CN103536296B - Method of detecting human sweat secretion - Google Patents
Method of detecting human sweat secretion Download PDFInfo
- Publication number
- CN103536296B CN103536296B CN201310493627.0A CN201310493627A CN103536296B CN 103536296 B CN103536296 B CN 103536296B CN 201310493627 A CN201310493627 A CN 201310493627A CN 103536296 B CN103536296 B CN 103536296B
- Authority
- CN
- China
- Prior art keywords
- fingerprint
- finger mark
- antibody
- target
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)
Abstract
The invention discloses a method of detecting human sweat secretion. The method includes: transferring a fingerprint to a substrate to obtain a fingerprint sample; adding nano-gold labeled target secretion antibody solution to be detected to the fingerprint of the fingerprint sample, incubating, and then rinsing with rinsing solution; drying the fingerprint sample, adding slivering solution to the fingerprint, developing in the shade, rinsing and drying the fingerprint sample, and judging; if the fingerprint appears, determining the fingerprint contains the target secretion to be detected; if the fingerprint not appearing, determining the fingerprint contains no target secretion to be detected. The method of detecting the human sweat secretion is capable of detecting the target secretion simply, quickly and specifically, and also capable of identifying an object containing the target secretion.
Description
Technical field
The invention belongs to bioanalysis and fingerprinting detection technique field, particularly relate to a kind of method of human body sweat secretion thing.
Background technology
Fingerprint is the lines that human finger's end refers to abdomen is formed by concavo-convex skin, is formed and shape in skin development process, has special physiological structure and feature architecture.In the evolutionary process of the mankind, the physiological use of fingerprint makes hands increase frictional force when contact subject, thus more easily have an effect and firmly grasp object.People were again according to its specificity and invariance afterwards, were applied to the person and identified and forensic identification, and extend to the field such as safety verification, personal authentication in daily life.
Can finger mark according to be directly divided into obvious finger mark and potential finger mark as seen by naked eyes, and obvious finger mark is generally stained with the article transfer printings such as paint, blood, ink and is formed by hands; Potential finger mark is then that the fingerprint lines that transfer is formed, visually not easily finding, is modal fingerprint in spot through health nature secretions as perspiration.One piece of common antiperspirant finger mark is about 0.11mg, and wherein 99% is water, and have half to be the inorganic matters such as sodium chloride in remaining 1%, second half is then the Organic substances such as oils and fats, lysozyme, aminoacid.These micro-content organisms allow scientist manifest at the optics of routine and outside physisorphtion, to develop the method that a lot of chemistry manifests fingerprint just.The detection that Recent study person utilizes mass spectrum imaging, infrared/new technique such as Raman image and immune labeled imaging carries out fingerprint manifestation and fingerprint composition, thus excavate out more how valuable biology and medical information.Such as, whether the whether contacted explosive of terrorist, a people have the custom of smoking, and scientist can obtain relevant information by fingerprint detection.
Saunders(74th Annual Enducational Conference in 1989, Penacola, USA, 1989) propose to manifest potential finger mark by many metalidings.Its principle is soaked by there being the sample of finger mark to put into citric acid gold, after a period of time, nano Au particle wherein will be adsorbed onto on the ridge of finger mark, then with silver-colored dye liquor development sample, because the reduction of nanometer gold to silver ion has the ability of catalysis, so silver-colored simple substance can be separated out faster on ridge, the ridge blackening overstriking of finger mark is displayed.Selectivity when nanometer gold adsorbs finger mark in the method directly impact can manifest effect, and the factor affecting result is a lot, as pH value of solution.A lot of scientist was had again to improve (Chinese patent being CN 101703404A as publication number is total to disclosed " manifesting multiple object fingerprint surface and the method for reservation DNA information ") the method afterwards, what utilize is all hydrophilic and hydrophobic and the Electrostatic Absorption of nanometer gold and finger mark, but still non-specific adsorption is serious, affect fingerprint imaging effect, and complex operation.
Summary of the invention
The invention provides a kind of method of human body sweat secretion thing, detection target secretions that can be simple, quick, special, also can carry out the person to the object containing target secretions simultaneously and identify.
A method for human body sweat secretion thing, comprising:
(1) Sweat latent fingerprint is transferred in substrate, obtain finger mark sample;
(2) the target secretions antibody-solutions to be detected adding nano gold mark on the Sweat latent fingerprint of described finger mark sample is hatched, and has hatched rear wash solution and has rinsed;
(3) described finger mark sample is dry, then on Sweat latent fingerprint, add silver-colored dye liquor, after lucifuge colour developing, by the rinsing, drying of finger mark sample, judge:
If display fingerprint, then contain target secretions to be detected in Sweat latent fingerprint; Otherwise, then target secretions to be detected is not contained in Sweat latent fingerprint.
When finger skin touches object, the secretions on skin will be left a trace on its surface, forms the mirror image figure of fingerprint.The ridge line region that finger mark is made up of sweat mixed with dirt and grease and blank line region, valley form.The ridge line of fingerprint, except most of moisture, also contains the human secretions such as organic aliphatic acid, aminoacid, lysozyme and epidermal growth factor.
2006, the forensic specialist David Russell of east England university of Britain once proposed, can secrete cotinine (a kind of metabolite of nicotine) in the perspiration of smoker, appreciation personnel can by detecting the cotinine in fingerprint whereby, judges fingerprint owner's whether smoking.2007, Russell seminar successfully achieved this imagination, and it have also demonstrated in the perspiration of drug addict and can there is relevant drugs metabolite afterwards.Therefore, if people has smoking, custom etc. of taking drugs, in its perspiration, just can secrete the characteristic metabolic thing corresponding to cigarette, drugs.
Therefore, the application utilizes immunoreactive specific recognition mechanism to realize metabolite and detects the combination identified with the person.If containing target metabolite to be detected in Sweat latent fingerprint, then carry the nanogold particle of target secretions antibody to be detected and be orientablely attached on fingerprint ridge line, profile and the structure of fingerprint just can be distinguished through the colour developing of silver-colored simple substance deposition, show visual fingerprint lines, also can reach the object of person identification further according to the fingerprint manifested simultaneously, if do not manifest fingerprint after the method process of the application, then do not contain target metabolite to be detected in Sweat latent fingerprint.
In step (3), after the rinsing, drying of finger mark sample, if fingerprint manifestation, then read the fingerprint characteristic of finger mark sample further by scan-type electrochemical microscope.Scan-type electrochemical microscope (SECM) on the ridge line containing silver-colored simple substance and the hardly line place, valley of argentiferous scans the size of current obtained and has difference, geographic location feature can be changed into current signal, amplify details in fingerprint after being further converted to image.SECM can be accurate to nano-grade size, can amplify details in fingerprint to greatest extent.
As the carrier of finger mark, the application is not strict with substrate, and described substrate is glass plate, plastic plate, corrosion resistant plate, tinfoil paper, hard paper, adhesive tape, alloy, microscope slide, conductive copper subsides, Mobile phone film, candy wrapper etc.
Described target secretions to be detected is epidermal growth factor, lysozyme or cotinine.
The concentration of the target secretions antibody-solutions to be detected of described nano gold mark is 0.05 ~ 0.2mg/mL, is preferably 0.1 ~ 0.2mg/mL.The excessive concentration of antibody-solutions not only wastes reagent, also can cause non-specific binding, or colour developing background weight, and concentration is too low, then the reaction of antigen and antibody can be caused incomplete, silver dye weak effect, thus affects fingerprint imaging effect.Wherein, the particle diameter of nanometer gold is for being generally 15nm.
The solvent of the target secretions antibody-solutions to be detected of described nano gold mark is be 0.01M, pH containing the concentration of 2%BSA, the PBS of 0.1% tween 20, PBS is 7.4.The adsorption of antibody to substrate can be reduced after adding appropriate BSA and tween 20 in PBS, make fingerprint image more clear.
Carry out for reacting fully, described in time of hatching be 10-45min, be preferably 15min.
Described wash solution is the PBS containing 0.1% tween 20.
Described silver-colored dye liquor is Sigma-aldrich silver enhancement solution, and purchase company is Sigma-aldrich, comprises A liquid and B liquid, and wherein A liquid article No. is S5020, B liquid article No. is S5145, by A liquid and the 1:1 mixing by volume of B liquid during use.
The time of described lucifuge colour developing is 15 ~ 20min, is preferably 15min.The long too short blur-free imaging being all unfavorable for fingerprint of developing time, if the time, too short reaction was insufficient, the time, oversize then background was excessively dark.
Compared with prior art, beneficial effect of the present invention is:
(1) method of the present invention deposits based on immunity and many metals, specificity is high, and by the fingerprint manifested, person identification is carried out to the object containing target secretions, and fingerprint manifestation is clear, by being observed visually the firsts and seconds structure of detail characteristics of fingerprints after fingerprint manifestation, when target secretion content is high, tertiary structure sweat pore can be observed.
(2) scanning electrochemical microscope (SECM)) (SECM) was invented in 1989, and was able to commercialization.What SECM measured is be oxidized or reduce the electrochemical source of current provided by chemical substance, and sample is not restricted to conductor, also can be insulator or quasiconductor.SECM, except providing except the pattern of sample surfaces, can also provide abundant chemical information.Method of the present invention can overcome the interference of dark-background after utilizing SECM to manifest, and to the electric conductivity no requirement (NR) of substrate, and clearly can manifest three grades of sweat pore structures, is the important evidence of incomplete finger mark identification.
(3) the present invention is in fingerprint manifestation compared with the traditional method such as brush dust, and required medicine is poisoned little, is conducive to the safety protecting operator and the pollution reduced environment.
(4) in sweat secretion thing context of detection, the method and mass spectrum, infrared etc. compared with, there is the visual advantage such as seen of, result auxiliary, easy and simple to handle without the need to large-scale instrument.
(5) compared with fluorescence method, method of the present invention does not need external excitation light source, and therefore there is not bias light interference, fingerprint image is more significantly clear.
(6) method of the present invention can obtain the identity information of testee while to human metabolite test, therefore can promote the use of in the picket of illegal drug drugs, criminal investigation and routine safety as poison is driven, in the supervision of drunk driving.
Accompanying drawing explanation
Fig. 1 is the operational flowchart that human body sweat secretion quality testing is surveyed.
Fig. 2 is the structural representation of SECM imaging system;
Fig. 3 is the structural representation in example reaction pond in the SECM imaging system shown in Fig. 2;
Wherein, 1, sample reaction tank; 2, bipotentiostat controller; 3,3 D locating device; 4, base; 5, cell body; 6, groove; 7, O RunddichtringO; 8, bolt; 9, electrode hole; 10, working electrode; 11, to electrode; 12, reference electrode; 13, computer.
Fig. 4 a is that the goat anti-human igg antibody's (PBS dilution) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 4 b is that the goat anti-human igg antibody's (PBS-T-BSA dilution) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 5 a is that the goat anti-human igg antibody's (concentration is 0.02mg/mL) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 5 b is that the goat anti-human igg antibody's (concentration is 0.20mg/mL) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 5 c is that the goat anti-human igg antibody's (concentration is 0.40mg/mL) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 6 a is that the goat anti-human igg antibody's (developing time is 10min) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 6 b is that the goat anti-human igg antibody's (developing time is 15min) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 6 c is that the goat anti-human igg antibody's (developing time is 20min) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 7 a is that the goat anti-human igg antibody's (incubation time is 5min) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 7 b is that the goat anti-human igg antibody's (incubation time is 15min) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 7 c is that the goat anti-human igg antibody's (incubation time is 60min) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 8 a is that the goat anti-human igg antibody's (incubation temperature is room temperature 20 ± 2 DEG C) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 8 b is that the goat anti-human igg antibody's (incubation temperature is 37 DEG C) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 9 a is that the goat anti-human igg antibody's (wash solution is PBS) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Fig. 9 b is that the goat anti-human igg antibody's (wash solution is PBS-T) with nano gold mark detects the immunoglobulin G while simulation finger mark silver dye colour developing figure obtained.
Figure 10 is the latent fingerprint silver dye colour developing figure obtained with the cotinine antibody test of nano gold mark.
Figure 11 is the latent fingerprint silver dye colour developing figure obtained with the lysozyme antibody test of nano gold mark.
Figure 12 is that epidermal growth factor (EGF) conduct detects object, through the post-depositional fingerprint image of the many metals of immunity.
Figure 13 is that immunoglobulin G while (hIgG) simulates finger mark through the post-depositional image of the many metals of immunity and with the image after SECM partial enlargement.
Figure 14 is that immunoglobulin G while (hIgG) simulates finger mark on different substrates through the post-depositional image of the many metals of immunity.
Detailed description of the invention
The present invention is explained further below in conjunction with detailed description of the invention.
SECM imaging system can adopt conventional structure, and specifically purchased from Shanghai Chen Hua instrument company, model is CHI920C.
As shown in Figures 2 and 3, SECM imaging system comprises sample reaction tank 1, bipotentiostat controller 2, high-resolution 3 D locating device 3 and computer 13.
Bipotentiostat controller 2 comprises working electrode 10, to electrode 11 and reference electrode 12, computer 13 manipulates whole process, and wherein, working electrode is the ultramicroelectrode (Pt) of diameter 25 μm, is platinum filament to electrode, and reference electrode is Ag electrode.
3 D locating device 3 is made up of motor and piezoquartz, can carry out three-dimensional localization to ultramicroelectrode, allows the range ability of 50 millimeters and reaches the spatial discrimination of nanometer.
Sample reaction tank 1 comprises base 4 and the cell body 5 for holding reactant liquor, all can adopt polytetrafluoroethylmaterial material or other materials that can not be corroded by reactant liquor.Base 4 has the groove 6 for sample.
Cell body 5 is fixed on base 4 by bolt 8, the both sides of cell body 5 are respectively provided with an electrode hole 9, during work, reference electrode 12 is placed therein in an electrode hole, be positioned in another electrode hole to electrode 11, cell body 5 hollow, the hollow part of cell body 5 is just to groove 6, and this hollow part is provided with O RunddichtringO 7.
Post-depositional for many for immunity metals finger mark sample is fixed in the groove 6 of sample reaction tank base of SECM imaging system, in cell body 5, add reactant liquor (reactant liquor is the Alkitrate (0.1M) containing 1mM potassium hexachloroiridate), reactant liquor liquid level wants can not have reference electrode and to electrode; Be positioned over by reaction tank on SECM platform, under " probe convergence pattern ", the current potential of applying ± 0.8V is surperficial near fingerprint by probe; Determine the region needing to carry out scanning, select " probe scanning pattern " respectively along X and Y-direction scanning, regulate platform to make it be tending towards level according to the variation tendency of single line electric current as far as possible." scan-type electrochemical microscope pattern " is finally selected to gather details in fingerprint image.
Below in conjunction with drawings and Examples, the present invention is described in more detail.
The present invention can manifest finger mark based on the many metal depositions of immunity and SECM and detect target secretions, the sample carrying antiperspirant finger mark is hatched through the antibody-solutions of decorated by nano-gold, after washing argon dries up, add the development of silver-colored dye liquor, rinse with water after sucking silver-colored dye liquor with filter paper and dry up.Then be fixed in reaction tank, add reactant liquor, scanning collection fingerprint image after applying current potential.
In the present invention, the probe of SECM first moves to fingerprint surface, then in the enterprising line scanning of the plane of X-Y.Because probe is very close to fingerprint, when moving to fingerprint diverse location, its electric current because the silver-colored simple substance on fingerprint ridge increases the facilitation of reacting in solution, can obtain the image that generation current is relevant to position.The point of each electric current different colours is represented, just can reduce details in fingerprint.
The cotinine antibody of nano gold mark is synthesized by Hangzhou Long Ji Bioisystech Co., Ltd; The goat-anti hEGF antibody of goat anti-human igg antibody's nano gold mark of nano gold mark and the lysozyme antibody of nano gold mark are synthesized by Shanghai Sheng Gong Biological Co., Ltd., wherein, nanometer gold is the golden nanometer particle of the citric acid modification of particle diameter 15nm, the article No. of goat anti-human igg antibody is DAC1011(Sheng Gong biological engineering company limited), the article No. of lysozyme antibody is DAC1011(Sheng Gong biological engineering company limited), the article No. of goat-anti hEGF antibody is DA1709.
Silver dye liquor: purchase company is Sigma-aldrich, comprises A liquid and B liquid, and wherein, A liquid article No. is S5020, B liquid article No. is S5145.
Embodiment 1
1, antibody solvent is on the impact of fingerprint imaging
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, in reaction tank, adding goat anti-human igg antibody's solution of 100 μ L 0.1mg/mL nano gold marks, (solvent is for containing 2%BSA, the 0.01M PBS of 0.1% tween 20, pH are 7.4; With pH be 7.4 PBS be contrast), room temperature hatches 15min;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.
As shown in figures 4 a and 4b, adopt PBS to dilute the goat anti-human igg antibody of nano gold mark, antibody is serious (Fig. 4 a) to substrate absorption, fingerprint imaging is unintelligible, with the addition of tween and BSA in PBS after, considerably reduce its absorption to substrate, fingerprint texture clear (Fig. 4 b).
2, goat anti-human igg antibody's solution of the nano gold mark of variable concentrations is on the impact of fingerprint imaging
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, in reaction tank, adding goat anti-human igg antibody's solution of 100 μ L nano gold marks, (solvent is for containing 2%BSA, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch 15min in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.
The concentration that Fig. 5 a-5c shows goat anti-human igg antibody's solution of nano gold mark is respectively 0.02,0.20,0.40mg/mL time the fingerprint image that gathers, the concentration of goat anti-human igg antibody's solution of nano gold mark is too low (0.02mg/mL), and silver dye color developing effect not obviously (Fig. 5 a); Concentration too high (0.40mg/mL) can make background color deepen (Fig. 5 c); The color developing effect of 0.20mg/mL is best (Fig. 5 b).
3, silver dye developing time is on the impact of fingerprint imaging
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, goat anti-human igg antibody's solution of 100 μ L 0.1mg/mL nano gold marks is added (containing 2%BSA in reaction tank, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch 15min in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.
Fig. 6 a-6c show lucifuge developing time be respectively 10,15,25min time the fingerprint image that gathers, the silver-colored dye time too short (10min), color is more shallow, and colour developing is dark not (Fig. 6 a); The silver dye time oversize (25min), background color can be made to deepen (Fig. 6 c); Colour developing 15min comparatively suitable (Fig. 6 b).
4, incubation time is on the impact of fingerprint imaging
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, goat anti-human igg antibody's solution of 100 μ L 0.1mg/mL nano gold marks is added (containing 2%BSA in reaction tank, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.
Fig. 7 a-7c show incubation time be respectively 5,15,60min time the fingerprint image that gathers, the silver-colored dye time too short (5min), color is more shallow, and colour developing is dark not (Fig. 7 a); Incubation time oversize (60min), can make non-specific adsorption increase and lose time (Fig. 7 c); Hatch 15min comparatively suitable (Fig. 7 b).
5, incubation temperature is on the impact of fingerprint imaging
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, goat anti-human igg antibody's solution of 100 μ L 0.1mg/mL nano gold marks is added (containing 2%BSA in reaction tank, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch 15min;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.
Fig. 8 a-8b shows the fingerprint image gathered when incubation temperature is respectively room temperature (20 ± 2 DEG C) and 37 DEG C, under finding normal temperature condition, (time Fig. 8 a) He 37 DEG C (Fig. 8 b), silver dye effect does not have notable difference, therefore hatches from simple experiment considers to select room temperature.
6, detergent solution is on the impact of fingerprint imaging
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, goat anti-human igg antibody's solution of 100 μ L 0.1mg/mL nano gold marks is added (containing 2%BSA in reaction tank, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch 15min in room temperature;
Hatched rear wash solution to rinse to wash away unconjugated antibody, dried up on surface with argon, then added 200 μ L silver dye liquor (A:B=1:1), in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.
Fig. 9 a-9b shows wash solution and is respectively PBS(0.01M, pH7.4) and PBS-T(0.01M containing the PBS of 0.1% tween 20, pH7.4) fingerprint image gathered time, the image using the colour developing image (Fig. 9 b) obtained containing the wash solution of tween-20 to obtain than the wash solution not containing tween-20 (Fig. 9 a) in secondary and tertiary structure more clear.
Embodiment 2 manifests finger mark based on the many metal depositions of immunity and detects the cotinine in fingerprint
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of Sweat latent fingerprint sample: smoker will hand-wash only with fancy soap, after hot blast drying, puts on PE glove and seals antiperspirant 5min, then in substrate, press 1min, obtain Sweat latent fingerprint.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, in reaction tank, adding the cotinine antibody-solutions of 100 μ L 0.1mg/mL nano gold marks, (solvent is for containing 2%BSA, the 0.01M PBS of 0.1% tween 20, pH is 7.4), cultivate 15min in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.Image is as Figure 10, and visible fingerprint image texture is high-visible.
Embodiment 3 manifests finger mark based on the many metal depositions of immunity and detects the lysozyme in fingerprint
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.(2) preparation of Sweat latent fingerprint sample: tester will hand-wash only with fancy soap, after hot blast drying, puts on PE glove and seals antiperspirant 5min, then in substrate, press 1min, obtain Sweat latent fingerprint.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, in reaction tank, adding the lysozyme antibody-solutions of 100 μ L 0.1mg/mL nano gold marks, (solvent is for containing 2%BSA, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch 15min in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing;
Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera, and image is as Figure 11, and visible fingerprint image texture is high-visible.
Embodiment 4 manifests finger mark based on the many metal depositions of immunity and detects the epidermal growth factor in fingerprint
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of Sweat latent fingerprint sample: smoker will hand-wash only with fancy soap, after hot blast drying, puts on PE glove and seals antiperspirant 5min, then in substrate, press 1min, obtain Sweat latent fingerprint.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, in reaction tank, adding epidermal growth factor (EGF) antibody-solutions of 100 μ L 0.1mg/mL nano gold marks, (solvent is for containing 2%BSA, the 0.01M PBS of 0.1% tween 20, pH is 7.4), cultivate 15min in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or to take pictures collection image with digital camera.Image is as Figure 12, and visible fingerprint image texture is high-visible.
Embodiment 5 manifests finger mark based on the many metal depositions of immunity and SECM and detects immunoglobulin G while simulation finger mark
(1) base treatment
Substrate is that professional finger mark extracts adhesive tape, and it does not have the one side double faced adhesive tape of stickiness to be fixed on microscope slide, it can be used as supporter.
(2) preparation of immunoglobulin G while simulation finger mark sample:
First experimenter will hand-wash only with soap, dry up after deionized water rinsing with hair-dryer, and finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on adhesive tape after dry, obtain the albumen finger mark of simulating.
(3) immune many metals are deposited as picture: around finger mark, draw a circle with SABC pen, form a reaction tank, spread to avoid antibody-solutions, goat anti-human igg antibody's solution of 100 μ L 0.1mg/mL nano gold marks is added (containing 2%BSA in reaction tank, the 0.01M PBS of 0.1% tween 20, pH is 7.4), hatch 15min in room temperature;
Hatch rear wash solution (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) rinse to wash away unconjugated antibody, with argon, surface is dried up, add 200 μ L silver dye liquor (A:B=1:1) again, in darkroom, lucifuge colour developing 15min, dries up with argon after deionized water rinsing; Sample after colour developing can directly with the naked eye be observed, or takes pictures with digital camera and gather to obtain image shown in (a) in Figure 13.
(4) SECM amplifies finger mark details
Post-depositional for many for immunity metals finger mark sample is fixed in the sample reaction tank of SECM imaging system, reactant liquor (reactant liquor is the Alkitrate (0.1M) containing potassium hexachloroiridate (1mM)) is added in cell body, to be positioned on SECM platform and to regulate and make its level, the current potential applying 0.8V is surperficial near fingerprint by probe, select the region needing to carry out scanning, scanning collection details in fingerprint image, as shown in (b) in Figure 13, can obtain current value everywhere as shown in (c) in Figure 13 simultaneously.
Embodiment 6 manifests based on the many metal depositions of immunity and detects the immunoglobulin G while simulation finger mark in different base
(1) finger mark sample preparation
(tinfoil paper on different objects, alloy, microscope slide, conductive copper is pasted, Mobile phone film, candy wrapper) first experimenter will hand-wash only with soap, dry up with hair-dryer after deionized water rinsing, finger is coated with the immunoglobulin G while solution 25 μ L spreading 1mg/mL, gently by 2min on each object after dry, obtains the albumen finger mark of simulating.
(2) immune many metals are deposited as picture
Around the finger mark of finger mark sample, a circle is drawn with SABC pen, form a reaction tank, spread to avoid antibody-solutions, in reaction tank, add 100 μ L contain the goat anti-human immunoglobulin G antibody-solutions of 0.2mg/mL nano gold mark (solvent is 0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) Tween-20,2%(w/v) phosphate buffered solution of BSA), room temperature hatches 15min;
Hatch rear detergent (0.01M, pH 7.4, containing 0.15M NaCl, 0.1%(v/v) phosphate buffered solution of Tween-20) wash finger mark sample 3 times after dry up, add 200 μ L silver dye liquor (A:B=1: 1) colour developing 15min, filter paper blots, image shown in Figure 14 is obtained with collected by camera, wherein (a): tinfoil paper, (b): alloy, (c): microscope slide, (d): conductive copper is pasted, (e): Mobile phone film, (f): candy wrapper, visible in figure, although object is different, adopt the method for the application all can obtain fingerprint image clearly.
Claims (5)
1. a method for human body sweat secretion thing, is characterized in that, comprising:
(1) Sweat latent fingerprint is transferred in substrate, obtain finger mark sample;
(2) the target secretions antibody-solutions to be detected adding nano gold mark on the Sweat latent fingerprint of described finger mark sample carries out hatching 10-45min, has hatched rear wash solution and has rinsed;
(3) described finger mark sample is dry, then on Sweat latent fingerprint, add silver-colored dye liquor, after lucifuge colour developing, by the rinsing, drying of finger mark sample, judge:
If display fingerprint, then contain target secretions to be detected in Sweat latent fingerprint; Otherwise, then target secretions to be detected is not contained in Sweat latent fingerprint;
Described target secretions to be detected is epidermal growth factor, lysozyme;
The concentration of the target secretions antibody-solutions to be detected of described nano gold mark is 0.05 ~ 0.2mg/mL;
The solvent of the target secretions antibody-solutions to be detected of described nano gold mark is for containing 2%BSA, the PBS of 0.1% tween 20.
2. the method for claim 1, is characterized in that, in step (3), if display fingerprint, is read the fingerprint characteristic of finger mark sample by scan-type electrochemical microscope.
3. the method for claim 1, is characterized in that, described substrate is glass plate, plastic plate, corrosion resistant plate, tinfoil paper, hard paper or adhesive tape.
4. the method for claim 1, is characterized in that, described wash solution is the PBS containing 0.1% tween 20.
5. the method for claim 1, is characterized in that, the time of described lucifuge colour developing is 15 ~ 20min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310493627.0A CN103536296B (en) | 2013-10-18 | 2013-10-18 | Method of detecting human sweat secretion |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310493627.0A CN103536296B (en) | 2013-10-18 | 2013-10-18 | Method of detecting human sweat secretion |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103536296A CN103536296A (en) | 2014-01-29 |
| CN103536296B true CN103536296B (en) | 2015-06-17 |
Family
ID=49960442
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310493627.0A Active CN103536296B (en) | 2013-10-18 | 2013-10-18 | Method of detecting human sweat secretion |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103536296B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103837592B (en) * | 2014-03-18 | 2016-08-31 | 中国计量科学研究院 | A kind of method of iron content in isotopic dilution LA-ICP-MS in-site detecting biological tissue samples |
| CN104076025B (en) * | 2014-06-18 | 2017-04-05 | 深圳职业技术学院 | A kind of antibacterial peptide electrochemical luminous sensor and preparation method thereof and detection method |
| GB2528654B (en) * | 2014-07-24 | 2017-01-04 | Intelligent Fingerprinting Ltd | Drug dispenser assembly |
| GB2528657B (en) * | 2014-07-24 | 2019-03-13 | Intelligent Fingerprinting Ltd | Sample analysing device |
| CN105445251A (en) * | 2014-08-14 | 2016-03-30 | 杭州坚峰科技有限公司 | Spectral probe and preparation method and use thereof |
| CN104964960B (en) * | 2015-06-08 | 2017-07-07 | 江南大学 | A kind of method of the detection VEGF based on the embedding silver-colored structure of tetrahedron |
| GB2578871A (en) * | 2018-11-09 | 2020-06-03 | Univ Surrey | Sweat analysis |
| CN109738637A (en) * | 2019-01-17 | 2019-05-10 | 南方医科大学 | A kind of quick detection kit and detection method of skin care item mesocuticle Porcine HGF |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02268744A (en) * | 1989-04-12 | 1990-11-02 | Kawasaki Steel Corp | Method for detecting fingerprint |
| CN101071135A (en) * | 2005-09-26 | 2007-11-14 | 东北师范大学 | Method for preparing semiconductor nano crystal of bio marker utilizing Raman signal |
| CN101268946A (en) * | 2008-04-30 | 2008-09-24 | 东北师范大学 | Method for latency fingerprint appearance of surface functionalization nano-gold particle |
| CN101703404A (en) * | 2009-08-28 | 2010-05-12 | 东北师范大学 | Method for showing fingerprints on various object surfaces and keeping DNA information |
| CN102156119A (en) * | 2011-05-06 | 2011-08-17 | 东北师范大学 | Method for detecting potential information in fingerprints by utilizing Raman spectrum |
| CN102727213A (en) * | 2012-06-20 | 2012-10-17 | 浙江大学 | Method for manifesting latent fingerprints on basis of electrochemical luminescence marker |
-
2013
- 2013-10-18 CN CN201310493627.0A patent/CN103536296B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02268744A (en) * | 1989-04-12 | 1990-11-02 | Kawasaki Steel Corp | Method for detecting fingerprint |
| CN101071135A (en) * | 2005-09-26 | 2007-11-14 | 东北师范大学 | Method for preparing semiconductor nano crystal of bio marker utilizing Raman signal |
| CN101268946A (en) * | 2008-04-30 | 2008-09-24 | 东北师范大学 | Method for latency fingerprint appearance of surface functionalization nano-gold particle |
| CN101703404A (en) * | 2009-08-28 | 2010-05-12 | 东北师范大学 | Method for showing fingerprints on various object surfaces and keeping DNA information |
| CN102156119A (en) * | 2011-05-06 | 2011-08-17 | 东北师范大学 | Method for detecting potential information in fingerprints by utilizing Raman spectrum |
| CN102727213A (en) * | 2012-06-20 | 2012-10-17 | 浙江大学 | Method for manifesting latent fingerprints on basis of electrochemical luminescence marker |
Non-Patent Citations (2)
| Title |
|---|
| 《潜指纹显现方法研究进展》;陈艳 等;《应用化学》;20111031;第28卷(第10期);第1099-1105页 * |
| 《纳米材料在潜指纹显现中的应用》;周小凤 等;《刑事技术》;20130615;第2013卷(第3期);第19-23页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103536296A (en) | 2014-01-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103536296B (en) | Method of detecting human sweat secretion | |
| Wei et al. | Recent advances in the chemical imaging of human fingermarks (a review) | |
| Friesen | Forensic chemistry: The revelation of latent fingerprints | |
| Bumbrah et al. | Emerging latent fingerprint technologies: a review | |
| Sodhi et al. | Physical developer method for detection of latent fingerprints: A review | |
| Qian et al. | Wearable chemosensors: A review of recent progress | |
| CN102727213B (en) | Method for manifesting latent fingerprints on basis of electrochemical luminescence marker | |
| CN104122247B (en) | Glycoprotein detection method based on molecular imprinting technique and Raman spectrum and application | |
| CN103308675B (en) | The preparation of the screen printing electrode immunosensor of quick detection Microcystin and detection method | |
| FR2831789A1 (en) | Process for determining skin type for cosmetic purposes by applying a drop of liquid to the skin and measuring the area wetted by the drop | |
| Lennard | Fingerprint detection: current capabilities | |
| CN102755166B (en) | Method for directly showing latent fingerprints based on multiple electrochemistry lighting systems | |
| Lu et al. | Towards single molecule biosensors using super-resolution fluorescence microscopy | |
| Shi et al. | Label-free physical and electrochemical imaging of latent fingerprints by water and SECM | |
| Azman | Fast, easy, reproducible method for planting fingerprints for ninhydrin, iodine development | |
| Qin et al. | Label‐Free Electrochemical Imaging of Latent Fingerprints on Metal Surfaces | |
| Lin et al. | Surface-enhanced Raman spectroscopic identification in fingerprints based on adhesive Au nanofilm | |
| Sciutto et al. | Localization of proteins in paint cross-sections by scanning electrochemical microscopy as an alternative immunochemical detection technique | |
| Zalaffi et al. | electrochemical and SERS sensors for cultural heritage diagnostics and conservation: recent advances and prospects | |
| Markina et al. | Nanoparticle-based paper sensor for thiols evaluation in human skin | |
| Tian et al. | Direct detection of label-free blood fingermarks by SECM imaging | |
| CN103536295A (en) | Potential fingerprint imaging method based on electrochemiluminescence immunoassay | |
| Sharma et al. | Preparation of cotton fabric based non-invasive colorimetric sensor for instant detection of ketones | |
| Fan et al. | Dye-soaked cotton pads for latent blood fingerprint development | |
| Wei et al. | Colorimetric visualization and SECM imaging of latent fingerprints on food surfaces |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |