CN201302549Y - Non-destructive detecting device for cell components and intercellular components - Google Patents
Non-destructive detecting device for cell components and intercellular components Download PDFInfo
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- CN201302549Y CN201302549Y CNU200920136665XU CN200920136665U CN201302549Y CN 201302549 Y CN201302549 Y CN 201302549Y CN U200920136665X U CNU200920136665X U CN U200920136665XU CN 200920136665 U CN200920136665 U CN 200920136665U CN 201302549 Y CN201302549 Y CN 201302549Y
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- elastic fibers
- extracellular matrix
- pair cell
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- harmonic signal
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- 210000003850 cellular structure Anatomy 0.000 title 1
- 210000004177 elastic tissue Anatomy 0.000 claims abstract description 19
- 238000001514 detection method Methods 0.000 claims abstract description 16
- 210000004027 cell Anatomy 0.000 claims description 27
- 230000005284 excitation Effects 0.000 claims description 14
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 claims description 11
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 claims description 11
- 210000002744 extracellular matrix Anatomy 0.000 claims description 11
- 238000012360 testing method Methods 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 241000931526 Acer campestre Species 0.000 claims description 2
- 230000003287 optical effect Effects 0.000 claims description 2
- 238000011897 real-time detection Methods 0.000 abstract description 2
- 102000008186 Collagen Human genes 0.000 abstract 2
- 108010035532 Collagen Proteins 0.000 abstract 2
- 229920001436 collagen Polymers 0.000 abstract 2
- 230000001066 destructive effect Effects 0.000 abstract 2
- 239000000835 fiber Substances 0.000 abstract 2
- 238000004043 dyeing Methods 0.000 description 3
- 238000007489 histopathology method Methods 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 230000004304 visual acuity Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 2
- 230000002950 deficient Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000003118 histopathologic effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
本实用新型涉及一种对细胞和细胞外间质成分进行同时无损探测的装置，其特征在于：激光器发出的光通过一个双色分光镜入射到扫描器的扫描镜组上，通过扫描镜组的光束由物镜聚焦到检测样品上；激光与检测样品产生的双光子激发荧光和二次谐波信号反向通过相同的物镜收集，再由前述扫描镜组及双色分光镜入射到一个半透半反镜上，将双光子激发荧光和二次谐波信号组成的发射信号光分成两路，这两路通过滤波片和光电倍增管转换成电信号并接至计算机的输入端，最后由计算机同时显示细胞、弹力纤维和胶原纤维的微结构。该装置能够在分子水平上对细胞、胶原纤维和弹力纤维的显微结构进行高分辨率、高对比度、无损、快速和实时的检测。
The utility model relates to a device for simultaneous non-destructive detection of cells and extracellular interstitial components. Focused on the detection sample by the objective lens; the two-photon excited fluorescence and the second harmonic signal generated by the laser and the detection sample are collected through the same objective lens in reverse, and then incident on a half-transparent mirror by the aforementioned scanning mirror group and dichroic beam splitter Above, the emitted signal light composed of two-photon excited fluorescence and second harmonic signal is divided into two paths, which are converted into electrical signals through filters and photomultiplier tubes and connected to the input terminal of the computer, and finally the computer simultaneously displays the cells , elastic fibers and collagen fibers microstructure. The device can perform high-resolution, high-contrast, non-destructive, rapid and real-time detection of the microstructure of cells, collagen fibers and elastic fibers at the molecular level.
Description
Technical field
The utility model relates to a kind of pair cell and the extracellular matrix composition carries out the device of lossless detection simultaneously.
Technical background
Present clinical medical pathology department every day all can pair cell, the microstructure of extracellular matrix principal ingredient collagenous fibres and elastic fibers is observed and detected.The Histopathology method that clinical medical detection method is normally traditional.The histopathologic slide that is used for liver histopathological analysis need obtain through a series of processes such as fixing, paraffin embedding, dewaxing, dyeing, these processes have not only been destroyed some relevant useful material true forms of tissue, and the time that needs is often long, each step institute's time spent all be with hour, or even calculate with the sky.In addition, sample disposal is incorrect and dyeing time is not sure all might cause check result to lose accuracy.If think to survey simultaneously the microstructure of cell, collagenous fibres and elastic fibers, needs are observed after using diverse dyestuff that histotomy is dyeed respectively again.Overcome the Histopathology method defective, seek a kind of high resolving power, microstructural new device harmless and that detect cell, collagenous fibres and elastic fibers in real time is target and the dream that a large amount of researchers and clinician make joint efforts always.
Summary of the invention
The purpose of this utility model provides a kind of pair cell and the extracellular matrix composition carries out the device of lossless detection simultaneously, this device adopts two-photon fluorescence excitation that light and matter interaction launch and second harmonic signal and pair cell and extracellular matrix composition microstructure are carried out the while lossless detection, can be on molecular level the microstructure of pair cell, collagenous fibres and elastic fibers carry out high resolving power, high-contrast, harmless, quick and real-time detection.
The technical solution of the utility model is achieved in that the light that laser instrument sends incides by a double-colored spectroscope on the scanning mirror group of scanner, the light beam by the scanning mirror group by object lens focusing on test sample; Laser is oppositely collected by identical object lens with two-photon fluorescence excitation and second harmonic signal that test sample produces, incide on the semi-transparent semi-reflecting lens by aforementioned scanning mirror group and double-colored spectroscope again, the light that transmits of two-photon fluorescence excitation and second harmonic signal composition is divided into two-way, leading up to filter plate 1 and photomultiplier 1 converts the second harmonic signal of collagenous fibres to electric signal A, leading up to filter plate 2 and photomultiplier in addition becomes electric signal B with the two-photon fluorescence excitation conversion of signals of cell and elastic fibers, described electric signal A and electric signal B are connected to input end and computer respectively, at last by computing machine while showed cell, the microstructure of elastic fibers and collagenous fibres.
Remarkable advantage of the present utility model is: its microstructure of input of utilizing the endogenous molecule interaction generation of laser and cell and extracellular matrix composition, therefore, do not need processing such as dyeing, directly can realize the high resolving power to sample, harmless and detection in real time.Though the microstructural detection of cell and elastic fibers all is by the two-photon fluorescence excitation signal, but the structure of the two has tangible difference, with the human body is example, the minimum diameter of human body cell is approximately 4 μ m, the similar sphere of shape, and under the Electronic Speculum, the diameter of elastic fibers has only 1-3nm, become wavy, therefore, the microstructure of the two can obviously distinguish, if select suitable optical maser wavelength, make collagenous fibres based on the emission second harmonic signal, can obtain the high-contrast microstructure of cell, collagenous fibres and elastic fibers.In addition, the use of scanner makes device have the function of fast detecting.
Description of drawings
Fig. 1 is the organigram of the utility model embodiment.
Embodiment
Below in conjunction with accompanying drawing embodiment of the present utility model is further elaborated, so that the utility model becomes apparent.
As shown in Figure 1, the light that laser instrument sends incides by a double-colored spectroscope on the scanning mirror group of scanner, the light beam by the scanning mirror group by object lens focusing on test sample; Laser is oppositely collected by identical object lens with two-photon fluorescence excitation and second harmonic signal that test sample produces, incide on the semi-transparent semi-reflecting lens by aforementioned scanning mirror group and double-colored spectroscope again, the light that transmits of two-photon fluorescence excitation and second harmonic signal composition is divided into two-way, leading up to filter plate 1 and photomultiplier 1 converts the second harmonic signal of collagenous fibres to electric signal A, leading up to filter plate 2 and photomultiplier in addition becomes electric signal B with the two-photon fluorescence excitation conversion of signals of cell and elastic fibers, described electric signal A and electric signal B are connected to input end and computer respectively, at last by computing machine while showed cell, the microstructure of elastic fibers and collagenous fibres.
In most preferred embodiment of the present utility model, the wavelength of laser instrument should be selected 810nm for use, this moment collagenous fibres based on the emission second harmonic signal, avoid pair cell particularly to the detection of elastic fibers microstructure high-contrast.If the wavelength of laser instrument is selected 810nm for use, double-colored spectroscope need see through the incident light greater than 700nm, reflects the emission light less than 700nm simultaneously.The filter range of filter plate 1 is realized the collection to the collagenous fibres second harmonic signal between 400-410nm, and the filter range of filter plate 2 is realized the collection of pair cell and elastic fibers two-photon fluorescence excitation signal between 440-650nm.
As long as above-mentioned laser instrument satisfies the condition of light and cell, collagenous fibres and elastic fibers interaction generation two-photon fluorescence excitation and second harmonic signal, can be dissimilar laser instruments, as different types of laser instruments such as femtosecond laser instrument, picosecond lasers.
Be provided with light intensity control and total reflective mirror between above-mentioned laser instrument and double-colored spectroscope, above-mentioned light intensity control is made up of 1/2nd wave plates and Glan prism.
The utility model is reasonable in design, is skillfully constructed, and has vast potential for future development and bigger dissemination.
Claims (5)
1. pair cell and extracellular matrix composition carry out the device of lossless detection simultaneously, it is characterized in that: the light that laser instrument sends incides by a double-colored spectroscope on the scanning mirror group of scanner, the light beam by the scanning mirror group by object lens focusing on test sample; Laser is oppositely collected by identical object lens with two-photon fluorescence excitation and second harmonic signal that test sample produces, incide on the semi-transparent semi-reflecting lens by aforementioned scanning mirror group and double-colored spectroscope again, the light that transmits of two-photon fluorescence excitation and second harmonic signal composition is divided into two-way, leading up to filter plate 1 and photomultiplier 1 converts the second harmonic signal of collagenous fibres to electric signal A, leading up to filter plate 2 and photomultiplier in addition becomes electric signal B with the two-photon fluorescence excitation conversion of signals of cell and elastic fibers, described electric signal A and electric signal B are connected to input end and computer respectively, at last by computing machine while showed cell, the microstructure of elastic fibers and collagenous fibres.
2. pair cell according to claim 1 and extracellular matrix composition carry out the device of lossless detection simultaneously, it is characterized in that: the optical maser wavelength that described laser instrument sends equals 810nm, described double-colored spectroscope can see through the incident light greater than 700nm, reflect emission light simultaneously less than 700nm, the filter range of filter plate 1 is between 400-410nm, realization is to the collection of collagenous fibres second harmonic signal, and the filter range of filter plate 2 is realized the collection of pair cell and elastic fibers two-photon fluorescence excitation signal between 440-650nm.
3. pair cell according to claim 1 and extracellular matrix composition carry out the device of lossless detection simultaneously, and it is characterized in that: described laser instrument is femto-second laser or picosecond laser.
4. pair cell according to claim 1 and extracellular matrix composition carry out the device of lossless detection simultaneously, it is characterized in that: be provided with light intensity control and total reflective mirror between described laser instrument and double-colored spectroscope.
5. pair cell according to claim 4 and extracellular matrix composition carry out the device of lossless detection simultaneously, it is characterized in that described light intensity control is made up of 1/2nd wave plates and Glan prism.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU200920136665XU CN201302549Y (en) | 2009-02-12 | 2009-02-12 | Non-destructive detecting device for cell components and intercellular components |
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| CNU200920136665XU CN201302549Y (en) | 2009-02-12 | 2009-02-12 | Non-destructive detecting device for cell components and intercellular components |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102297854A (en) * | 2011-05-23 | 2011-12-28 | 公安部第一研究所 | High-efficiency multi-mode laser-induced fluorescence optical path exciting system |
| CN103616359A (en) * | 2013-12-04 | 2014-03-05 | 中国检验检疫科学研究院 | Fluorescent quantitative detection method for green fluorescent protein (GFP) |
| CN104596964A (en) * | 2015-02-03 | 2015-05-06 | 黑龙江大学 | Device and method for measuring transient absorbency of semitransparent thin film material |
| CN104758058A (en) * | 2015-03-11 | 2015-07-08 | 苏州大学 | Synchronous pulse laser microimaging observation device for mechanical stress deformation of blood cells |
| CN105738254A (en) * | 2016-02-03 | 2016-07-06 | 苏州大学 | Mechanobiological coupling testing system and method |
| CN106841145A (en) * | 2017-02-28 | 2017-06-13 | 中国中医科学院医学实验中心 | Second Harmonic Imaging combined with fluorescent is imaged on the application in positioning material Transdermal absorption |
| CN106990077A (en) * | 2017-06-15 | 2017-07-28 | 福建师范大学 | It is a kind of based on the micro- high throughput screening system with microfluidic chip technology of multi-photon |
| CN109073539A (en) * | 2016-05-03 | 2018-12-21 | 文塔纳医疗系统公司 | System and method for monitoring reagent concentration |
| CN109813654A (en) * | 2019-02-20 | 2019-05-28 | 北京工业大学 | The time discrimination measurement device of material surface state second_harmonic generation |
| CN110082330A (en) * | 2019-05-16 | 2019-08-02 | 福建师范大学 | Combined system is imaged in a kind of laser optical tweezer Raman spectrum and multi-photon |
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2009
- 2009-02-12 CN CNU200920136665XU patent/CN201302549Y/en not_active Expired - Fee Related
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102297854A (en) * | 2011-05-23 | 2011-12-28 | 公安部第一研究所 | High-efficiency multi-mode laser-induced fluorescence optical path exciting system |
| CN103616359A (en) * | 2013-12-04 | 2014-03-05 | 中国检验检疫科学研究院 | Fluorescent quantitative detection method for green fluorescent protein (GFP) |
| CN104596964A (en) * | 2015-02-03 | 2015-05-06 | 黑龙江大学 | Device and method for measuring transient absorbency of semitransparent thin film material |
| CN104596964B (en) * | 2015-02-03 | 2017-05-31 | 黑龙江大学 | A kind of translucent thin membrane material transient absorbance degree measurement apparatus and method |
| CN104758058A (en) * | 2015-03-11 | 2015-07-08 | 苏州大学 | Synchronous pulse laser microimaging observation device for mechanical stress deformation of blood cells |
| CN105738254A (en) * | 2016-02-03 | 2016-07-06 | 苏州大学 | Mechanobiological coupling testing system and method |
| CN105738254B (en) * | 2016-02-03 | 2019-07-12 | 苏州大学 | A kind of Mechanobiology coupling testing system and method |
| CN109073539A (en) * | 2016-05-03 | 2018-12-21 | 文塔纳医疗系统公司 | System and method for monitoring reagent concentration |
| CN106841145A (en) * | 2017-02-28 | 2017-06-13 | 中国中医科学院医学实验中心 | Second Harmonic Imaging combined with fluorescent is imaged on the application in positioning material Transdermal absorption |
| CN106841145B (en) * | 2017-02-28 | 2020-06-12 | 中国中医科学院医学实验中心 | Application of second harmonic imaging and fluorescence imaging in positioning substance transdermal absorption |
| CN106990077A (en) * | 2017-06-15 | 2017-07-28 | 福建师范大学 | It is a kind of based on the micro- high throughput screening system with microfluidic chip technology of multi-photon |
| CN109813654A (en) * | 2019-02-20 | 2019-05-28 | 北京工业大学 | The time discrimination measurement device of material surface state second_harmonic generation |
| CN109813654B (en) * | 2019-02-20 | 2021-10-01 | 北京工业大学 | Time-resolved measurement device for second harmonic generation of material surface states |
| CN110082330A (en) * | 2019-05-16 | 2019-08-02 | 福建师范大学 | Combined system is imaged in a kind of laser optical tweezer Raman spectrum and multi-photon |
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Granted publication date: 20090902 Termination date: 20120212 |