method to clear up, centrifugal enrichment method is in conjunction with observation by light microscope, result from lung, detect with river in the diatom of same type, content is 1762/10g lung tissue, in liver and renal tissue, does not detect diatom.Then will clear up a kind of filter membrane enrichment of rear use, transparence, om observation technical checking diatom with a tissue samples, from the dead's lung, liver, all detect with river in the diatom of same type, content is respectively 1265/10g lung tissue, 11/10g hepatic tissue, nephridial tissue does not detect diatom, according to this assay, in conjunction with autopsy finding, identify that this women's cause of the death is for being drowned.After cracking of cases, confirm: the dead walks to certain doorway, middle school evening on March 1st, 2013, Lee who hides in the shadow finds one people walking and carries with oneself a knapsack on the arm.Lee is leapt up out suddenly and is robbed Qi Bao, and in the process battling with the dead, goner " chokes " (reality is only the of short duration stupor of suffocating) to death, and near the then korneforos of goner pulling into, with the dead's bag escape.
Claims (5)
1. the diatom inspection method in medical jurisprudence based on filter membrane, is characterized in that, comprises the steps:
S1. filter membrane enrichment: adopt vacuum filtration device that check sample is carried out to vacuum filtration; After sample suction filtration completes, add ultrapure water and continue suction filtration, filter membrane surface is approached neutral;
S2. film transparence: be placed on microslide dry filter membrane, drip acetic acid-eugenol reagent on filter membrane, covered;
S3. qualitative and quantitative analysis: the filter membrane after transparent is placed in to optical microphotograph Microscopic observation; Adopt artificial cognition mode or computer automation recognition method to the diatom on filter membrane or captured picture check, classification and statistical treatment.
2. the diatom inspection method in medical jurisprudence based on filter membrane according to claim 1, is characterized in that, check sample is the digestion solution of liquid or histoorgan of drowning in water sample, tracheae described in S1.
3. the diatom inspection method in medical jurisprudence based on filter membrane according to claim 1, is characterized in that, the dry method of filter membrane is described in S2: the filter membrane after suction filtration is completed is placed on the electric furnace of 50～80 ℃ or baking oven heats 1～10min and makes its bone dry.
4. the diatom inspection method in medical jurisprudence based on filter membrane according to claim 1, it is characterized in that, the acetic acid solution that acetic acid-eugenol reagent is concentration 80～100% described in S2 and the eugenol solution of concentration 60～100% evenly mix in the ratio of 1:9～9:1 the mixed solution obtaining.
5. the diatom inspection method in medical jurisprudence based on filter membrane according to claim 1, is characterized in that, suction filtration vacuum tightness is described in S1: 100～800mmHg, and described filter membrane is the filter membrane of polyethersulfone material, filter membrane diameter is that 10～50mm, aperture are 0.1～1.0 μ m.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105784705A (en) * | 2016-02-21 | 2016-07-20 | 山东政法学院 | Forensic medicine diatom test method |
| CN110033445A (en) * | 2019-04-10 | 2019-07-19 | 司法鉴定科学研究院 | Medicolegal examination automatic identification system and recognition methods based on deep learning |
| CN113252413A (en) * | 2021-04-16 | 2021-08-13 | 成都新基因格生物科技有限公司 | Diatom inspection method |
| CN115406739A (en) * | 2022-07-22 | 2022-11-29 | 华南农业大学 | Preparation method of permanent mounting sheet for phytoplankton quantitative analysis sample |
| CN116990297A (en) * | 2023-09-27 | 2023-11-03 | 佳木斯大学 | Forensic science diatom check-out set based on filter membrane |
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Cited By (8)
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|---|---|---|---|---|
| CN105784705A (en) * | 2016-02-21 | 2016-07-20 | 山东政法学院 | Forensic medicine diatom test method |
| CN105784705B (en) * | 2016-02-21 | 2018-05-18 | 山东政法学院 | A kind of diatom inspection method in medical jurisprudence |
| CN110033445A (en) * | 2019-04-10 | 2019-07-19 | 司法鉴定科学研究院 | Medicolegal examination automatic identification system and recognition methods based on deep learning |
| CN113252413A (en) * | 2021-04-16 | 2021-08-13 | 成都新基因格生物科技有限公司 | Diatom inspection method |
| CN113252413B (en) * | 2021-04-16 | 2023-04-11 | 成都新基因格生物科技有限公司 | Diatom inspection method |
| CN115406739A (en) * | 2022-07-22 | 2022-11-29 | 华南农业大学 | Preparation method of permanent mounting sheet for phytoplankton quantitative analysis sample |
| CN116990297A (en) * | 2023-09-27 | 2023-11-03 | 佳木斯大学 | Forensic science diatom check-out set based on filter membrane |
| CN116990297B (en) * | 2023-09-27 | 2024-01-02 | 佳木斯大学 | Forensic science diatom check-out set based on filter membrane |
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