CN1362048A - Visible light induced delayed lighting no-damage tumor imaging method and equipment - Google Patents
Visible light induced delayed lighting no-damage tumor imaging method and equipment Download PDFInfo
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Abstract
本发明提供一种可见光诱导的延迟发光无损伤肿瘤成像方法,包括下述步骤:卟啉类光敏试剂和延迟发光试剂活性血清白蛋白的预先导入及可见光直接照射生物体或组织,诱导生物体或组织产生较长时间的内源性延迟发光;光接收组件将此发光信号转换为电信号;电信号通过模数转换为数字信号;处理数字信号获得生物体或组织的结构图像;一种可见光诱导的延迟发光无损伤肿瘤成像装置,包括有可见光激发组件、光接收组件、模数转换器、计算机。本发明结合了延迟发光的背景干扰低及光学成像具有高分辨、无损伤的优点,且所使用的试剂对生物体无化学损伤,对肿瘤病变部位有选择性聚集的特点,用于生物医学中肿瘤病变的影像诊断得到较高分辨率和清晰的图像。
The present invention provides a visible light-induced delayed luminescence non-damaging tumor imaging method, comprising the following steps: pre-introducing porphyrin-based photosensitizers and delayed luminescence reagent active serum albumin and directly irradiating organisms or tissues with visible light to induce organisms or The tissue produces endogenous delayed luminescence for a long time; the light-receiving component converts the luminescence signal into an electrical signal; the electrical signal is converted into a digital signal through analog-to-digital conversion; the digital signal is processed to obtain a structural image of the organism or tissue; a visible light-induced The delayed luminescence non-invasive tumor imaging device includes a visible light excitation component, a light receiving component, an analog-to-digital converter, and a computer. The invention combines the advantages of low background interference of delayed luminescence and high resolution and no damage of optical imaging, and the reagents used have no chemical damage to organisms, and have the characteristics of selective aggregation of tumor lesion sites, and are used in biomedicine. Imaging diagnosis of tumor lesions obtains higher resolution and clearer images.
Description
(1) technical field
The present invention relates to a kind of medical imaging diagnosis technology of tumor, the method and the device thereof of damage tumor imaging carried out in particularly a kind of delay chemiluminescence that utilizes visible light-inducing.
(2) background technology
At present in the imaging technique that medical imaging diagnosis generally uses, X ray transmission imaging, x-ray tomography (CT), NMR (Nuclear Magnetic Resonance)-imaging etc. all have largely damage to tissue, and the cost of x-ray tomography instrument, nuclear magnetic resonance imaging instrument is relatively more expensive; B-mode ultrasonography does not have damage substantially to human body, and cost is also lower, but because the principle of B-mode ultrasonography is organizational interface's catoptric imaging, so can't realize chromatography, and resolution lower (to in-vivo tissue simple in structure, best resolution is several millimeters), contrast is also relatively poor.Optical image technology (comprises time discrimination optics imaging, the domain optical imaging, optical coherence tomography etc.) have the human zero damage, characteristics such as resolution height, but because these methods all are to utilize external light source, and biological tissue has intensive scattering and absorption to external source light, so present technical merit can only reach the shallow-layer biological tissue imaging of the degree of depth for the millimeter magnitude, the difficult accurate measurement that realizes deep tissues, simultaneously, these optical imaging methods all utilize optical properties of tissue to realize, its image contrast derives from optical properties of tissue difference only singlely, thereby image contrast is not high.
(3) summary of the invention
The object of the present invention is to provide a kind of delayed lighting no-damage tumor imaging method of visible light-inducing.Mainly be to use two kinds of reagent harmless to organism, in conjunction with short time irradiation to the harmless visible light of organism, the endogenous of inducing organism or tissue to produce the long period postpones chemiluminescence, and utilizes this endogenous delayed lighting no-damage ground that accurately image is realized at organism neoplastic lesion position.
Another object of the present invention is to provide a kind of delayed lighting no-damage tumor imaging device of realizing the visible light-inducing of said method.
Purpose of the present invention is achieved through the following technical solutions, and the delayed lighting no-damage tumor imaging method of this visible light-inducing comprises the steps:
(1) will be to organism or the no chemical damage of tissue, and the neoplastic lesion position is had the specific porphyrin class of metabolism photosensitive reagents 1～100 μ M be injected into organism or tissue;
After (2) 10～24 hours, will have active serum albumin 1～100 μ M and be injected into organism or tissue as delayed luminescence reagent;
(3) be 0.05～2.00W/cm with power density
2Be the even irradiated biological body of excited by visible light light source of 420～700nm or organize 10～60s that the endogenous of inducing organism or tissue to produce the long period postpones chemiluminescence with wave-length coverage;
(4) utilizing optical fiber receive module to receive this endogenous from organism or tissue postpones chemiluminescence signal and is converted into the signal of telecommunication;
(5) be digital signal with the signal of telecommunication by analog digital conversion;
(6) processing digital signal and carry out image reconstruction obtains the structural images of organism or tissue.
The wavelength of described excitation source can be selected to be in the absorbing wavelength scope of photosensitizer, and to organism or organize 500～700nm of the better penetrance of tool; Excitation light power density can be used the amount 0.1～0.5W/cm far below biological tissue's damage threshold
2
The endogenous of inducing organism or tissue to produce the long period at even irradiated biological body or tissue with excitation source postpones to close excitation source after the chemiluminescence, and darkening 1～10s can avoid the interference from the fluorescence and the phosphorescence of body from biological tissue like this; Because of delayed luminescence decay fast in time, shorten the darkening time, can improve the delayed luminescence signal intensity.
Described excitation source can be laser uniform irradiation biological sample to be measured after expanding bundle of single wavelength, also can be with the mercury lamp with broad bands of a spectrum or xenon lamp as excitation source uniform irradiation biological sample to be measured, be the damage of the ultraviolet band that prevents light source, can select for use suitable optical filter to filter ultraviolet light in the exciting light biological tissue.
When described excitation source is the high voltage mercury lamp of 50W, chooses the long pass filter of 420nm and 495nm and obtain the required wave band that excites, vertical evenly irradiation sample 10～60s can obtain the delayed luminescence image of organism tumor locus clearly; Signal to noise ratio can reach 1.42.
The delayed lighting no-damage tumor imaging device of visible light-inducing of the present invention comprises excited by visible light assembly, optical fiber receive module, analog-digital converter, computer, light excitation component and computer are electrically connected, and optical fiber receive module, analog-digital converter and computer are electrically connected successively.
Described excited by visible light assembly can adopt the laser of single wavelength, also can be with the normal excitation source that uses, for example xenon lamp or mercury lamp in the existing fluorescence imaging.
Described optical fiber receive module is the device that is used to survey light, and is comparatively faint comparatively speaking because of delayed luminescence, can adopt the single photon imaging system, for example CCD or the enhancement mode CCD system that takes pictures.
Described analog-digital converter can adopt general analog-digital converter, or has other devices of analog-digital conversion function.The CCD controller that for example image series capture card of National Instruments company production, or PrincetonInstruments company is produced.
General image treatment software is housed in the described computer, for example the WINVIEW software of the LabVIEW software of Nationa1 Instruments company exploitation or the exploitation of Princeton Instruments company is used for organism or organizes delayed luminescence treatment of picture and reconstruction.
The present invention compared with prior art has following advantage and effect:
(1) delayed lighting no-damage tumor imaging method of this visible light-inducing combines that radiation-induced endogenous of short time of visible light postpones chemiluminescence and optical imagery has high-resolution, undamaged advantage, can not produce radiation injury to organism, thus compare with traditional medical imaging diagnosis method have not damaged, advantage that resolution is high.
(2) delayed lighting no-damage tumor imaging method of this visible light-inducing can be penetrated biological tissue after a period of time at exciting light irradiation, close exciting light and darkening certain hour, and then irradiated biological tissue carried out the delayed luminescence imaging, so this formation method has thoroughly avoided coming from the interference of biological tissue from the fluorescence and the phosphorescence of body, with traditional fluorescence imaging method, for example the photodynamics fluorescence diagnosis is compared and is greatly reduced ambient interferences, and contrast also is greatly increased.
(3) two kinds of reagent of the delayed lighting no-damage tumor imaging method of this visible light-inducing use, porphyrin class photosensitive reagents and serum albumin, can not produce various chemical damages, so compare advantage with not damaged living imaging with the various fluorescence contrast compositions and methods of traditional use to organism.
(4) the delayed lighting no-damage tumor imaging method operating process of this visible light-inducing is simple, realizes easily; And the apparatus structure of realizing this method is simple, and the production assembling is more or less freely, easy for operation, and the single unit system cost is relatively low.
(4) description of drawings
Fig. 1 is the structural representation of the delayed lighting no-damage tumor imaging device of visible light-inducing of the present invention;
Fig. 2 contains in-vitro simulated solution in a cuvette writes down 30s behind excited by visible light delayed luminescence image;
Fig. 3 is the outline drawing of a right shoulder tumor bearing nude mice under faint light is shone;
Fig. 4 be the tumor bearing nude mice of Fig. 3 by excited by visible light after the delayed luminescence image of record 1min.
(5) specific embodiment
Below in conjunction with accompanying drawing the present invention is done further concrete description, but embodiments of the present invention are not limited thereto.
Embodiment 1
Fig. 1 shows the structure of the delayed lighting no-damage tumor imaging device of visible light-inducing of the present invention, as seen from Figure 1, this device includes light excitation component 1, optical fiber receive module 2, analog-digital converter 3, computer 4, and wherein light excitation component 1 is made of excitation source 1-1, focusing lens 1-2 and filter set 1-3; Optical fiber receive module 2 is connected and composed by take pictures camera lens 2-1 and the detector 2-2 that collect light; Light excitation component 1 is electrically connected with computer 4, and optical fiber receive module 2, analog-digital converter 3 are electrically connected successively with computer 4; 5 is three-dimensional adjustable specimen holder, places organism to be measured or tissue on the frame, and 6 is the darkroom.Select for use each member to connect to form this device, wherein, the common 50W high voltage mercury lamp that excitation source 1-1 selects for use BIO-RAD company to make, focusing lens 1-2 is the pancratic lens group that mercury lamp carries, GG 420 and GG 495 long pass filters that optical filter 1-3 selects for use COHERENT company to make; The camera lens 2-1 that takes pictures selects the big digital aperture camera lens (50mm, f/1.2 type) of the Nikon company manufacturing that is applicable to that the low light level is taken for use; The ICCD-576-S/1 enhancement mode CCD that detector 2-2 selects for use U.S. Princeton Instruments company to make; The ST-130 type controller that analog-digital converter 3 selects for use U.S. Princeton Instruments company to make; Computer 4 is selected the Pentium III type microcomputer of Intel Company for use.
Said apparatus is applied to the imaging of in-vitro simulated solution: in the quartz colorimetric utensil of a 10mm * 10mm * 42mm, add the neutral normal saline of 1mL, wherein contain the protoporphyrin IX of 15 μ M and the active bovine serum albumin of 20 μ M, after being mixed, solution is positioned on the specimen holder 5 in the darkroom 6, make liquid level be positioned at the take pictures focal plane place of camera lens 2-1 of light harvesting, close darkroom 6 then fully, open be in the light shutter and the on and off switch of excitation source 1-1 by the control software of computer 4, make exciting light pass through focusing lens 1-2 vertical with optical filter 1-3 and equably irradiation contain a quartz window face of sample cuvette, by the shutter that is in the light of closing excitation source behind the computer 4 control exposure time 20s, under the excitation of light, the endogenous that this in-vitro simulated solution can produce about 5min postpones chemiluminescence, for avoiding possible fluorescence and phosphorescence to disturb (the triplet excited state life-span that generally all has the long period because of photosensitizer, so may produce the phosphorescence of longer life), close being in the light soon behind the door of excitation source, allow testing sample darkening 2s in darkroom 6, open the take pictures shutter of camera lens 2-1 of light harvesting by computer 4 control then, by the delayed luminescence signal of detector 2-2 opening entry 30s from sample, and luminous signal is converted into the signal of telecommunication, 3 pairs of signals of telecommunication of analog-digital converter carry out analog digital conversion be behind the digital signal with its input computer 4, in computer 4, utilize the WINVIEW software of Princeton Instruments company exploitation to carry out image reconstruction and date processing and can obtain as shown in Figure 2 the delayed luminescence image of in-vitro simulated solution under the excited by visible light effect.As seen from Figure 2, the inventive method can be to in-vitro simulated biological tissue's solution imaging, and clear picture, signal to noise ratio are 2.76.
Embodiment 2
Device described in the embodiment 1 is applied to imaging to biological living: get 4 monthly ages experiments with one of nude mice, in the right shoulder subcutaneous vaccination of nude mice lung adenocarcinoma cell, two week the back inoculation positions grow the solid tumor of the about 1cm size of diameter.Give the dosage intravenous injection Porphyrin-Based Sensitizer chlorin e4 of this nude mice the previous day of experiment by 1～10mg/Kg body weight, the active serum albumin of subcutaneous injection 20～200mg/Kg body weight after 24 hours, behind the 60min nude mice is anaesthetized, put on the specimen holder 5 in darkroom 6, at first under shining, faint light writes down outline drawing as shown in Figure 3, orient the orientation of Mus body and tumor, close darkroom 6 then fully, open be in the light shutter and the power supply of excitation source 1-1 by computer 4, make exciting light can be vertically irradiation nude mice surface equably, by computer 4 control exposure times 10～60s, close the shutter that is in the light of excitation source 1-1 then, under the excitation of light; the endogenous that the tumor locus of pathological changes produces about 5min postpones chemiluminescence; disturb (the triplet excited state life-span that generally all has the long period because of photosensitizer; so may produce the phosphorescence of longer life) for avoiding possible fluorescence and phosphorescence; close being in the light soon behind the door of excitation source 1-1; allow nude mice to be measured darkening 1～10s in darkroom 6 in; open the take pictures shutter of camera lens 2-1 of light harvesting by computer 4 controls then; postpone chemiluminescence signal by detector 2-2 opening entry 1min from the endogenous of nude mice; and luminous signal is converted into the signal of telecommunication; it is behind the digital signal it to be imported computer 4 that 3 pairs of signals of telecommunication of analog-digital converter carry out analog digital conversion, the WINVIEW software that utilizes Princeton Instruments company to develop in computer 4 carries out image reconstruction and date processing can obtain as shown in Figure 4 delayed luminescence image.As seen from Figure 4, the inventive method can be to the neoplastic lesion position imaging of biological living, and image is comparatively clear, and signal to noise ratio is 1.42.
Claims (9)
1, a kind of delayed lighting no-damage tumor imaging method of visible light-inducing is characterized in that comprising the steps:
(1) will be to organism or the no chemical damage of tissue, and the neoplastic lesion position is had the specific porphyrin class of metabolism photosensitive reagents 1～100 μ M be injected into organism or tissue;
After (2) 10～24 hours, 1～100 μ M is injected into organism or tissue as delayed luminescence reagent with the active serum albumin;
(3) be 0.05～2.00W/cm with power density
2Be the even irradiated biological body of excited by visible light light source of 420～700nm or organize 10～60s that the endogenous of inducing organism or tissue to produce the long period postpones chemiluminescence with wave-length coverage;
(4) utilizing optical fiber receive module to receive this endogenous from organism or tissue postpones chemiluminescence signal and is converted into the signal of telecommunication;
(5) be digital signal with the signal of telecommunication by analog digital conversion;
(6) processing digital signal and carry out image reconstruction obtains the structural images of organism or tissue.
2, the delayed lighting no-damage tumor imaging method of visible light-inducing according to claim 1 is characterized in that: described tumor locus is had concentration range 30～50 μ M and the delayed luminescence reagent that the specific Porphyrin-Based Sensitizer of metabolism is clinical permission is that 30～50 μ M have active serum albumin.
3, the delayed lighting no-damage tumor imaging method of visible light-inducing according to claim 1, it is characterized in that: described excitation source can be laser uniform irradiation biological sample to be measured after expanding bundle of single wavelength, also can be with the mercury lamp with broad bands of a spectrum or xenon lamp as excitation source uniform irradiation biological sample to be measured.
4, the delayed lighting no-damage tumor imaging method of visible light-inducing according to claim 3, it is characterized in that: when excitation source is the high voltage mercury lamp of 50W, can choose the long pass filter of 420～500nm and obtain the required wave band that excites, vertical evenly irradiation sample 10～60s.
5, the delayed lighting no-damage tumor imaging method of visible light-inducing according to claim 1, it is characterized in that: the wavelength of described excitation source is to be in the absorption region of photosensitizer, and to organism or organize 500～700nm of the better penetrance of tool.
6, the delayed lighting no-damage tumor imaging method of visible light-inducing according to claim 1 is characterized in that: the excitation source power density can be used the amount 0.1～0.5W/cm far below biological tissue's damage threshold
2
7, the delayed lighting no-damage tumor imaging method of visible light-inducing according to claim 1, after it is characterized in that even irradiated biological body of described excitation source or tissue, close excitation source, darkening 1～10s, the delayed luminescence of the organism of opening entry visible light-inducing or tissue then.
8, a kind of delayed lighting no-damage tumor imaging device of visible light-inducing, it is characterized in that comprising excited by visible light assembly, optical fiber receive module, analog-digital converter, computer, light excitation component and computer are electrically connected, and optical fiber receive module, analog-digital converter and computer are electrically connected successively.
9, the delayed lighting no-damage tumor imaging device of visible light-inducing according to claim 8 is characterized in that: described optical fiber receive module adopts the single photon imaging system.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1778344B (en) * | 2004-11-26 | 2010-05-12 | 天津天士力制药股份有限公司 | drugs for coronary heart disease |
| CN1778345B (en) * | 2004-11-26 | 2010-05-12 | 天津天士力制药股份有限公司 | Chinese Medicine Dropping Pills for Treating Coronary Heart Disease |
| CN109142305A (en) * | 2015-02-16 | 2019-01-04 | 北京大学 | Living animal two-photon excitation delay detection fluorescence imaging analysis method and apparatus |
| CN110108677A (en) * | 2019-05-07 | 2019-08-09 | 新绎健康科技有限公司 | Biological delayed luminescence detection system |
| CN111000531A (en) * | 2019-12-26 | 2020-04-14 | 新绎健康科技有限公司 | Method and device for evaluating body changes before and after exercise |
| CN115607110A (en) * | 2022-10-18 | 2023-01-17 | 广州大学 | A breast tumor detection system based on autofluorescence |
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2002
- 2002-01-24 CN CN 02114781 patent/CN1362048A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1778344B (en) * | 2004-11-26 | 2010-05-12 | 天津天士力制药股份有限公司 | drugs for coronary heart disease |
| CN1778345B (en) * | 2004-11-26 | 2010-05-12 | 天津天士力制药股份有限公司 | Chinese Medicine Dropping Pills for Treating Coronary Heart Disease |
| CN109142305A (en) * | 2015-02-16 | 2019-01-04 | 北京大学 | Living animal two-photon excitation delay detection fluorescence imaging analysis method and apparatus |
| CN109142305B (en) * | 2015-02-16 | 2021-12-14 | 北京大学 | Two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment for living animals |
| CN110108677A (en) * | 2019-05-07 | 2019-08-09 | 新绎健康科技有限公司 | Biological delayed luminescence detection system |
| CN111000531A (en) * | 2019-12-26 | 2020-04-14 | 新绎健康科技有限公司 | Method and device for evaluating body changes before and after exercise |
| CN115607110A (en) * | 2022-10-18 | 2023-01-17 | 广州大学 | A breast tumor detection system based on autofluorescence |
| CN115607110B (en) * | 2022-10-18 | 2023-11-21 | 广州大学 | Mammary gland tumor detection system based on autofluorescence |
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