WO2020104975A4 - Method and system for analysis of organic material by auto-cathodoluminescence - Google Patents
Method and system for analysis of organic material by auto-cathodoluminescence Download PDFInfo
- Publication number
- WO2020104975A4 WO2020104975A4 PCT/IB2019/060004 IB2019060004W WO2020104975A4 WO 2020104975 A4 WO2020104975 A4 WO 2020104975A4 IB 2019060004 W IB2019060004 W IB 2019060004W WO 2020104975 A4 WO2020104975 A4 WO 2020104975A4
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cathodoluminescence
- parameters
- function
- specimen
- generating
- Prior art date
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/225—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion
- G01N23/2251—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion using incident electron beams, e.g. scanning electron microscopy [SEM]
- G01N23/2254—Measuring cathodoluminescence
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Analysing Materials By The Use Of Radiation (AREA)
Abstract
Analyzing isolated collagen gel samples of a rat-tail and bovine hide, we demonstrated in situ detection of spectrally deconvoluted auto-cathodoluminescence signatures of specific molecular content with precise spatial localization over a maximum field of view of 300 µm. Correlation of the secondary electron and the hyperspectral images proved ~40 nm resolution in the optical channel, obtained due to a short carrier diffusion length, suppressed by fibril dimensions and poor electrical conductivity specific to their organic composition. By correlating statistically analyzed auto-cathodoluminescence with a mass spectroscopy data, we differentiated spectral signatures of two extracellular matrix networks - human fibrin complex and rat tail collagen isolate, and uncovered differences in protein distributions of isolated extracellular matrix networks of heterogeneous populations. Furthermore, we demonstrated how cathodoluminescence can monitor progress of a cell-remodeling process from a rat collagenous into human collagenous gel, revealing changes in complex heterogeneous biological composition confirmed by mass spectroscopy.
Claims
1. A method of characterizing the molecular structure of a label-free biological specimen comprising a plurality of molecules, the method comprising: scanning an electron beam probe on to the specimen to thereby excite a portion of said plurality of molecules having no labeling applied thereto, the portion defining an interaction volume; collecting cathodoluminescence light emitted by said interaction volume upon the electron irradiation; spectrally resolving said cathodoluminescence to generate cathodoluminescence spectra; and, generating a hyperspectral cathodoluminescence image from the cathodoluminescence spectra wherein spectral information is added as a third dimension of values to a two- dimensional spatial image.
2. The method according to claim 1, further comprising: selecting a mathematical fitting function of the cathodoluminescence wavelength and M parameters, where M is larger or equal to 1 ; fitting the cathodoluminescence spectra with said function of M parameters, so that the mathematical function approximates the spectral shape as a function of the said M parameters; deriving a statistical distribution of one or more of the M parameters so that the statistical distribution of the said one or more M parameters characterizes a fingerprint of said specimen.
3. The method of claim 2, further comprising selecting one of the M parameters and generating a spatially resolved image such that each pixel has the value of the selected one of the M parameters.
4. The method of claims 2 or 3, wherein the selected one of M parameters is one of: wavelength, frequency, or energy.
2
5. The method of any of claims 2-4, further comprising overlaying spatially resolved images of at least two different M parameters.
6. The method of claim 2, further comprising generating a histogram of the selected one of the M parameters for a spatially resolved image of the selected one of the M parameters.
7. The method according to one of the preceding claims, wherein said mathematical function is a normal distribution (Gaussian function), a Lorentzian function, or a pseudo- Voigt function.
8. The method according to one of the preceding claims, further comprising a step of correlating the statistical distribution to a second statistical distribution obtained on a second specimen, different from the said specimen.
9. The method according to one of the preceding claims, further comprising: separating the cathodoluminescence spectra into defined frequency bands thereby generating hyperspectral CL data components; and, generating spatially resolved images form the hyperspectral CL data components.
10. The method according to claim 9, further comprising overlying spatially resolved images of at least two different hyperspectral CL data components.
11. The method according to claim 9, further comprising generating a plot of count number versus wavelength for a plurality of different hyperspectral CL data components.
12. The method according to claim 11, further comprising fitting the plot to a mathematical function.
13. The method according to claim 12, further comprising defining a fingerprint of the specimen from the deviation between the plot to the mathematical function.
14. The method according to claim 13, further comprising performing mass
spectroscopy analysis of the specimen.
3
15. A method for characterizing biological sample, comprising: irradiating the sample with a scanning electron beam, thereby causing the sample to emit cathodoluminescence photons; collecting the cathodoluminescence photons and forming a light beam therefrom; separating the light beam into light spectra; sensing the light spectra to generate a plurality of signals, each corresponding to a different band of the light spectra; overlaying at least a subset of the plurality of signals to generate a spatial distribution image.
16. The method of claim 15, further comprising measuring a histogram of the spatial distribution image.
17. The method of claim 15, further comprising fitting the histogram to a mathematical function.
18. The method of claim 17, wherein the mathematical function comprises one of: a normal distribution (Gaussian function), a Lorentzian function, or a pseudo- Voigt function.
19. The method of claim 17, further comprising generating a sample signature from the differences between the histogram and the mathematical function.
20. The method of claim 19, wherein the differences comprise central position and statistical distribution of width of the histogram.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB1818914.2 | 2018-11-20 | ||
| GBGB1818914.2A GB201818914D0 (en) | 2018-11-20 | 2018-11-20 | Apparatus and methods for high-resolution analysis of organic material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO2020104975A1 WO2020104975A1 (en) | 2020-05-28 |
| WO2020104975A4 true WO2020104975A4 (en) | 2020-07-16 |
Family
ID=64739954
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/IB2019/060004 WO2020104975A1 (en) | 2018-11-20 | 2019-11-20 | Method and system for analysis of organic material by auto-cathodoluminescence |
Country Status (2)
| Country | Link |
|---|---|
| GB (1) | GB201818914D0 (en) |
| WO (1) | WO2020104975A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP7444481B2 (en) * | 2019-05-09 | 2024-03-06 | アットライト エージー | cathodoluminescence electron microscope |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4929041A (en) * | 1989-01-09 | 1990-05-29 | Johnston Pump/General Valve, Inc. | Cathodoluminescence system for use in a scanning electron microscope including means for controlling optical fiber aperture |
| CN103261879B (en) * | 2010-10-01 | 2015-04-22 | 安托莱特公司 | Deconvolution of time-gated cathodoluminescence images |
| US10018579B1 (en) * | 2015-02-18 | 2018-07-10 | Kla-Tencor Corporation | System and method for cathodoluminescence-based semiconductor wafer defect inspection |
-
2018
- 2018-11-20 GB GBGB1818914.2A patent/GB201818914D0/en not_active Ceased
-
2019
- 2019-11-20 WO PCT/IB2019/060004 patent/WO2020104975A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| GB201818914D0 (en) | 2019-01-02 |
| WO2020104975A1 (en) | 2020-05-28 |
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