CN102621117A - Living cell laser scanning co-focusing microscope imaging system - Google Patents
Living cell laser scanning co-focusing microscope imaging system Download PDFInfo
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- CN102621117A CN102621117A CN201210061262XA CN201210061262A CN102621117A CN 102621117 A CN102621117 A CN 102621117A CN 201210061262X A CN201210061262X A CN 201210061262XA CN 201210061262 A CN201210061262 A CN 201210061262A CN 102621117 A CN102621117 A CN 102621117A
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Abstract
The invention relates to a living cell laser scanning co-focusing microscope imaging system, which is characterized by comprising a laser scanning co-focusing microscope and a fluorescent signal collecting device, wherein the fluorescent signal collecting device comprises a culture dish, a converging lens and a reflecting type narrow-band light filter, the converging lens and the reflecting type narrow-band light filter are arranged in the culture dish, the bottom of the culture dish is provided with a culture dish bottom plate used for placing cells to be tested, the converging lens is fixedly arranged on the culture dish through a lens clamping frame, the cells to be tested on the culture dish bottom plate are positioned on the focal plane of the converging lens, the reflecting type narrow-band light filter is positioned right above the converging lens, and an objective lens of the laser scanning co-focusing microscope is positioned right under the culture dish bottom plate. The living cell laser scanning co-focusing microscope imaging system has the advantages that the structure is compact, the precision is high, the fluorescent signal collection efficiency of the laser scanning co-focusing microscope can be effectively improved, the phototoxicity and the light bleaching degree are reduced, and a technical means is provided for obtaining reliable experiment results.
Description
Technical field
The present invention relates to a kind of living cells laser scanning co-focusing micro imaging system.
Background technology
The living cells optical image technology is look unfamiliar a kind of important means of reason, pathomechanism of research cellular layer.In the living cells optical imagery, phototoxicity and photobleaching are two important factors, and the accuracy of experimental result is played crucial effects, and be especially all the more so to long-time observation of cell physiology, pathological state.Phototoxicity can influence cell activity, and experimental result is produced indirect influence, and photobleaching can influence the fluorescence signal that obtains, and experimental result is produced directly influence.Therefore, reduce phototoxicity and photobleaching degree as far as possible become the common problem of paying close attention to of people in the living cells optical image technology.Though can alleviate phototoxicity and photobleaching degree through reducing the laser irradiation light dosage, meanwhile also can reduce the signal to noise ratio (S/N ratio) of signal.So the collecting efficiency that improves fluorescence signal is to improve signal to noise ratio (S/N ratio) and the important channel that reduces phototoxicity and photobleaching degree.
Summary of the invention
Deficiency in view of prior art the object of the present invention is to provide a kind of living cells laser scanning co-focusing micro imaging system with high collection efficiency.
To achieve these goals; Technical scheme of the present invention is: a kind of living cells laser scanning co-focusing micro imaging system; Comprise laser scanning co-focusing microscope and fluorescence signal harvester, said fluorescence signal harvester comprises double dish and is arranged at convergent lens and the reflective narrow band pass filter in the double dish; The bottom of said double dish is provided with the double dish base plate that is used to place cell to be measured; Said convergent lens is fixed on the double dish through the lens holding frame, and the cell to be measured on the said double dish base plate is positioned on the focal plane of convergent lens; Said reflective narrow band pass filter be positioned at convergent lens directly over; The object lens of said laser scanning co-focusing microscope be positioned at the double dish base plate under.
Further, said reflective narrow band pass filter is arranged on the lens holding frame through micro electronmechanical angular adjustment ring, and said lens holding frame is provided with the mems chip interface.
Further, said double dish bottom plate glue sticks on the double dish.
Further, said double dish is provided with the nutrient solution inflow interface and the nutrient solution outflow interface that is used to export nutrient solution that is used to import nutrient solution.
Further, said laser scanning co-focusing microscope comprises laser instrument, collimation lens, exciter filter, dichroic mirror, object lens, convergent lens, pin hole and photomultiplier.
The present invention has the following advantages: this living cells laser scanning co-focusing micro imaging system compact conformation, precision height; Can effectively improve laser scanning co-focusing microscope the fluorescence signal collecting efficiency, reduced phototoxicity and photobleaching degree, for obtaining reliable experimental result a kind of technological means is provided.
Description of drawings
Fig. 1 is the organigram of the embodiment of the invention.
Among the figure: 1-lens holding frame, the reflective narrow band pass filter of 2-, the micro electronmechanical angular adjustment ring of 3-, 4-mems chip interface, 5-convergent lens; The 6-nutrient solution flows into interface, 7-cell to be measured, 8-object lens, 9-dichroic mirror, 10-convergent lens; The 11-pin hole, 12-photomultiplier, 13-exciter filter, 14-collimation lens, 15-laser instrument; 16-forward scattering fluorescence, 17-double dish base plate, the 18-nutrient solution flows out interface, 19-double dish.
Embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is done further elaboration.
With reference to figure 1; A kind of living cells laser scanning co-focusing micro imaging system; Comprise laser scanning co-focusing microscope and fluorescence signal harvester, said fluorescence signal harvester comprises double dish 19 and is arranged at the convergent lens 5 and reflective narrow band pass filter 2 in the double dish 19; The bottom of said double dish 19 is provided with the double dish base plate 17 that is used to place cell 7 to be measured; Said convergent lens 5 is fixed on the double dish 19 through lens holding frame 1, and the cell to be measured 7 on the said double dish base plate 17 is positioned on the focal plane of convergent lens 5; Said reflective narrow band pass filter 2 be positioned at convergent lens 5 directly over; The object lens 8 of said laser scanning co-focusing microscope be positioned at double dish base plate 17 under.
In the present embodiment, said reflective narrow band pass filter 2 is arranged on the lens holding frame 1 through micro electronmechanical angular adjustment ring 3, and said lens holding frame 1 is provided with mems chip interface 4.Said reflective narrow band pass filter 2 is a kind of to the long-wave band transmission, to the optical filter of short-wave band reflection; The MEMS that said micro electronmechanical angular adjustment ring 3 is a kind of adjusting angles, the experimenter controls micro electronmechanical angular adjustment ring 3 through mems chip interface 4.In order to satisfy the requirement of the short operating distance of high power objective 8, said double dish base plate 17 adopts the viscose glue modes to be fixed on the double dish 19 of cell.For can be in the longer time physiology, the pathological characters of observation of cell; Said double dish 19 is provided with the nutrient solution inflow interface 6 and the nutrient solution outflow interface 18 that is used to export nutrient solution that is used to import nutrient solution, and nutrient solution flows out from nutrient solution inflow interface 6 entering double dish 19 and from nutrient solution outflow interface 18.
Laser scanning co-focusing microscope (LSCM) is on fluorescent microscope imaging basis, to have installed laser scanning device additional, uses ultraviolet or excited by visible light fluorescence probe to obtain the fluoroscopic image of the inner microtexture of cell or tissue.The basic composition of laser scanning co-focusing microscope: optical microscope part, laser instrument, scanister, signal supervisory instrument and computer system.In the present embodiment, said laser scanning co-focusing microscope comprises laser instrument 15, collimation lens 14, exciter filter 13, dichroic mirror 9, object lens 8, convergent lens 10, pin hole 11 and photomultiplier 12 etc.
The principle of work of present embodiment: when exciting light through collimation pass through 14, exciter filter 13, dichroic mirror 9 and object lens 8 converge to 7 last times of cell to be measured, the fluorescence probe that loads in the activated cell produces forward scattering fluorescence 16 and back scattering fluorescence; Forward scattering fluorescence 16 becomes directional light after acting on through the convergent lens in the double dish 19 5 with exciting light; Controlling micro electronmechanical angular adjustment ring 3 makes reflective narrow band pass filter 2 perpendicular to directional light; Since the 16 high reflections of 2 pairs of forward scattering fluorescence of reflective narrow band pass filter, and to the high transmission of exciting light, so forward scattering fluorescence 16 is reflected back toward the convergent lens 5 in the double dish 19, exciting light then scatters to external environment; Again focus on shot point under the effect of the convergent lens 5 of the forward scattering fluorescence 16 that is reflected in double dish 19, and with back scattering fluorescence directive object lens 8 together; Focus on and surveyed by the receiving plane of photomultiplier 12 through dichroic mirror 9 and convergent lens 10 backs through pin hole 11 backs.
Living cells laser scanning co-focusing micro imaging system compact conformation of the present invention, precision height; It has made up a two confocal arrangement through reflective narrow band pass filter in the double dish 19 2 and convergent lens 5; Can collect forward scattering and backward scattered fluorescence simultaneously; So can improve collection efficiency effectively, reduce phototoxicity and photobleaching degree, a kind of technological means is provided for obtaining reliable experimental result.
The above is merely preferred embodiment of the present invention, and all equalizations of doing according to claim of the present invention change and modify, and all should belong to covering scope of the present invention.
Claims (5)
1. living cells laser scanning co-focusing micro imaging system; It is characterized in that: comprise laser scanning co-focusing microscope and fluorescence signal harvester, said fluorescence signal harvester comprises double dish and is arranged at convergent lens and the reflective narrow band pass filter in the double dish; The bottom of said double dish is provided with the double dish base plate that is used to place cell to be measured; Said convergent lens is fixed on the double dish through the lens holding frame, and the cell to be measured on the said double dish base plate is positioned on the focal plane of convergent lens; Said reflective narrow band pass filter be positioned at convergent lens directly over; The object lens of said laser scanning co-focusing microscope be positioned at the double dish base plate under.
2. a kind of living cells laser scanning co-focusing micro imaging system according to claim 1; It is characterized in that: said reflective narrow band pass filter is arranged on the lens holding frame through micro electronmechanical angular adjustment ring, and said lens holding frame is provided with the mems chip interface.
3. a kind of living cells laser scanning co-focusing micro imaging system according to claim 1 and 2, it is characterized in that: said double dish bottom plate glue sticks on the double dish.
4. a kind of living cells laser scanning co-focusing micro imaging system according to claim 1 is characterized in that: said double dish is provided with the nutrient solution inflow interface and the nutrient solution outflow interface that is used to export nutrient solution that is used to import nutrient solution.
5. a kind of living cells laser scanning co-focusing micro imaging system according to claim 1, it is characterized in that: said laser scanning co-focusing microscope comprises laser instrument, collimation lens, exciter filter, dichroic mirror, object lens, convergent lens, pin hole and photomultiplier.
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| CN201210061262.XA CN102621117B (en) | 2012-03-09 | 2012-03-09 | Living cell laser scanning co-focusing microscope imaging system |
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| CN201210061262.XA CN102621117B (en) | 2012-03-09 | 2012-03-09 | Living cell laser scanning co-focusing microscope imaging system |
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| CN102621117B CN102621117B (en) | 2014-03-12 |
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102944540A (en) * | 2012-10-11 | 2013-02-27 | 中国科学院西安光学精密机械研究所 | Three-dimensional imaging system and method in deep scattering medium |
| CN103033920A (en) * | 2012-12-03 | 2013-04-10 | 苏州大猫单分子仪器研发有限公司 | Far infrared temperature jump microscope |
| CN104178420A (en) * | 2014-09-05 | 2014-12-03 | 中国科学技术大学 | Living cell culture environment control system for microscope |
| CN104181089A (en) * | 2013-05-22 | 2014-12-03 | 中国石油化工股份有限公司 | Equipment for scanning facial porosity of rock and method thereof |
| CN104535547A (en) * | 2014-11-29 | 2015-04-22 | 北京工业大学 | Confocal optical path portable apparatus for observing micro-fluidic chip |
| CN106568755A (en) * | 2016-11-06 | 2017-04-19 | 浙江大学 | Near infrared laser scanning confocal microscopic imaging system |
| CN107683430A (en) * | 2015-06-02 | 2018-02-09 | 生命技术公司 | System and method for generating structureization illumination image |
| CN107688014A (en) * | 2016-09-21 | 2018-02-13 | 北京大学 | A kind of cell imaging method |
| CN107702661A (en) * | 2017-09-19 | 2018-02-16 | 福建师范大学 | A kind of white light self-interference Surface testing instrument |
| CN109355177A (en) * | 2018-11-29 | 2019-02-19 | 苏州度奕星科技有限公司 | A kind of living cells monitoring device |
| CN111065950A (en) * | 2017-09-15 | 2020-04-24 | 敏捷焦点设计有限责任公司 | Dynamic focus and zoom system for wide-field, confocal and multiphoton microscopes |
| CN111258044A (en) * | 2020-02-18 | 2020-06-09 | 清华大学 | Mirror scanning light field-based 4 pi-like microscopic imaging method |
| CN112268505A (en) * | 2020-10-22 | 2021-01-26 | 江苏集萃微纳自动化系统与装备技术研究所有限公司 | Microscopic operation system based on full-field optical coherence tomography technology |
| WO2021120509A1 (en) * | 2019-12-18 | 2021-06-24 | 江南大学 | Instrument and method for counting microbial cells |
| US11346763B2 (en) | 2019-12-18 | 2022-05-31 | Jiangnan University | Apparatus and method for microbial cell counting |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0538551A2 (en) * | 1991-10-24 | 1993-04-28 | Toa Medical Electronics Co., Ltd. | Flow imaging cytometer |
| WO1994029695A1 (en) * | 1993-06-08 | 1994-12-22 | Gjelsnes Oddbjoern | Liquid flow cytometer |
| WO1999037999A1 (en) * | 1998-01-27 | 1999-07-29 | Wisconsin Alumni Research Foundation | Signal enhancement for fluorescence microscopy |
| JP2000227556A (en) * | 1999-02-05 | 2000-08-15 | Japan Science & Technology Corp | Microscope |
| CN1605856A (en) * | 2004-11-18 | 2005-04-13 | 上海交通大学 | Laser fluorescence correlation spectrum unimolecular analyzer |
| JP2006011045A (en) * | 2004-06-25 | 2006-01-12 | Olympus Corp | Total reflection microscope |
| CN1798968A (en) * | 2003-06-03 | 2006-07-05 | 尤纳克西斯巴尔策斯公司 | Optical substrate for enhanced detectability of fluorescence |
| CN1837890A (en) * | 2006-03-29 | 2006-09-27 | 哈尔滨工业大学 | Image recovery and pupil filtering type transverse super-resolution confocal microscopic imaging method and apparatus |
| CN101013136A (en) * | 2007-02-08 | 2007-08-08 | 北京工业大学 | Laser-induction fluorescence co-focusing scan device and method |
| US20090213376A1 (en) * | 2008-02-22 | 2009-08-27 | Olympus Corporation | Optical equipment |
| EP2312367A1 (en) * | 2009-10-16 | 2011-04-20 | Olympus Corporation | Laser scanning microscope |
| CN201917521U (en) * | 2010-12-17 | 2011-08-03 | 北京柏奥达思科技有限公司 | Confocal fluorescent detection device and immunity bedside detector adopting same |
-
2012
- 2012-03-09 CN CN201210061262.XA patent/CN102621117B/en not_active Expired - Fee Related
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0538551A2 (en) * | 1991-10-24 | 1993-04-28 | Toa Medical Electronics Co., Ltd. | Flow imaging cytometer |
| WO1994029695A1 (en) * | 1993-06-08 | 1994-12-22 | Gjelsnes Oddbjoern | Liquid flow cytometer |
| WO1999037999A1 (en) * | 1998-01-27 | 1999-07-29 | Wisconsin Alumni Research Foundation | Signal enhancement for fluorescence microscopy |
| JP2000227556A (en) * | 1999-02-05 | 2000-08-15 | Japan Science & Technology Corp | Microscope |
| CN1798968A (en) * | 2003-06-03 | 2006-07-05 | 尤纳克西斯巴尔策斯公司 | Optical substrate for enhanced detectability of fluorescence |
| JP2006011045A (en) * | 2004-06-25 | 2006-01-12 | Olympus Corp | Total reflection microscope |
| CN1605856A (en) * | 2004-11-18 | 2005-04-13 | 上海交通大学 | Laser fluorescence correlation spectrum unimolecular analyzer |
| CN1837890A (en) * | 2006-03-29 | 2006-09-27 | 哈尔滨工业大学 | Image recovery and pupil filtering type transverse super-resolution confocal microscopic imaging method and apparatus |
| CN101013136A (en) * | 2007-02-08 | 2007-08-08 | 北京工业大学 | Laser-induction fluorescence co-focusing scan device and method |
| US20090213376A1 (en) * | 2008-02-22 | 2009-08-27 | Olympus Corporation | Optical equipment |
| EP2312367A1 (en) * | 2009-10-16 | 2011-04-20 | Olympus Corporation | Laser scanning microscope |
| CN201917521U (en) * | 2010-12-17 | 2011-08-03 | 北京柏奥达思科技有限公司 | Confocal fluorescent detection device and immunity bedside detector adopting same |
Cited By (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102944540A (en) * | 2012-10-11 | 2013-02-27 | 中国科学院西安光学精密机械研究所 | Three-dimensional imaging system and method in deep scattering medium |
| CN103033920A (en) * | 2012-12-03 | 2013-04-10 | 苏州大猫单分子仪器研发有限公司 | Far infrared temperature jump microscope |
| CN103033920B (en) * | 2012-12-03 | 2015-04-08 | 苏州大猫单分子仪器研发有限公司 | Far infrared temperature jump microscope |
| CN104181089A (en) * | 2013-05-22 | 2014-12-03 | 中国石油化工股份有限公司 | Equipment for scanning facial porosity of rock and method thereof |
| CN104178420A (en) * | 2014-09-05 | 2014-12-03 | 中国科学技术大学 | Living cell culture environment control system for microscope |
| CN104178420B (en) * | 2014-09-05 | 2016-04-13 | 中国科学技术大学 | A kind of microscope viable cell culture environment Controlling System |
| CN104535547A (en) * | 2014-11-29 | 2015-04-22 | 北京工业大学 | Confocal optical path portable apparatus for observing micro-fluidic chip |
| CN104535547B (en) * | 2014-11-29 | 2017-04-05 | 北京工业大学 | A kind of common focused light passages portable unit of observation micro-fluidic chip |
| CN107683430B (en) * | 2015-06-02 | 2021-01-01 | 生命技术公司 | System and method for generating structured illumination images |
| CN107683430A (en) * | 2015-06-02 | 2018-02-09 | 生命技术公司 | System and method for generating structureization illumination image |
| CN107688014A (en) * | 2016-09-21 | 2018-02-13 | 北京大学 | A kind of cell imaging method |
| CN106568755A (en) * | 2016-11-06 | 2017-04-19 | 浙江大学 | Near infrared laser scanning confocal microscopic imaging system |
| CN111065950A (en) * | 2017-09-15 | 2020-04-24 | 敏捷焦点设计有限责任公司 | Dynamic focus and zoom system for wide-field, confocal and multiphoton microscopes |
| CN111065950B (en) * | 2017-09-15 | 2022-05-03 | 敏捷焦点设计有限责任公司 | Dynamic focus and zoom system for wide-field, confocal and multiphoton microscopes |
| CN107702661A (en) * | 2017-09-19 | 2018-02-16 | 福建师范大学 | A kind of white light self-interference Surface testing instrument |
| CN109355177A (en) * | 2018-11-29 | 2019-02-19 | 苏州度奕星科技有限公司 | A kind of living cells monitoring device |
| WO2021120509A1 (en) * | 2019-12-18 | 2021-06-24 | 江南大学 | Instrument and method for counting microbial cells |
| US11346763B2 (en) | 2019-12-18 | 2022-05-31 | Jiangnan University | Apparatus and method for microbial cell counting |
| CN111258044A (en) * | 2020-02-18 | 2020-06-09 | 清华大学 | Mirror scanning light field-based 4 pi-like microscopic imaging method |
| CN112268505A (en) * | 2020-10-22 | 2021-01-26 | 江苏集萃微纳自动化系统与装备技术研究所有限公司 | Microscopic operation system based on full-field optical coherence tomography technology |
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