CN104142322A - Method for quickly authenticating nature of peripheral nerve by raman spectra technique and dyeing - Google Patents
Method for quickly authenticating nature of peripheral nerve by raman spectra technique and dyeing Download PDFInfo
- Publication number
- CN104142322A CN104142322A CN201410405809.2A CN201410405809A CN104142322A CN 104142322 A CN104142322 A CN 104142322A CN 201410405809 A CN201410405809 A CN 201410405809A CN 104142322 A CN104142322 A CN 104142322A
- Authority
- CN
- China
- Prior art keywords
- raman
- nerve
- dyeing
- fibre
- micro
- 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.)
- Pending
Links
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to a method for quickly authenticating the nature of a peripheral nerve by raman spectra technique and dyeing. The method comprises the following steps of obtaining a fracture surface specimen, which is 1 to 2mm thick, of the peripheral nerve; performing frozen sectioning, wherein each section is 20 to 30 microns thick; under a microscopes raman spectrometer, focusing a laser beam to the fracture surface of a nerve fiber for raman spectrum acquisition; performing Karnovsky-Roots dyeing for 30min; under the microscopes raman spectrometer again, focusing the laser beam with the same wavelength to the fracture surface of the nerve fiber for raman spectrum acquisition; and judging the nature of the nerve fiber according to spectrum results before and after dyeing. According to the method for authenticating the nature of the peripheral nerve, the natures of a moving fiber and a feeling fiber in the peripheral nerve can be quickly identified; the method is relatively high in sensitivity, specificity and accuracy, and various requirements for authenticating the two nerve fibers can be met clinically and on a scientific research.
Description
Technical field
The present invention relates to optical analysis technical field, specifically, is a kind of method of utilizing Raman spectroscopy combined staining to differentiate fast peripheral nerve character.
Background technology
Peripheral nerve injury is comparatively common at Orthopedic Clinical, and the reparation after its damage is the problem of Chinese scholars primary study with regeneration always.As everyone knows, the undesirable main cause of neurological functional recovery is that misorientation between motion or Sensory nerve fibre coincide.It is the most effective restorative procedure of complete dialysis of generally acknowledging that end terminal nerve coincide, simple peripheral nerve injury can be by coincideing restore funcitons without stump under tension force, but to long distance is damaged, cannot use the technology such as allograft that need to use autologous nerve and go antigenize to process.No matter how which kind of repair mode, identify fast and accurately to the character of peripheral nerve fiber, is one of current problem in the urgent need to address, for the selection of modus operandi in peripheral nerve injury art clinically, has good reference value.
The method of identifying clinically at present neural character mainly can be classified as following a few class: 1) according to detecting judgement in anatomic characteristic and art; 2) galvanism in art; 3) cholinesterase or carbonic anhydrase dyeing; 4) isotope method is measured acetyl group choline transferase active etc.These methods are general consuming time longer, can not meet operation traditional Chinese physician and the signal of neural character be carried out to the requirement of quick obtaining and discriminating.
Chinese patent literature CN101246159B discloses a kind of method of quick discriminating peripheral nerve bundle nature, step is: 1. cut the fresh nerve that a segment length is less than or equal to 2mm, put into concentration and be more than or equal to 90% ethanol and soak 10 ~ 20 minutes, then put into concentration and be 20 ~ 40% sucrose solution and soak 10-20 minute; 2. under the condition of-20 ~-40 ℃, on freezing microtome, cut into slices, slice thickness is 20 ~ 50 μ m; 3. utilize the method for coherent imaging, three-dimensional structure to section sample is done coherent imaging, according to tripleplane's feature of imaging sample, carry out the judgement of nerve tract character, wherein the nerve tract of staggered striped area occupied ratio 70 ~ 100% is motor tract, otherwise is sensory nerve bundle.Chinese patent literature CN101246141B discloses a kind of electrochemical detection method of quick discriminating peripheral nerve bundle nature, first adopt polymkeric substance-golden nanometer particle compound modified electrode, then in choline oxidase and acetylcholinesterase solution, hatch successively, by the complexes membrane absorption affinity good to protein, make two kinds of enzymes in electrode surface enrichment, pass through three-electrode system, based on dual-enzyme system, measure the ampere response of substrate acetylcholine, thereby indirectly realize the detection of acetylcholinesterase (detectable concentration scope: 1.0-2.0unit/mL).With nervous homogenates, replace standard acetylcholinesterase solution to hatch, by whether acetylcholine being had to the detection of response, realized the quick discriminating to motor tract and sensory tract.The invention provides a kind of method that has no at present the quick discriminating peripheral nerve character of report.
Summary of the invention
The object of the invention is for deficiency of the prior art, a kind of method of utilizing Raman spectroscopy combined staining to differentiate fast peripheral nerve character is provided.
Research shows, Karnovsky-Roots dyeing is in the time of 1 hour, and it is painted that light the dying of Remak's nerve fiber and motion myelinated fibre almost do not have, and cannot distinguish nerve fibre character; Within 2 hours, starting has light colored, and the nerve fibre quantity of dyeing increases; Within 8～12 hours, playing stained positive region no longer changes.Obviously, very long dyeing time can not meet such as needs such as clinical operations and can to nerve fibre function, complete fast the requirements of one's work of quick identification.
Raman spectrum analysis method is based on Raman scattering effect, scattering spectrum analyze different from incident light frequency obtained to molecular vibration, rotation aspect information, and be applied to a kind of analytical approach that molecular structure is studied, can reflect content and chemical constitution and the variation of nucleic acid, protein, lipid in tiny organism sample.Yet, the raman spectral signal of the motor fiber of direct collection and sensory fibre is analyzed, do not find that between the Raman signal of two kinds of peripheral nerve fibers, there were significant differences, but simultaneously completely different between every curve of spectrum.According to the experience with Raman spectrum detection of biological sample and the result in many documents, the Raman signal of biological sample is very complicated, and raman scattering intensity formed by measured matter density in sweep limit, structure and the various factorss such as existence of impurity affect, even if cause, between the spectral signal of twice measurement of same position, also can there is certain little deviation.We also find by experiment, and the spectrum difference of the nerve section sample of difference in functionality itself may be equally very small, and the identification of being undertaken in histology by trickle signal difference is very difficult.
The essence of Karnovsky-Roots dyeing is by acetylthiocholine iodide and acetylcholinesterase (AChE) combination in fact, through the redox reaction with dyeing liquor, generating sepia copper ferrocyanide is deposited in the aixs cylinder of motor fibre, in the aixs cylinder of Sensory nerve fibre, owing to having AChE that chemical reaction can not occur, do not produce precipitation deposition, so make motion and Sensory nerve fibre can change and identify by spectral characteristic under light microscopic.
Motor fibre through Karnovsky-Roots dyeing all can produce Raman scattering signal in the peak position beyond the Raman scattering of itself, and along with the increase of dyeing time precipitates the increase of molecular amounts, raman scattering intensity is and increases progressively relation, can from complicated neural bio signal, show one's talent, avoid the interference of the signal difference of nerve fibre own.The feature of the Raman spectrum of nerve fibre is, at 550cm
-1, 1080cm
-1, 1110cm
-1, 1280cm
-1, 1440cm
-1, 1660cm
-1there is obvious raman scattering intensity, wherein with 1440cm
-1intensity maximum.Except the raman scattering intensity of nerve fibre itself, motor fiber, owing to containing AChE, after Karnovsky-Roots dyeing, produces copper ferrocyanide precipitation, therefore at 480cm in aixs cylinder
-1, 510cm
-1, 595cm
-1, 2110cm
-1, 2155cm
-1the raman scattering intensity from dyeing precipitation detected, wherein with 2100-2200cm
-1signal intensity maximum, with reference to the organism raman frequency table of comparisons, can determine 2100 cm
-1～2200 cm
-1for CN signal, the group reflecting conforms to the chemical composition of precipitate ferrous copper cyanider; In Sensory nerve fibre, owing to lacking AChE, copper ferrocyanide precipitation is gathered few, cannot observe the raman scattering intensity of corresponding peak position.The method of quick discriminating peripheral nerve character of the present invention completes based on aforementioned the principles of science.
The technical scheme that the present invention takes is: described method comprises the following steps: obtain perineural cross-section specimen, thickness is 1-2mm; Carry out frozen section, slice thickness is 20-30 μ m; Under micro-Raman spectroscopy, laser beam is focused on to nerve fibre section and carry out Raman spectrum collection; The Karnovsky-Roots 30min that dyes; Again under micro-Raman spectroscopy, by the laser beam of same wavelength, focus on nerve fibre section and carry out Raman spectrum collection; According to the character of the spectral results judgement nerve fibre before and after dyeing.
Described micro-Raman spectroscopy is HORIBA Jobin Yvon LabRam-1B type laser co-focusing micro-Raman spectroscopy.
Described micro-Raman spectroscopy is HORIBA Jobin Yvon XploRA-FDU type laser co-focusing micro-Raman spectroscopy.
Described laser beam wavelength is 532nm.
Described laser beam wavelength is 632.8nm.
Described Raman spectrum acquisition range is 1400-2200cm
-1, acquisition time is 10s.
Described Raman spectrum acquisition range is 1400-2200cm
-1, acquisition time is 50s.
The method of described judgement nerve fibre character is: to the Raman spectrum data collecting, use LabSpec software to carry out artificial baseline and process and computing machine polynomial iterative method smooth curve, obtain the final micro Raman spectra for data analysis, I after dyeing 30min
2100/ I
1440the nerve fibre of ratio >0.2 is motor fibre, otherwise is sensory fibre.
The method of described judgement nerve fibre character is: to the Raman spectrum data collecting, use LabSpec software to carry out artificial baseline and process and computing machine polynomial iterative method smooth curve, obtain the final micro Raman spectra for data analysis, after Karnovsky-Roots dyeing 30min, Raman spectrum is at 2100cm
-1-2160cm
-1occur that obvious raman scattering intensity is motor fibre, otherwise be Sensory nerve fibre.
The invention has the advantages that:
1, the method for discriminating peripheral nerve character of the present invention can be in shorter time identification peripheral nerve the character of motor fiber and sensory fibre, help scientific research and the various demands that need clinically two kinds of nerve fibres to differentiate.
2, the method for discriminating peripheral nerve character of the present invention has good sensitivity, specificity and accuracy.
Accompanying drawing explanation
Accompanying drawing 1 is that the embodiment of the present invention 2 is with I
2100/ I
1450differentiate the ROC curve of the diagnostic test of the front root of spinal cord or rear root.
Embodiment
Below in conjunction with embodiment, embodiment provided by the invention is elaborated.
A kind of method of utilizing Raman spectroscopy combined staining to differentiate fast peripheral nerve character of the present invention comprises the following steps:
1) obtain perineural cross-section specimen, thickness is 1-2mm;
2) carry out frozen section, slice thickness is 20-30 μ m;
3), under micro-Raman spectroscopy, laser beam is focused on to nerve fibre section and carry out Raman spectrum collection;
4) Karnovsky-Roots dyeing 30min, prescription of its dyeing liquor is as shown in table 1;
Table 1 prescription of its dyeing liquor
| Solution kind and concentration | Volume |
| 0.1 M acetylthiocholine iodide solution | 20 ml |
| 0.1 M pH6.5 phosphate buffer | 160 ml |
| 0.1 M sodium citrate solution | 10 ml |
| 30 mM copper-baths | 25 ml |
| 5 mM potassium ferricyanide solutions | 25 ml |
5), again under micro-Raman spectroscopy, by the laser beam of same wavelength, focus on nerve fibre section and carry out Raman spectrum collection;
6) according to the character of the spectral results judgement nerve fibre before and after dyeing, concrete criterion is as follows: after Karnovsky-Roots dyeing 30min, Raman spectrum is at 2100cm
-1-2160cm
-1occur that obvious raman scattering intensity is motor fibre, otherwise be Sensory nerve fibre; I after dyeing 30min
2100/ I
1440the nerve fibre of ratio >0.2 is motor fibre, otherwise is sensory fibre.Definition I
2100be 2100～2160cm
-1interior raman scattering intensity maximal value, I
1440for (1400 ± 15) cm
-1interior raman scattering intensity maximal value.
embodiment 1
experiment material
Get 20 of new zealand white rabbits, get respectively its bilateral sacrum 1 ventral root and rear each 20 routine samples.
experimental apparatus
HORIBA Jobin Yvon LabRam-1B type laser co-focusing micro-Raman spectroscopy (Institute of Analysis of Fudan University Raman laboratory is used in Raman spectrum collection; Basic parameter: excitation wavelength 632.8nm, power 4.3mw).
experimental technique
1) obtain perineural cross-section specimen, thickness is 1-2mm;
2) carry out frozen section, slice thickness is 30 μ m;
3) under micro-Raman spectroscopy, the laser beam that is 632.8nm by wavelength focuses on nerve fibre section and carries out Raman spectrum collection, and acquisition range is 1400-2200cm
-1, acquisition time is 50s;
4) Karnovsky-Roots dyeing 30min, prescription of its dyeing liquor is as shown in table 1;
5) again under micro-Raman spectroscopy, by the laser beam that wavelength is 632.8nm, focus on nerve fibre section and carry out Raman spectrum collection, acquisition range is 1400-2200cm
-1, acquisition time is 50s;
6) to the Raman spectrum data collecting, using LabSpec software to carry out artificial baseline processes and computing machine polynomial iterative method smooth curve, obtains the final micro Raman spectra for data analysis.According to the character of the spectral results judgement nerve fibre before and after dyeing, concrete criterion is as follows: after Karnovsky-Roots dyeing 30min, Raman spectrum is at 2100cm
-1-2160cm
-1occur that obvious raman scattering intensity is motor fibre, otherwise be Sensory nerve fibre; I after dyeing 30min
2100/ I
1440the nerve fibre of ratio >0.2 is motor fibre, otherwise is sensory fibre.
It should be noted that, in the present invention, first classification and numbering after drawing materials according to ventral root and rear Anatomy Properties, then select sample by the sampling of table of random number method before experiment and carry out frozen section, Raman spectrum collection, histochemical stain and statistical study.The check of neural coloration result completes after spectra collection, therefore can be because of the randomness of subjective selection bias impact sampling.
The method is to motorial sensitivity 100%, specificity 95%, accuracy 97.5%.
embodiment 2
experiment material
Get 20 of new zealand white rabbits, get respectively its bilateral sacrum 1 ventral root and rear each 20 routine samples.
experimental apparatus
HORIBA Jobin Yvon XploRA-FDU type laser co-focusing micro-Raman spectroscopy (Institute of Analysis of Fudan University Raman laboratory is used in Raman spectrum collection; Basic parameter: excitation wavelength 532nm, power 25mw).
experimental technique
1) obtain perineural cross-section specimen, thickness is 1-2mm;
2) carry out frozen section, slice thickness is 30 μ m;
3) under micro-Raman spectroscopy, the laser beam that is 532nm by wavelength focuses on nerve fibre section and carries out Raman spectrum collection, and acquisition range is 1400-2200cm
-1, acquisition time is 10s;
4) adopt Karnovsky-Roots dyeing 30min, prescription of its dyeing liquor is as shown in table 1;
5) again under micro-Raman spectroscopy, by the laser beam that wavelength is 532nm, focus on nerve fibre section and carry out Raman spectrum collection, acquisition range is 1400-2200cm
-1, acquisition time is 10s;
6) to the Raman spectrum data collecting, using LabSpec software to carry out artificial baseline processes and computing machine polynomial iterative method smooth curve, obtains the final micro Raman spectra for data analysis.According to the character of the spectral results judgement nerve fibre before and after dyeing, concrete criterion is as follows: after Karnovsky-Roots dyeing 30min, Raman spectrum is at 2100cm
-1-2160cm
-1occur that obvious raman scattering intensity is motor fibre, otherwise be Sensory nerve fibre; I after dyeing 30min
2100/ I
1440the nerve fibre of ratio >0.2 is motor fibre, otherwise is sensory fibre.
It should be noted that, in the present invention, first classification and numbering after drawing materials according to ventral root and rear Anatomy Properties, then select sample by the sampling of table of random number method before experiment and carry out frozen section, Raman spectrum collection, histochemical stain and statistical study.The check of neural coloration result completes after spectra collection, therefore can be because of the randomness of subjective selection bias impact sampling.
experimental result
Karnovsky-Roots dyeing gained Raman spectrum data I after 30 minutes in computed reliability demonstration test
2100/ I
1450ratio, and carry out average comparison, result
p<0.001, as shown in table 2.
30 minutes I of root Karnovsky-Roots dyeing after root and spinal cord before table 2 spinal cord
2100/ I
1450relatively
| ? | Root before spinal cord | Root after spinal cord | t | P |
| I 2100/I 1450 | 1.33±0.84 | 0.11±0.15 | 6.385 | <0.001 |
Anatomy Properties, as goldstandard, according to carrying out diagnostic test, is identified motor fibre with the method to each 20 number of cases of root after root before spinal cord and spinal cord.All correct identification of root before 20 spinal cords, after 20 spinal cords, root is correctly identified.Calculate sensitivity 100%, specificity 95%, accuracy 97.5%, as shown in table 3.The data that utilization is taken off after blind are drawn ROC curve, as shown in Figure 1, and I
2100/ I
1450=0.266 o'clock sensitivity=1,1-specificity=0.05; I
2100/ I
1450=0.411 o'clock sensitivity=0.95,1-specificity=0.05; I
2100/ I
1450=0.684 o'clock sensitivity=0.65,1-specificity=0.05; I
2100/ I
1450=0.725 o'clock sensitivity=0.65,1-specificity=0.
The diagnostic test that table 3 micro Raman spectra associating Karnovsky-Roots dyeing is differentiated neural character
| ? | Root before spinal cord | Root after spinal cord | Sum |
| Stained positive | 20 | 1 | 21 |
| Dyeing is negative | 0 | 19 | 19 |
| Sum | 20 | 20 | 40 |
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, do not departing under the prerequisite of the inventive method; can also make some improvement and supplement, these improvement and supplement and also should be considered as protection scope of the present invention.
Claims (9)
1. utilize Raman spectroscopy combined staining to differentiate fast a method for peripheral nerve character, it is characterized in that, described method comprises the following steps: obtain perineural cross-section specimen, thickness is 1-2mm; Carry out frozen section, slice thickness is 20-30 μ m; Under micro-Raman spectroscopy, laser beam is focused on to nerve fibre section and carry out Raman spectrum collection; The Karnovsky-Roots 30min that dyes; Again under micro-Raman spectroscopy, by the laser beam of same wavelength, focus on nerve fibre section and carry out Raman spectrum collection; According to the character of the spectral results judgement nerve fibre before and after dyeing.
2. method according to claim 1, is characterized in that, described micro-Raman spectroscopy is HORIBA Jobin Yvon LabRam-1B type laser co-focusing micro-Raman spectroscopy.
3. method according to claim 1, is characterized in that, described micro-Raman spectroscopy is HORIBA Jobin Yvon XploRA-FDU type laser co-focusing micro-Raman spectroscopy.
4. method according to claim 1, is characterized in that, described laser beam wavelength is 632.8nm.
5. method according to claim 1, is characterized in that, described laser beam wavelength is 532nm.
6. method according to claim 1, is characterized in that, described Raman spectrum acquisition range is 1400-2200cm
-1, acquisition time is 50s.
7. method according to claim 1, is characterized in that, described Raman spectrum acquisition range is 1400-2200cm
-1, acquisition time is 10s.
8. method according to claim 1, it is characterized in that, the method of described judgement nerve fibre character is: to the Raman spectrum data collecting, use LabSpec software to carry out artificial baseline and process and computing machine polynomial iterative method smooth curve, obtain the final micro Raman spectra for data analysis, I after dyeing 30min
2100/ I
1440the nerve fibre of ratio >0.2 is motor fibre, otherwise is sensory fibre.
9. method according to claim 1, it is characterized in that, the method of described judgement nerve fibre character is: to the Raman spectrum data collecting, use LabSpec software to carry out artificial baseline and process and computing machine polynomial iterative method smooth curve, obtain the final micro Raman spectra for data analysis, after Karnovsky-Roots dyeing 30min, Raman spectrum is at 2100cm
-1-2160cm
-1occur that obvious raman scattering intensity is motor fibre, otherwise be Sensory nerve fibre.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410405809.2A CN104142322A (en) | 2014-08-18 | 2014-08-18 | Method for quickly authenticating nature of peripheral nerve by raman spectra technique and dyeing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410405809.2A CN104142322A (en) | 2014-08-18 | 2014-08-18 | Method for quickly authenticating nature of peripheral nerve by raman spectra technique and dyeing |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104142322A true CN104142322A (en) | 2014-11-12 |
Family
ID=51851557
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410405809.2A Pending CN104142322A (en) | 2014-08-18 | 2014-08-18 | Method for quickly authenticating nature of peripheral nerve by raman spectra technique and dyeing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104142322A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116796159A (en) * | 2023-08-17 | 2023-09-22 | 浙江恒逸石化有限公司 | Dyeing effect prediction method, training method and device of dyeing effect prediction model |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1664564A (en) * | 2005-03-21 | 2005-09-07 | 江苏省人民医院 | Method for quick-speed in-situ authentication of peripheral nerve tracts |
| CN101246159A (en) * | 2008-03-07 | 2008-08-20 | 华中科技大学 | Method for fast identifying peripheral nerve bundle nature |
| CN102243154A (en) * | 2011-04-02 | 2011-11-16 | 中山大学附属第一医院 | Method for dyeing peripheral nerves slice |
-
2014
- 2014-08-18 CN CN201410405809.2A patent/CN104142322A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1664564A (en) * | 2005-03-21 | 2005-09-07 | 江苏省人民医院 | Method for quick-speed in-situ authentication of peripheral nerve tracts |
| CN101246159A (en) * | 2008-03-07 | 2008-08-20 | 华中科技大学 | Method for fast identifying peripheral nerve bundle nature |
| CN102243154A (en) * | 2011-04-02 | 2011-11-16 | 中山大学附属第一医院 | Method for dyeing peripheral nerves slice |
Non-Patent Citations (3)
| Title |
|---|
| 徐沁同等: "《周围神经显微结构的拉曼光谱和超光谱成像特征》", 《中国临床医学》 * |
| 徐沁同等: "《显微拉曼光谱技术联合Karnovsky-Roots染色法快速鉴别周围神经纤维的功能性质》", 《复旦学报（医学版）》 * |
| 王虎等: "《拉曼光谱法快速区分周围神经束性质》", 《中国误诊学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116796159A (en) * | 2023-08-17 | 2023-09-22 | 浙江恒逸石化有限公司 | Dyeing effect prediction method, training method and device of dyeing effect prediction model |
| CN116796159B (en) * | 2023-08-17 | 2023-11-17 | 浙江恒逸石化有限公司 | Dyeing effect prediction method, training method and device of dyeing effect prediction model |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sikirzhytski et al. | Multidimensional Raman spectroscopic signatures as a tool for forensic identification of body fluid traces: a review | |
| Kudryavtsev et al. | Combining MFD and PIE for accurate single‐pair Förster resonance energy transfer measurements | |
| AU2018231211A1 (en) | Analyte detection | |
| Xu et al. | Field-amplified sample stacking capillary electrophoresis with electrochemiluminescence applied to the determination of illicit drugs on banknotes | |
| CN109001180B (en) | Raman spectrum combined artificial intelligence high-throughput single cell analysis and identification method | |
| Lee et al. | Quantitative, label-free characterization of stem cell differentiation at the single-cell level by broadband coherent anti-Stokes Raman scattering microscopy | |
| CN100547383C (en) | A kind of laser double-mode micro-volume sample analyzing method and equipment therefor | |
| D’Arco et al. | Subcellular chemical and morphological analysis by stimulated Raman scattering microscopy and image analysis techniques | |
| CN104894222A (en) | Novel method for beacon-free detection of T4 PNKP (T4 polynucleotide kinase)/phosphatase and inhibitor of T4 PNKP/phosphatase on basis of fluorescent copper nanoparticles | |
| CN104914089A (en) | Method for realizing semi-quantitative analysis to trace mixture by use of surface enhanced raman spectroscopy | |
| Perroud et al. | Effect of bin time on the photon counting histogram for one‐photon excitation | |
| AU2019309923A1 (en) | Intramolecular kinetic probes | |
| CN101196464B (en) | Laser double-mode micro-volume sample analyzing method and its device | |
| Wu et al. | Recent advances in Raman spectroscopy for skin diagnosis | |
| George et al. | Raman spectroscopy for the determination of forensically important bio-fluids | |
| CN104142322A (en) | Method for quickly authenticating nature of peripheral nerve by raman spectra technique and dyeing | |
| CN103698310A (en) | Sperm acrosome zone Raman spectrum peak and use thereof | |
| Bénard et al. | Discrimination between healthy and tumor tissues on formalin-fixed paraffin-embedded breast cancer samples using IR imaging | |
| CN115718089A (en) | Method for rapidly identifying sample category based on flora Raman features | |
| Pansare et al. | Quality assessment of cryopreserved biospecimens reveals presence of intact biomolecules | |
| WO2021120518A1 (en) | Polymer material aging detection and process analysis method | |
| Drinovec et al. | Discrimination of marine algal taxonomic groups using delayed fluorescence spectroscopy | |
| CN100372947C (en) | Fluorescence correlation spectral detection method and device for transgenosis material | |
| CN107058082A (en) | A kind of single-molecule DNA Non-Destructive Testing chip based near field optics | |
| Tian et al. | Estimating time since deposition of bloodstains by scanning electrochemical microscopy |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20141112 |
|
| RJ01 | Rejection of invention patent application after publication |