CN105372420A - Method for detecting ancient wool fabric by microscope - Google Patents

Method for detecting ancient wool fabric by microscope Download PDF

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Publication number
CN105372420A
CN105372420A CN201510768133.8A CN201510768133A CN105372420A CN 105372420 A CN105372420 A CN 105372420A CN 201510768133 A CN201510768133 A CN 201510768133A CN 105372420 A CN105372420 A CN 105372420A
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China
Prior art keywords
vessels
pbs
carrying
punching
relic sample
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CN201510768133.8A
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胡智文
刘建军
王秉
游秋实
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Zhejiang University of Technology ZJUT
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Zhejiang University of Technology ZJUT
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Priority to CN201510768133.8A priority Critical patent/CN105372420A/en
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Treatment Of Fiber Materials (AREA)
  • Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

The invention discloses a method for detecting an ancient wool fabric by a microscope. The method comprises A, preparing a PBS 7.4 buffer solution, B, drawing three single fibers from a cultural relic sample, respectively arranging the single fibers in punching vessels marked as 1, 2, and 3, carrying out washing through the PBS buffer solution so that attachments on the surface are removed completely, adding a rabbit anti-keratin polyclonal antibody into the vessel 1, adding the PBS buffer solutions into the vessels 2 and 3, putting the vessels into a refrigerator, carrying out standing overnight, respectively adding the PBS buffer solutions into the punching vessels and carrying out immersion and washing, C, respectively adding colloidal gold-labeled goat anti-rabbit IgG(H+L) antibodies into the vessels 2 and 1, adding the PBS solution into the vessel 3, respectively putting the vessels 1, 2 and 3 into a box with a wet towel pad, carrying out incubation, respectively adding the PBS solutions into the punching vessels, carrying out immersion and washing and carrying sealing by cover glasses, and D, observing the vessels 1, 2 and 3 through a microscope, and comparing the detection results, wherein when colors of the materials in the vessels 2 and 3 do not change and the color of the material in the vessel 1 has a bright red color, it is proved that the cultural relic sample is specifically bonded to the rabbit anti-keratin antibody and thus the cultural relic sample is a wool fabric.

Description

A kind of method utilizing microscopic examination woollen fabrics in ancient times
Technical field
The present invention relates to a kind of detection method utilizing microscopic examination woollen fabrics, be mainly used in the detection of woollen fabrics in ancient times.
Background technology
Ancient times are widely used in dress ornament and woollen blanket at northern area woolen knitwear, and wool fabric accounts for the overwhelming majority.The silk relics of its luxury aspect of a large amount of wool fabrics be unearthed not second to being unearthed, this is not only the important tangible evidence of ancient textile change of technique, also has important references to research ancient society environment and humanity activities and is worth.And the further investigation lacked in prior art wool keratin qualification in Ancient Silk Textile.
Wool keratin is a kind of sclerosis, not diffluent protein, wherein existing containing carboxyl etc. at interior acidic groups, also have amino-contained, imido grpups etc. are at interior basic group, so wool all has certain instability in acid, alkali, salt, oxygenant, reductive agent, halogen etc.In silk fabric cultural relics qualification, this causes certain difficulty to the discriminating of wool product.And Elisa method qualification wool silk goods are quick, convenient, woollen fabrics can be identified by efficiently and accurately, thus provide foundation for woollen fabrics developmental research in ancient times.
Summary of the invention
The technical problem to be solved in the present invention is: provide one intuitively for above-mentioned prior art Problems existing, utilize the method for microscopic examination woollen fabrics in ancient times efficiently, first rabbit anti-keratin polyclonal antibody be impregnated in historical relic sample surface, after washes clean, add goat antirabbit lgG (H+L) antibody of colloid gold label, form the anti-two anti-compounds of antigen-one, then historical relic sample is placed on basis of microscopic observation by washes clean, and whether surface presents redness and carry out qualitative analysis per sample.
For technical solution problem, the technical scheme taked is: the method utilizing microscopic examination woollen fabrics in ancient times, it is characterized in that adopting following steps:
A) PBS7.4 buffer: KCl0.2g, KH 2pO 40.27g, NaCl8.0g, Na 2hPO 412H 2o3.58g, is settled to 1000mL with distilled water.
B) inside historical relic sample, extract three single fibers, be positioned over respectively in punching ware, be labeled as 1., 2., 3.; Wash with the PBS buffer solution of PH=7.4, ensure surface attachments washes clean; 80-120 μ l PBS7.4 solution dilution 10-50 rabbit anti-keratin polyclonal antibody is doubly added in 1.; 2., add the PBS damping fluid of 80-120 μ lPH=7.4 3., be then positioned over refrigerator, 3-5 DEG C is spent the night; Then the PBS solution adding 1-3mlPH=7.4 respectively, in punching ware, is soaked 4-6min and is washed, wash 2-4 time;
C) respectively to 1., add 80-120 μ l PBS7.4 solution dilution 100-400 colloid gold label goat antirabbit lgG (H+L) antibody doubly 2., 3., add the PBS solution of 80-120 μ lPH=7.4; Then respectively will 1., 2., be 3. positioned in box that wet towel cushions, hatch 1-2h for 37-39 DEG C; Then the PBS solution adding 1-3mlPH=7.4 respectively, in punching ware, is soaked 4-6min and is washed, wash 2-4 time; Use cover glass mounting again;
D) will 1., 2., observe under being 3. positioned over microscope; , 3. there is not color change, and 1. present brighter redness, illustrate that historical relic sample and rabbit antikeratin antibody there occurs specific binding, prove that historical relic sample is woollen fabrics in the result relatively recorded: 2..
Useful achievement of the present invention is: the present invention adopts immune colloidal gold technique to detect woollen fabrics in ancient times, and compared to existing technology, the method simple and fast, highly sensitive, high specificity, visual result, is more suitable for archeological site analysis.
Embodiment
Below embodiments of the invention are elaborated.Following examples are only further described the application, should not be construed as the restriction to the application.
Embodiment 1:
Utilize a detection method for microscopic examination woollen fabrics, adopt following steps:
A) PBS7.4 buffer: KCl0.2g, KH2PO40.27g, NaCl8.0g, Na2HPO412H2O3.58g, be settled to 1000mL with distilled water.
B) inside historical relic sample, extract three single fibers, be positioned over respectively in punching ware, be labeled as 1., 2., 3.; Wash with the PBS buffer solution of PH=7.4, ensure surface attachments washes clean.Respectively to the rabbit anti-keratin polyclonal antibody adding 80 μ l PBS7.4 solution dilution 10 times in 1.; 2., add the PBS damping fluid of 80 μ lPH=7.4 3., be then positioned over refrigerator, 4 DEG C are spent the night.Then the PBS solution adding 2mlPH=7.4 respectively, in punching ware, is soaked 5min and is washed, wash 3 times;
C) respectively to 1., add colloid gold label goat antirabbit lgG (H+L) antibody of 80 μ l PBS7.4 solution dilution 100 times 2., 3., add the PBS solution of 80 μ lPH=7.4; Then respectively will 1., 2., be 3. positioned in box that wet towel cushions, hatch 1h for 37 DEG C; Then the PBS solution adding 2mlPH=7.4 respectively, in punching ware, is soaked 5min and is washed, wash 2 times; Use cover glass mounting again;
D) will 1., 2., observe under being 3. positioned over microscope; , 3. there is not color change, and 1. present brighter redness, illustrate that historical relic sample and rabbit antikeratin antibody there occurs specific binding, prove that historical relic sample is woollen fabrics in the result relatively recorded: 2..
Embodiment 2
Utilize a detection method for microscopic examination woollen fabrics, adopt following steps:
A) PBS7.4 buffer: KCl0.2g, KH2PO40.27g, NaCl8.0g, Na2HPO412H2O3.58g, be settled to 1000mL with distilled water.
B) inside historical relic sample, extract three single fibers, be positioned over respectively in punching ware, be labeled as 1., 2., 3.; Wash with the PBS buffer solution of PH=7.4, ensure surface attachments washes clean.Respectively to the rabbit anti-keratin polyclonal antibody adding 100 μ l PBS7.4 solution dilution 25 times in 1.; 2., add the PBS damping fluid of 100 μ lPH=7.4 3., be then positioned over refrigerator, 3 DEG C are spent the night.Then the PBS solution adding 1mlPH=7.4 respectively, in punching ware, is soaked 4min and is washed, wash 2 times;
C) respectively to 1., add colloid gold label goat antirabbit lgG (H+L) antibody of 100 μ l PBS7.4 solution dilution 200 times 2., 3., add the PBS solution of 100 μ lPH=7.4; Then respectively will 1., 2., be 3. positioned in box that wet towel cushions, hatch 1h for 38 DEG C; Then the PBS solution adding 1mlPH=7.4 respectively, in punching ware, is soaked 5min and is washed, wash 3 times; Use cover glass mounting again;
D) will 1., 2., observe under being 3. positioned over microscope; , 3. there is not color change, and 1. present brighter redness, illustrate that historical relic sample and rabbit antikeratin antibody there occurs specific binding, prove that historical relic sample is woollen fabrics in the result relatively recorded: 2..
Embodiment 3
Utilize a detection method for microscopic examination woollen fabrics, adopt following steps:
A) PBS7.4 buffer: KCl0.2g, KH2PO40.27g, NaCl8.0g, Na2HPO412H2O3.58g, be settled to 1000mL with distilled water.
B) inside historical relic sample, extract three single fibers, be positioned over respectively in punching ware, be labeled as 1., 2., 3.; Wash with the PBS buffer solution of PH=7.4, ensure surface attachments washes clean.Respectively to the rabbit anti-keratin polyclonal antibody adding 120 μ l PBS7.4 solution dilution 50 times in 1.; 2., add the PBS damping fluid of 120 μ lPH=7.4 3., be then positioned over refrigerator, 5 DEG C are spent the night.Then the PBS solution adding 3mlPH=7.4 respectively, in punching ware, is soaked 6min and is washed, wash 4 times;
C) respectively to 1., add colloid gold label goat antirabbit lgG (H+L) antibody of 120 μ l PBS7.4 solution dilution 400 times 2., 3., add the PBS solution of 120 μ lPH=7.4; Then respectively will 1., 2., be 3. positioned in box that wet towel cushions, hatch 1h for 39 DEG C; Then the PBS solution adding 2mlPH=7.4 respectively, in punching ware, is soaked 5min and is washed, wash 4 times; Use cover glass mounting again;
D) will 1., 2., observe under being 3. positioned over microscope; , 3. there is not color change, and 1. present brighter redness, illustrate that historical relic sample and rabbit antikeratin antibody there occurs specific binding, prove that historical relic sample is woollen fabrics in the result relatively recorded: 2..

Claims (1)

1. utilize a method for microscopic examination woollen fabrics in ancient times, it is characterized in that comprising the steps:
A) PBS7.4 buffer: KCl0.2g, KH 2pO 40.27g, NaCl8.0g, Na 2hPO 412H 2o3.58g, is settled to 1000mL with distilled water.
B) inside historical relic sample, extract three single fibers, be positioned over respectively in punching ware, be labeled as 1., 2., 3.; Wash with the PBS buffer solution of PH=7.4, ensure surface attachments washes clean; 80-120 μ l PBS7.4 solution dilution 10-50 rabbit anti-keratin polyclonal antibody is doubly added in 1.; 2., add the PBS damping fluid of 80-120 μ lPH=7.4 3., be then positioned over refrigerator, 3-5 DEG C is spent the night; Then the PBS solution adding 1-3mlPH=7.4 respectively, in punching ware, is soaked 4-6min and is washed, wash 2-4 time;
C) respectively to 1., add 80-120 μ l PBS7.4 solution dilution 100-400 colloid gold label goat antirabbit lgG (H+L) antibody doubly 2., 3., add the PBS solution of 80-120 μ lPH=7.4; Then respectively will 1., 2., be 3. positioned in box that wet towel cushions, hatch 1-2h for 37-39 DEG C; Then the PBS solution adding 1-3mlPH=7.4 respectively, in punching ware, is soaked 4-6min and is washed, wash 2-4 time; Use cover glass mounting again;
D) will 1., 2., observe under being 3. positioned over microscope; , 3. there is not color change, and 1. present brighter redness, illustrate that historical relic sample and rabbit antikeratin antibody there occurs specific binding, prove that historical relic sample is woollen fabrics in the result relatively recorded: 2..
CN201510768133.8A 2015-11-12 2015-11-12 Method for detecting ancient wool fabric by microscope Pending CN105372420A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106596970A (en) * 2016-12-12 2017-04-26 浙江理工大学 Method for measuring ancient cowhair micro-trace based on proteomics

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6730493B1 (en) * 1998-08-04 2004-05-04 Wella Aktiengesellschaft Method for determining the state of substances containing keratin and suitable devices and means therefor
JP2007240180A (en) * 2006-03-06 2007-09-20 Mandom Corp Damage evaluation method of keratin fiber
EP1847831A1 (en) * 2006-04-21 2007-10-24 Consiglio Nazionale delle Ricerche Method for the determination of the composition of textile fibers
CN104406992A (en) * 2014-12-31 2015-03-11 浙江理工大学 Method for detecting ancient silk fabrics by transmission electron microscope
CN104459104A (en) * 2014-12-31 2015-03-25 浙江理工大学 Method for detecting antique silk fabric through microscope
CN104483478A (en) * 2014-12-31 2015-04-01 浙江理工大学 Detection method for ancient silk fabrics

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6730493B1 (en) * 1998-08-04 2004-05-04 Wella Aktiengesellschaft Method for determining the state of substances containing keratin and suitable devices and means therefor
JP2007240180A (en) * 2006-03-06 2007-09-20 Mandom Corp Damage evaluation method of keratin fiber
EP1847831A1 (en) * 2006-04-21 2007-10-24 Consiglio Nazionale delle Ricerche Method for the determination of the composition of textile fibers
CN104406992A (en) * 2014-12-31 2015-03-11 浙江理工大学 Method for detecting ancient silk fabrics by transmission electron microscope
CN104459104A (en) * 2014-12-31 2015-03-25 浙江理工大学 Method for detecting antique silk fabric through microscope
CN104483478A (en) * 2014-12-31 2015-04-01 浙江理工大学 Detection method for ancient silk fabrics

Non-Patent Citations (2)

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Title
CAROLIN SOLAZZO，等: "Modeling deamidation in sheep α-keratin peptides and applicaiton to archelolgical wool textiles", 《ANALYTICAL CHEMISTRY》 *
俞宁，等: "小河墓地出土羊毛文物检测及其老化过程的人工模拟", 《浙江理工大学学报(自然科学版)》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106596970A (en) * 2016-12-12 2017-04-26 浙江理工大学 Method for measuring ancient cowhair micro-trace based on proteomics

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