CN105295907A - Preparation method of functional rare earth long-afterglow nanocomposite and latent fingerprint imaging application of functional rare earth long-afterglow nanocomposite - Google Patents
Preparation method of functional rare earth long-afterglow nanocomposite and latent fingerprint imaging application of functional rare earth long-afterglow nanocomposite Download PDFInfo
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- CN105295907A CN105295907A CN201510697012.9A CN201510697012A CN105295907A CN 105295907 A CN105295907 A CN 105295907A CN 201510697012 A CN201510697012 A CN 201510697012A CN 105295907 A CN105295907 A CN 105295907A
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Abstract
The invention discloses a preparation method of functional rare earth long-afterglow nanocomposite and latent fingerprint imaging application of the functional rare earth long-afterglow nanocomposite and belongs to the technical field of biological analysis and fingerprint detection. Avidin is modified on the surface of an amino-functionalized strontium aluminate rare earth doped long-afterglow nano material, an avidin-functionalized rare earth long-afterglow nanocomposite is prepared, and therefore the avidin-functionalized rare earth long-afterglow nanocomposite is connected to a latent fingerprint marked by a biotinylated antibody through the recognition effect between biotin and avidin. The strontium aluminate rare earth doped long-afterglow nano material has the long-afterglow characteristic and still keeps emitting light after light source irradiation and light source removal, and latent fingerprint imaging can be achieved. The method is fast, simple and free of background interference.
Description
Technical field
The invention discloses a kind of preparation method of functional rare earth long-persistence nano matrix material and the application in laten fingerprints imaging thereof, belong to bioanalysis and fingerprinting detection technique field.
Background technology
Laten fingerprints is the fingerprint lines formed as sweat transfer through health nature secretory product, and visually not easily finding, is modal fingerprint in spot.According to China's various places incomplete statistics, in the criminal case of police's detection, utilize fingerprint to solve a case and account for more than seventy percent, and ratio rises year by year.Under normal circumstances, fingerprint is the most direct material evidence that offender stays at the scene, so the appearing technique of research latent fingerprint improves the taking rate of fingerprint material evidence and the important step of utilization ratio.
The development of modern dactylography has had the history of upper a century, and according to the difference manifesting principle, traditional latent fingerprint process for show is mainly divided three classes: optics appearance method, chemical appearance method, physisorphtion.Its ultimate principle is the sweat material using a kind of light or material to act on finger mark, makes the latent fingerprint that can not see become visible.Be developed so far, fingerprint detection not only at forensic identification, individual recognition important role, is widely used in the fields such as the safety verification in daily life, access control, personal authentication simultaneously.In recent years along with the development of analysis science, people not only rest on the research of fingerprint and utilize conventional physical or chemical means to observe its pattern, more be devoted to develop various emerging technology (as mass spectrum imaging, infrared Raman imaging, scan-type electrochemical microscope and fluorescence immunoassay imaging etc.) simultaneously, analyzing and testing is carried out to the composition in fingerprint, thus excavates out more how valuable biology and medical information.As, whether the whether contacted explosive substance of terrorist, a people have smoking habit etc., and scientist can obtain relevant information by fingerprint detection.Although Appearing Techniques of Human Fingerprints plays significant role in criminal investigation and analysis science, still there is many difficulties in fingerprint manifestation field.As, in traditional method, mostly will quote a kind of allogenic material and manifest latent fingerprint, treating processes is more loaded down with trivial details, and has destructiveness to sample.Certain somatic damage is there is in the small grain size powder adopted in now widely used fumigation and brush dust method to technician.And emerging fingerprint is as mass spectrum imaging method and infrared Raman imaging method, then needs the operative knowledge using expensive large-scale instrument and specialty, be unfavorable for popularizing.
Rare earth ion doped long after glow luminous material has good light stability, wider excitation spectrum and narrower emmission spectrum, excitation spectrum and is separated with emmission spectrum and the optical signature such as tunable emmission spectrum wavelength, overcome organic fluorescent powder easily by many defects such as photobleaching, emmission spectrum be wider, especially its overlength afterglow characteristic gets more and more people's extensive concerning.Rare earth ion doped long after glow luminous material has long afterglow property, can absorb and store luminous energy, and still can be luminous for a long time after removing light source, the present invention has prepared functional rare earth long after glow luminous material, and the long afterglow property of rare earth long-afterglow luminescent material is combined with immunological technique, be applied to the laten fingerprints imaging on common impermeability object and complex background object, for realizing, laten fingerprints imaging, effectively elimination background interference etc. are significant.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of functional rare earth long-persistence nano matrix material, utilize the effect between vitamin H and avidin, the strontium aluminate rare earth of avidin doping long-persistence nano matrix material is attached to the laten fingerprints surface that biotinylated antibody is modified, this fingerprint is through illumination and after removing light source, still can keep luminous and realize laten fingerprints imaging, it has fast, simply, without the feature of background interference.
The present invention realizes like this, first by teos hydrolysis at strontium aluminate rare earth doping long-persistence nano composite material surface coated silica layer, add 3-aminopropyl triethoxysilane again and make its surface amination, then utilize glutaraldehyde avidin to be connected to amination strontium aluminate rare earth doping long-persistence nano composite material surface, prepare avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material; Utilize the recognition reaction between vitamin H and avidin, avidin strontium aluminate rare earth doping long-persistence nano matrix material is attached to the laten fingerprints surface that marked biotinylated antibody, utilize the long afterglow property of strontium aluminate rare earth doping long-persistence nano material, fingerprint is through illumination and after removing light source, still can keep luminous, thus achieve fast, simple, without the laten fingerprints imaging of background interference.
The present invention is by the following technical solutions:
(1) preparation of biotin functionalized strontium aluminate rare earth doping long-persistence nano matrix material:
1) strontium aluminate rare earth is adulterated long-persistence nano material at 3000rpm centrifuging and taking supernatant liquor, grind after vacuum-drying, get the ultrasonic mixing in dehydrated alcohol of 1.6mg strontium aluminate rare earth doping long-persistence nano dispersion of materials, add the mixing solutions of 222 μ L ethanol, 247 μ L deionized waters, 6.7 μ L tetraethoxys, ultrasonic 5min, add 25 μ L ammoniacal liquor (28% ~ 30%), ultrasonic 30min, 1200rpm magnetic agitation 7.5h.Centrifugal rear washing with alcohol 3 times, 36 ° of C vacuum-dryings, make the strontium aluminate rare earth doping long-persistence nano material of coated with silica.
2) by dispersion of materials prepared in step 1) at 2mLN, ultrasonic mixing in dinethylformamide (DMF), add 20 μ L3-aminopropyl triethoxysilanes, 1300rpm magnetic agitation 5h, 3 times are washed respectively with DMF, ethanol, 36 ° of C vacuum-dryings, make amination strontium aluminate rare earth doping long-persistence nano material.
3) by step 2) in prepared dispersion of materials in phosphoric acid salt (PBS) buffered soln of the pH7.4 containing 5% glutaraldehyde, 900rpm magnetic agitation 1h, with the PBS buffered soln centrifuge washing 3 times of pH7.4, precipitation is scattered in 1mLPBS buffered soln, add the avidin of 400 μ L1mg/mL, 4 ° of C react 12h, make avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material.
(2) the laten fingerprints imaging applications of biotin functionalized strontium aluminate rare earth doping long-persistence nano matrix material: gather fingerprint on the glass sheet, containing N,O-Diacetylmuramidase on fingerprint ridge line, utilize the specific recognition effect of antigen/antibody, anti-for rabbit N,O-Diacetylmuramidase polyclonal antibody is modified on fingerprint; By the anti-N,O-Diacetylmuramidase polyclonal antibody of rabbit to the specific binding of donkey anti-rabbit IgG, biotin labeled donkey anti-rabbit IgG is connected to fingerprint surface; And then utilize vitamin H and avidin effect, avidin functionalization strontium aluminate rare earth long afterglow nano composite material is attached to fingerprint surface.Remove light source after illumination, use collected by camera finger printing, realize laten fingerprints imaging.
Technique effect of the present invention is: avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material is modified the laten fingerprints surface being marked with biotinylated antibody, utilize the long afterglow property of strontium aluminate rare earth doping long-afterglow material, through illumination and after removing light source, strontium aluminate rare earth doping long-persistence nano matrix material still keeps luminous, thus realizes laten fingerprints imaging.Method of the present invention has fast, simple, without features such as background interference.
Accompanying drawing explanation
Fig. 1 is that (a) functionalization strontium aluminate rare earth doping long-afterglow material preparation and (b) laten fingerprints are modified and imaging schematic diagram.
Fig. 2 is strontium aluminate rare earth doping long-persistence nano material (a) fluorescent exciting spectrogram and (b) fluorescence emission spectrogram.
Fig. 3 is the decay of afterglow curve of strontium aluminate rare earth doping long-persistence nano material.
Fig. 4 is Fourier transform infrared spectroscopy figure: (a) strontium aluminate rare earth doping long-persistence nano material, b () strontium aluminate rare earth doping long-persistence nano material coated silica, (c) is the material after (b) further amination.
Fig. 5 is laten fingerprints collection of illustrative plates: (a) fingerprint, and (b) fingerprint/primary antibodie/biotinylation two resists, and (c) fingerprint/primary antibodie/biotinylation two resists/avidin strontium aluminate rare earth doping long-persistence nano material.
Fig. 6 is there to be the waterproof paper of fluorescence to compare for the laten fingerprints picture of substrate gained as stated above under ultraviolet lamp: (a) is after under ultra violet lamp, (b) turns off ultraviolet lamp.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is further elaborated, and the present invention is not limited to this;
embodiment 1
By the biotin functionalized strontium aluminate rare earth doping of preparation schematic diagram (a) in Fig. 1 Suo Shi long-persistence nano matrix material:
(1) strontium aluminate rare earth is adulterated long-persistence nano material at 3000rpm centrifuging and taking supernatant liquor, grind after vacuum-drying, get the ultrasonic mixing in dehydrated alcohol of 1.6mg strontium aluminate rare earth doping long-persistence nano dispersion of materials, add the mixing solutions of 222 μ L ethanol, 247 μ L deionized waters, 6.7 μ L tetraethoxys, ultrasonic 5min, add 25 μ L ammoniacal liquor (28% ~ 30%), ultrasonic 30min, 1200rpm magnetic agitation 7.5h.Centrifugal rear washing with alcohol 3 times, 36 ° of C vacuum-dryings, make the strontium aluminate rare earth doping long-persistence nano material of coated with silica.
(2) by dispersion of materials prepared in step (1) at 2mLN, ultrasonic mixing in dinethylformamide (DMF), add 20 μ L3-aminopropyl triethoxysilanes, 1300rpm magnetic agitation 5h, 3 times are washed respectively with DMF, ethanol, 36 ° of C vacuum-dryings, make amination strontium aluminate rare earth doping long-persistence nano material.
(3) by dispersion of materials prepared in step (2) in phosphoric acid salt (PBS) buffered soln of the pH7.4 containing 5% glutaraldehyde, 900rpm magnetic agitation 1h, with the PBS buffered soln centrifuge washing 3 times of pH7.4, precipitation is scattered in 1mLPBS buffered soln, add the avidin of 400 μ L1mg/mL, 4 ° of C react 12h, make avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material.
Fig. 2 is strontium aluminate rare earth doping long-persistence nano material (a) exciting light spectrogram and (b) fluorescence emission spectrogram.As seen from the figure, this material has good luminescent properties, optimum excitation wave is about 360nm, optimum transmit wavelength is about 530nm, and excitation wavelength range is wide, even can be extended to visible region, show that not only ultraviolet lamp can excite and visible ray also can realize exciting, so simple and practicality makes to have a good application prospect.
Fig. 3 is the decay of afterglow curve of strontium aluminate rare earth doping long-persistence nano material.Through illumination and after removing light source for some time, light intensity still can distinguish, there is good practicality.Because strontium aluminate rare earth doping long-persistence nano material can have good long afterglow property with day optical excitation, much have the material of endogenous fluorescence to cause interference, visible, this material is used for laten fingerprints imaging without background interference.
Adopt Fourier transform infrared spectroscopy to characterize the process that silicon-dioxide and 3-aminopropyl triethoxysilane modify strontium aluminate rare earth doping long-persistence nano material, result as shown in Figure 4.Wherein, a () is strontium aluminate rare earth doping long-persistence nano material, b () is coated with silica strontium aluminate rare earth doping long-persistence nano material, (c) for after coated silica again through 3-aminopropyl triethoxysilane amidized strontium aluminate rare earth doping long-persistence nano material.Adulterate compared with long-persistence nano material with strontium aluminate rare earth, the strontium aluminate rare earth doping long-persistence nano material after coated silicon-dioxide is at 1165cm
-1and 845cm
-1having there is the asymmetric stretching vibration of Si-O-Si and Si-O-Si symmetrical stretching vibration in place, shows respectively at the coated silicon-dioxide of strontium aluminate rare earth doping long-persistence nano material surface success.Strontium aluminate rare earth doping long-persistence nano material after coated silicon-dioxide again through the amination of 3-aminopropyl triethoxysilane, at 2980 ~ 2900cm
-1the new absorption peak that place occurs corresponds to the alkane-CH of 3-aminopropyl triethoxysilane
2, C-H stretching vibration peak, showing that 3-aminopropyl triethoxysilane has successfully modified coated with silica strontium aluminate rare earth doping long-persistence nano material surface, providing abundant amino for connecting avidin further by glutaraldehyde.
embodiment 2
The laten fingerprints imaging applications of biotin functionalized strontium aluminate rare earth doping long-persistence nano matrix material: gather fingerprint on the glass sheet, anti-for the certain density rabbit of 100 μ L N,O-Diacetylmuramidase polyclonal antibody is dripped on laten fingerprints, incubated at room 30min, by the specific recognition effect of antigen/antibody, the N,O-Diacetylmuramidase of the anti-N,O-Diacetylmuramidase polyclonal antibody of rabbit on fingerprint ridge line is combined, sample fingerprint after hatching is dipped in the PBS buffered soln of pH7.4 and jiggles cleaning twice, the antibody of removing non-specific binding, argon gas dries up.Drip certain density biotin labeled donkey anti-rabbit IgG100 μ L on fingerprint surface, hatch 30min, by the anti-N,O-Diacetylmuramidase polyclonal antibody of rabbit to the specific binding of donkey anti-rabbit IgG, biotin labeled donkey anti-rabbit IgG is connected to fingerprint surface, with PBS cleaning, argon gas dries up.100 μ L certain density avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material is dripped on fingerprint surface, biotin-labeled pentylamine is made to react 30min under room temperature, recognition reaction between vitamin H and avidin makes avidin functionalization strontium aluminate rare earth long afterglow nano composite material be attached to fingerprint surface, wash away unnecessary particle with PBS, dry up with argon gas.Remove light source after illumination 5min, use collected by camera finger printing under dark surrounds, thus realize laten fingerprints imaging, result as shown in Figure 5.From the laten fingerprints imaging comparison diagram of Fig. 5, simple fingerprint and fingerprint/antibody cannot realize laten fingerprints imaging, only have when avidin strontium aluminate rare earth doping long-persistence nano material being attached to fingerprint surface by biotin-avidin effect and could obtain laten fingerprints image clearly.
In order to embody the advantage that avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material is applied in laten fingerprints imaging further, we are to have the waterproof paper of fluorescence background for substrate, carry out laten fingerprints imaging according to the method described above, as seen from Figure 6, under ultra violet lamp, the fluorescence background of waterproof paper is very strong, cannot differentiate the strontium aluminate rare earth doping long-persistence nano matrix material be modified on laten fingerprints, so can not realize laten fingerprints imaging.But after removing ultraviolet lamp, in dark surrounds, waterproof paper unstressed configuration, the luminescence of strontium aluminate rare earth doping long-persistence nano matrix material is then visible, thus obtains laten fingerprints image clearly.Visible, the inventive method can eliminate background fluorescence well, realizes the laten fingerprints imaging without background interference, has good application potential.
Claims (3)
1. the preparation method of functional rare earth long-persistence nano matrix material, it is characterized in that: by teos hydrolysis at strontium aluminate rare earth doping long-persistence nano composite material surface coated silica layer, add 3-aminopropyl triethoxysilane and make its amination, utilize glutaraldehyde avidin to be connected to amination strontium aluminate rare earth doping long-persistence nano composite material surface, be prepared into avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material.
2. the preparation method of functional rare earth long-persistence nano matrix material according to claim 1, is characterized in that: concrete steps are:
1) strontium aluminate rare earth is adulterated long-persistence nano material at 3000rpm centrifuging and taking supernatant liquor, grind after vacuum-drying, get the ultrasonic mixing in dehydrated alcohol of 1.6mg strontium aluminate rare earth doping long-persistence nano dispersion of materials, add the mixing solutions of 222 μ L ethanol, 247 μ L deionized waters and 6.7 μ L tetraethoxys, ultrasonic 5min, add 25 μ L ammoniacal liquor (28% ~ 30%), ultrasonic 30min, 1200rpm magnetic agitation 7.5h; Centrifugal rear washing with alcohol 3 times, 36 ° of C vacuum-dryings, make the strontium aluminate rare earth doping long-persistence nano material of coated with silica;
2) by dispersion of materials prepared in step 1) at 2mLN, ultrasonic mixing in dinethylformamide, add 20 μ L3-aminopropyl triethoxysilanes, 1300rpm magnetic agitation 5h, with N, dinethylformamide, ethanol wash 3 times respectively, 36 ° of C vacuum-dryings, make amination strontium aluminate rare earth doping long-persistence nano material;
3) by step 2) in prepared dispersion of materials in the phosphate buffer soln of the pH7.4 containing 5% glutaraldehyde, 900rpm magnetic agitation 1h, with the phosphate buffer soln centrifuge washing 3 times of pH7.4, precipitation is scattered in 1mL phosphate buffer soln, add the avidin of 400 μ L1mg/mL, 4 ° of C react 12h, make avidin functionalization strontium aluminate rare earth doping long-persistence nano matrix material.
3. the laten fingerprints imaging applications of functional rare earth long-persistence nano matrix material, it is characterized in that: gather fingerprint on the glass sheet, containing N,O-Diacetylmuramidase on fingerprint ridge line, utilize the specific recognition effect of antigen/antibody, anti-for rabbit N,O-Diacetylmuramidase polyclonal antibody is modified on fingerprint; By the anti-N,O-Diacetylmuramidase polyclonal antibody of rabbit to the specific binding of donkey anti-rabbit IgG, biotin labeled donkey anti-rabbit IgG is connected to fingerprint surface; And then utilize vitamin H and avidin effect, the avidin functionalization strontium aluminate rare earth long afterglow nano composite material prepared by claim 1 or 2 is attached on fingerprint; After removing light source, use collected by camera finger printing after illumination, laten fingerprints imaging can be realized.
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