CN105548102A - Deep calcium ion concentration monitoring method using photoacoustic-fluorescent complementary principle and device using the same - Google Patents
Deep calcium ion concentration monitoring method using photoacoustic-fluorescent complementary principle and device using the same Download PDFInfo
- Publication number
- CN105548102A CN105548102A CN201510896712.0A CN201510896712A CN105548102A CN 105548102 A CN105548102 A CN 105548102A CN 201510896712 A CN201510896712 A CN 201510896712A CN 105548102 A CN105548102 A CN 105548102A
- Authority
- CN
- China
- Prior art keywords
- calcium ion
- ion concentration
- optoacoustic
- signal
- fluorescence
- 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.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/1702—Systems in which incident light is modified in accordance with the properties of the material investigated with opto-acoustic detection, e.g. for gases or analysing solids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/1702—Systems in which incident light is modified in accordance with the properties of the material investigated with opto-acoustic detection, e.g. for gases or analysing solids
- G01N2021/1706—Systems in which incident light is modified in accordance with the properties of the material investigated with opto-acoustic detection, e.g. for gases or analysing solids in solids
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a deep calcium ion concentration monitoring method using a photoacoustic-fluorescent complementary principle and a device using the same. The method can detect calcium ion concentration change through detection of change of intensity of a photoacoustic signal of a calcium ion fluorescent probe. The device comprises a photoacoustic-fluorescent signal excitation source, a bowl-shaped hollow focusing ultrasonic detector, a photomultiplier, a reflector, a light splitter, an expanded beam lens group, a microobjective, an ultrasonic coupling beaker, a sample bench, a 3D scanning platform, an optical filter, a signal amplifier, a two-channel parallel acquisition card and a computer with image reconstruction software, acquisition control software and motor control software. The method and device realize deep tissue calcium ion concentration monitoring, are convenient for operation, has calcium ion fluorescent probe sensitivity of 0.15 micromoles per liter and realizes monitoring of change of a deep tissue calcium ion concentration. The method and device can realize detection of change of a deep tissue calcium ion concentration in real time and through the rate of the change, a pathological change of tissues can be deduced.
Description
Technical field
The invention belongs to non-destructive testing field of measuring technique, particularly a kind of deep layer calcium ion concentration monitoring method and device utilizing optoacoustic-fluorescence complementary principle.
Background technology
Fluorescence signal at present mainly by observing calcium ion fluorescent monitors the change of calcium ion concentration.But because biological tissue is on the impact of fluorescent scattering, fluorescence imaging method can not monitor the change of deep layer calcium ion concentration.And photoacoustic signal (photic ultrasonic) by the impact of tissue scatter much smaller than the impact suffered by photon, therefore deep penetration characteristic in harmless photoacoustic imaging technology and pure optical imagery high sensitivity characteristic and pure ultrasonic imaging combines by we, overcome light scattering restriction, achieve the high-resolution to live body deep tissues, high-contrast image.Calcium ion is the indispensable ion of the every physiological activity of body.It, for the biopotential maintaining cell membrane both sides, maintains normal Nerve conduction, maintains normal muscle and stretches and diastolic function and nerve, muscle conduction function, play very important effect.Calcium ion, as the first messenger in signal transduction pathway, the synaptic neural for whole central nervous system is first and neuroglia is most important.
Summary of the invention
In order to overcome the shortcoming of prior art with not enough, primary and foremost purpose of the present invention is to provide a kind of deep layer calcium ion concentration monitoring method utilizing optoacoustic-fluorescence complementary principle.
Another object of the present invention is to provide the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle.
Object of the present invention is achieved through the following technical solutions:
Utilize a deep layer calcium ion concentration monitoring method for optoacoustic-fluorescence complementary principle, comprise the steps:
(1) utilize calcium ion fluorescent as optoacoustic calcium ion probe;
Because the fluorescence intensity of calcium ion fluorescent can strengthen along with the increase of the concentration of calcium ion or weaken.The optoacoustic produced in same excitation process, fluorescence signal are shifting, there is complementary relationship.Therefore, the photoacoustic signal of calcium ion fluorescent can increase along with the concentration of calcium ion and reduce or strengthen.Therefore, by the change of the photoacoustic signal power of detection calcium ion fluorescent, the change of calcium ion concentration size can be detected.Again because photoacoustic waves (photoacoustic signal) is little more than fluorescence by the impact of tissue scatter, therefore utilize optoacoustic-fluorescence complementary principle, the change of deep tissues calcium ion can be monitored by the photoacoustic signal detecting calcium ion fluorescent.
(2) calcium ion fluorescent is expelled to the region intending detecting;
(3) pulse laser that sends of optoacoustic fluorescence signal excitaton source is after microcobjective, focuses on surveyed area, makes calcium ion fluorescent produce photoacoustic signal and fluorescence signal;
(4) photoacoustic signal is received by bowl-shape hollow focus supersonic detector through after ultrasonic coupling liquid, then amplifies through signal amplifier; Fluorescence signal is received by photomultiplier after spectroscope and optical filter filter; Two paths of signals is gathered by binary channels parallel acquisition card simultaneously, then data is transmitted and be stored into gathering in the computing machine of control software design and image reconstruction software;
(5) detect sample is positioned on three dimensional scanning platform together with the ultrasonic coupling cup holding ultrasonic coupling liquid, is moved by the three-dimensional of the computing machine Quality control that motor control software is housed; Often gathered the photoacoustic signal of a point, three dimensional scanning platform horizontal direction is moved in order and is moved a step; After having scanned a plane, platform has vertically moved a step again; To carry out point by point scanning, for reconstruction of three-dimensional images to sample; Final acquisition three-dimensional light acoustic signal intensity distribution, to obtain calcium ion concentration distribution;
(6) photoacoustic signal of calcium ion fluorescent collected after, reconstruct calcium ion probe light acoustic signal intensity distributed image by maximum value projection algorithm, judge the concentration change of calcium ion according to photoacoustce signal intensity;
(7) by the photoacoustce signal intensity of the calcium ion fluorescent of detection single-point, the change of this calcium ion concentration of Real-Time Monitoring.
Calcium ion fluorescent described in step (1) is preferably Rhod-2, but does not limit above-mentioned probe.
Calcium ion fluorescent selected in described step (2) with calcium binding before and after absorption spectrum without significant change.
Optoacoustic fluorescence signal excitaton source described in step (3) is pulsed laser, and Output of laser wavelength is selected according to the excitation wavelength of calcium ion fluorescent.
The laser instrument of to be output laser pulse width be nanosecond or the psec of the pulsed laser described in step (3), output wavelength is 532nm, 675 ~ 1000nm, repetition frequency 20Hz ~ 10KHz.
Motor control software described in step (4) is Labview software, and program is write voluntarily.
Ultrasonic coupling liquid described in step (5) is water;
The dominant frequency of the bowl-shape hollow focus supersonic detector described in step (5) is 10MHz, hollow focusing probe.
Collection control software design described in step (5) and image reconstruction software are Labview software, and program is write voluntarily;
Maximum value projection algorithm described in step (6) is write voluntarily by Matlab software.
Detection single-point described in step (7), utilizes the pulsed laser of high repetition frequency (10KHz) and the capture card 200M/s of high sampling rate.
Utilize the deep layer calcium ion concentration monitoring device of optoacoustic-fluorescence complementary principle, build according to said method.Described pick-up unit comprise optoacoustic fluorescence signal excitaton source, bowl-shape hollow focus supersonic detector, photomultiplier, catoptron, spectroscope, extender lens group, microcobjective, ultrasonic coupling cup, sample stage, three dimensional scanning platform, optical filter, signal amplifier, binary channels parallel acquisition card, with the computing machine gathering control software design and image reconstruction software and motor control software.
The pulsed laser of described optoacoustic fluorescence signal excitaton source to be output laser pulse width be nanosecond or psec, transmit light on microcobjective through catoptron after Laser output, microcobjective focuses light on sample.
The bowl-shape hollow ultrasonic probe of described bowl-shape hollow focus supersonic detector is fixed on below microcobjective, makes microcobjective coaxial with ultrasonic probe, and luminous energy focuses on the focus of ultrasonic probe through probe from the aperture in the middle of probe, realize light, sound is confocal.The photoacoustic signal of generation is excited to be received by ultrasonic probe.
It is inner that described microcobjective is fixed on bowl-shape hollow focus supersonic detector, and ensure microcobjective and the confocal point of bowl-shape hollow focus supersonic detector; The described ultrasonic coupling cup holding ultrasonic coupling liquid is positioned at directly over sample, and bowl-shape hollow focus supersonic detector and photomultiplier are placed in directly over ultrasonic coupling cup, and wherein photomultiplier is above bowl-shape hollow focus supersonic detector;
Described optoacoustic fluorescence excitation light source, signal amplifier, photomultiplier, three dimensional scanning platform, binary channels parallel acquisition card be electrically connected successively with the computing machine gathering control software design and image reconstruction software and motor control software;
The enlargement factor of described signal amplifier is 70dB, and bandwidth is 50KHz ~ 500MHz;
Described signal amplifier is preferably multi-stage cascade amplifier;
Described microcobjective is flat-field objective, and enlargement factor is 4 times, 10 times, 20 times, 40 times, 100 times, and image quality is only relevant with enlargement factor, and the higher then quality of multiple is better.
The model of described binary channels parallel acquisition card can be PCI2400 (production of NI company), but other models, such as grind magnificent capture card and also can use.
Described three dimensional scanning platform is made up of two-dimentional electric platforms and lifting table; Described two-dimentional electric platforms is made up of stepper motor and mobile platform, and rotarily drive mobile platform by motor and move forward and backward, two-dimentional electric platforms is made up of two this electric platforms, can realize four direction all around and move; Control with the effect reaching Different Plane primarily of lifting table at vertical direction.
Described collection control software design and image reconstruction software are Labview software, and program is write voluntarily;
Described motor control software is Labview software, and program is write voluntarily; Imaging software can be compiled with Matlab software and write voluntarily.
Principle of the present invention is: the process based on optoacoustic-fluorescence complementary principle and calcium ion fluorescent stimulated radiation only has two approach: radiation transistion and nonradiative transition.Fluorescence imaging and photoacoustic imaging realize the detection of the Conversion of Energy form (fluorescence and optoacoustic) produced in these two approach just.Fluorescence imaging detection be the energy (fluorescence) of radiation transistion, and photoacoustic imaging detection is the energy (photoacoustic signal) of nonradiative transition.For general calcium ion fluorescent, the radiation transistion after being excited and nonradiative transition process are all occur simultaneously and there is the mechanism of vying each other, and both actual occurrence probability are then determined by the concentration of calcium ion.Usual calcium ion concentration has fluorescence signal intensity to characterize.Because optoacoustic, fluorescence exist shifting complementary relationship, photoacoustic signal equally also can reflect the concentration change of calcium ion.And the photoacoustic signal transmission degree of depth is in the tissue firmly got nearly to 1cm than fluorescence.Therefore, the monitoring that optoacoustic-fluorescence complementary principle can realize deep layer calcium ion concentration is utilized.This method will be used for the research of Neuscience.
The present invention, relative to prior art, has following advantage and effect:
Method and apparatus of the present invention can realize the calcium ion concentration monitoring of deep tissues; Simple to operate, the sensitivity of monitoring calcium ion fluorescent can reach 0.15 μM, can monitor the change of deep tissues calcium ion concentration.The present invention can detect the situation of change of deep tissues calcium ion concentration in real time, can infer tissue pathologic change degree thus by the speed of this change.
Accompanying drawing explanation
Fig. 1 is the structural representation of the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle; Wherein, 1 is the computing machine with gathering control software design and image reconstruction software and motor control software; 2 is three dimensional scanning platform; 3 is optoacoustic fluorescence signal excitaton source; 4 is catoptron; 5 for placing the sample stage of sample; 6 is ultrasonic coupling cup; 7 is bowl-shape hollow focus supersonic detector; 8 is microcobjective; 9 is spectroscope; 10 is optical filter; 11 is photomultiplier; 12 is signal amplifier; 13 is extender lens group.
Fig. 2 is the optoacoustic of calcium ion concentration and calcium ion probe, the graph of a relation of fluorescence signal intensity.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
Utilize the structural representation of the deep layer calcium ion concentration monitoring device of optoacoustic-fluorescence complementary principle as shown in Figure 1.
This monitoring device comprises: 1 is the computing machine with gathering control software design and image reconstruction software and motor control software; 2 is three dimensional scanning platform; 3 is optoacoustic fluorescence signal excitaton source; 4 is catoptron; 5 for placing the sample stage of sample; 6 is ultrasonic coupling cup; 7 is bowl-shape hollow focus supersonic detector; 8 is microcobjective; 9 is spectroscope; 10 is optical filter; 11 is photomultiplier; 12 is signal amplifier; 13 is extender lens group.
Wherein, optoacoustic fluorescence excitation light source 3, signal amplifier 12, photomultiplier 11, three dimensional scanning platform 2 and be electrically connected successively with the computing machine 1 gathering control software design and image reconstruction software and motor control software;
It is inner that described microcobjective 8 is fixed on bowl-shape hollow focus supersonic detector 7, and ensure microcobjective 8 and the confocal point of bowl-shape hollow focus supersonic detector 7; The described ultrasonic coupling cup 6 holding ultrasonic coupling liquid is positioned at directly over the sample stage 5 of placement sample, both are as a whole, be fixed on three dimensional scanning platform 2, three dimensional scanning platform 2 drives the sample stage 5 placing sample move together with ultrasonic coupling cup 6 and scan; Load ultrasonic coupling liquid water in ultrasonic coupling cup 6 to propagate for photoacoustic signal; Bowl-shape hollow focus supersonic detector 7 and photomultiplier 11 are placed in directly over ultrasonic coupling cup 6, and wherein photomultiplier 11 is above bowl-shape hollow focus supersonic detector 7;
The pulse laser that described optoacoustic fluorescent exciting source generator 3 sends is reflected by catoptron 4, then be radiated on sample after microcobjective 8 focuses on, namely the some hot spot that pulse laser is formed after focusing on drops on sample, the photoacoustic signal produced first is received by bowl-shape hollow focus supersonic detector 7 after ultrasonic coupling liquid, then amplifies through signal amplifier 12; The fluorescence signal produced is received by photomultiplier 11 after spectroscope 9 and optical filter 10, and fluorescence signal and photoacoustic signal are gathered by with the computing machine 1 gathering control software design and image reconstruction software and motor control software (Labview) simultaneously.
Three dimensional scanning platform 2 is moved by the three-dimensional of the computing machine Quality control with motor control software; Often gathered the photoacoustic signal of a point, three dimensional scanning platform 2 horizontal direction is moved in order and is moved a step; After having scanned a plane, platform has vertically moved a step again; To carry out point by point scanning, for reconstruction of three-dimensional images to sample; Final acquisition three-dimensional light acoustic signal intensity distribution, to obtain calcium ion concentration distribution; After photoacoustic signal is collected, is reconstructed the photoacoustce signal intensity distributed image of sample by maximum value projection algorithm (for being write voluntarily by Matlab software), judging the concentration change of calcium ion according to photoacoustce signal intensity;
Calcium ion fluorescent (the present invention is for Rhod-2) is expelled to the plan surveyed area on the sample stage 5 placing sample.Optoacoustic fluorescence signal excitaton source 3 is exported pulse laser and is reflected by catoptron 4, impinges perpendicularly in extender lens group 13.Light beam is after expanding, and the laser scattered is detected by photomultiplier 9 with the shake of calibration laser energy.Laser beam focuses on monitored area by microcobjective 5.Calcium ion fluorescent is produced photoacoustic signal and fluorescence signal after laser excitation, photoacoustic signal, by the super coupling liquid water coincidence inside ultrasonic coupling cup 6, is then received by the bowl-shape hollow focus supersonic detector 7 below microcobjective 5.Detecting sample is placed on three dimensional scanning platform 2, is moved by the three-dimensional of the computing machine Quality control that motor control software is housed.Often gathered the photoacoustic signal of a point, three dimensional scanning platform 2 horizontal direction is moved in order and is moved a step.After having scanned a plane, platform has vertically moved a step again.Final acquisition three-dimensional light acoustic signal intensity distribution, to reflect that calcium ion concentration distributes.Photoacoustic signal is through signal amplifier 8, and the collected card of data gathers, and is transferred to the computing machine 1 with gathering control software design and image reconstruction software and motor control software, processes, demonstrate the figure or image that can reflect calcium ion concentration to data.
Embodiment 2
Pick-up unit described in Application Example 1 realizes utilizing the deep layer calcium ion concentration of optoacoustic-fluorescence complementary principle to monitor.
(1) be placed in the ware that different calcium ion concentration solution is housed by the calcium ion fluorescent (for Rhod-2) of same concentrations, the fluorescence signal now detected strengthens along with the increase of calcium ion concentration.When after the thick chicken-breasted of ware upper cover last layer 1mm (imitated biological tissue), fluoroscopic imaging systems can't detect fluorescence signal.
(2) the optoacoustic fluorescence signal excitaton source of Rhod-2 adopts semiconductor pumped laser instrument, and output wavelength is 532nm, and pulsewidth is 10ns, and repetition frequency is 100Hz ~ 10KHz.The pulse laser that above-mentioned laser instrument produces, focuses on Rhod-2 through microcobjective, produces photoacoustic signal and fluorescence signal.Photoacoustic signal is received by bowl-shape hollow focus supersonic detector (dominant frequency is 10MHz).Photoacoustic signal transmits and is stored in computing machine after signal amplifier amplifies.The change of calcium ion concentration is gone out by software analysis.
(3) after the data gathered being processed, optoacoustic and the fluoroscopic image of sample is reconstructed by the algorithm of maximum value projection, then the photoacoustce signal intensity value relevant with calcium ion concentration can be obtained by the peak-to-peak value of the number of winning the confidence, as Fig. 2 photoacoustic signal and calcium ion concentration graph of a relation, and then can reflect the concentration change of calcium ion.As can be seen from Figure 2, optoacoustic, fluorescence signal are shifting, there is complementary relationship.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from Spirit Essence of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. utilize a deep layer calcium ion concentration monitoring device for optoacoustic-fluorescence complementary principle, it is characterized in that: this device comprise optoacoustic fluorescence signal excitaton source, bowl-shape hollow focus supersonic detector, photomultiplier, catoptron, spectroscope, extender lens group, microcobjective, ultrasonic coupling cup, sample stage, three dimensional scanning platform, optical filter, signal amplifier, binary channels parallel acquisition card, with the computing machine gathering control software design and image reconstruction software and motor control software;
It is inner that described microcobjective is fixed on bowl-shape hollow focus supersonic detector, and ensure microcobjective and the confocal point of bowl-shape hollow focus supersonic detector; The described ultrasonic coupling cup holding ultrasonic coupling liquid is positioned at directly over sample, and bowl-shape hollow focus supersonic detector and photomultiplier are placed in directly over ultrasonic coupling cup, and wherein photomultiplier is above bowl-shape hollow focus supersonic detector;
Described optoacoustic fluorescence excitation light source, signal amplifier, photomultiplier, three dimensional scanning platform, binary channels parallel acquisition card be electrically connected successively with the computing machine gathering control software design and image reconstruction software and motor control software.
2. ask the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle described in 1 according to right, it is characterized in that: described optoacoustic fluorescence signal excitaton source is pulsed laser, for the laser instrument that output laser pulse width is nanosecond or psec, output wavelength is 532nm, 675 ~ 1000nm, repetition frequency 20Hz ~ 10KHz.
3. the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle according to claim 1, is characterized in that: the enlargement factor of described signal amplifier is 70dB, and bandwidth is 50KHz ~ 500MHz.
4. the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle according to claim 1, is characterized in that: described signal amplifier is multi-stage cascade amplifier.
5. the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle according to claim 1, is characterized in that: described three dimensional scanning platform is made up of two-dimentional electric platforms and lifting table; Described two-dimentional electric platforms is made up of stepper motor and mobile platform, and rotarily drive mobile platform by motor and move forward and backward, two-dimentional electric platforms is made up of two this electric platforms, realizes four direction all around and moves; Control by lifting table the effect reaching Different Plane at vertical direction.
6. the deep layer calcium ion concentration monitoring device utilizing optoacoustic-fluorescence complementary principle according to claim 1, is characterized in that: described collection control software design and image reconstruction software are Labview software;
Described motor control software is Labview software.
7. use the monitoring method utilizing the deep layer calcium ion concentration monitoring device of optoacoustic-fluorescence complementary principle described in any one of claim 1 ~ 6, it is characterized in that comprising the steps:
(1) utilize calcium ion fluorescent as optoacoustic calcium ion probe;
(2) calcium ion fluorescent is expelled to the region intending detecting;
(3) pulse laser that sends of optoacoustic fluorescence signal excitaton source is after microcobjective, focuses on surveyed area, makes calcium ion fluorescent produce photoacoustic signal and fluorescence signal;
(4) photoacoustic signal is received by bowl-shape hollow focus supersonic detector through after ultrasonic coupling liquid, then amplifies through signal amplifier; Fluorescence signal is received by photomultiplier after spectroscope and optical filter filter; Two paths of signals is gathered by binary channels parallel acquisition card simultaneously, then data is transmitted and be stored into gathering in the computing machine of control software design and image reconstruction software;
(5) detect sample is positioned on three dimensional scanning platform together with the ultrasonic coupling cup holding ultrasonic coupling liquid, is moved by the three-dimensional of the computing machine Quality control that motor control software is housed; Often gathered the photoacoustic signal of a point, three dimensional scanning platform horizontal direction is moved in order and is moved a step; After having scanned a plane, platform has vertically moved a step again; To carry out point by point scanning, for reconstruction of three-dimensional images to sample;
(6) photoacoustic signal of calcium ion fluorescent collected after, reconstruct calcium ion probe light acoustic signal intensity distributed image by maximum value projection algorithm.
8. monitoring method according to claim 7, is characterized in that: also comprise the steps:
(7) by the photoacoustce signal intensity of the calcium ion fluorescent of detection single-point, the change of this calcium ion concentration of Real-Time Monitoring.
9. the monitoring method according to claim 7 or 8, is characterized in that: described calcium ion fluorescent is Rhod-2;
The dominant frequency of described bowl-shape hollow focus supersonic detector is 10MHz, hollow focusing probe;
Maximum value projection algorithm described in step (6) is write voluntarily by Matlab software.
10. monitoring method according to claim 8, is characterized in that:
Detection single-point described in step (7), utilizes the pulsed laser of high repetition frequency 10KHz and the capture card 200M/s of high sampling rate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510896712.0A CN105548102B (en) | 2015-12-07 | 2015-12-07 | Utilize optoacoustic-fluorescence complementary principle deep layer calcium ion concentration monitoring method and device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510896712.0A CN105548102B (en) | 2015-12-07 | 2015-12-07 | Utilize optoacoustic-fluorescence complementary principle deep layer calcium ion concentration monitoring method and device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105548102A true CN105548102A (en) | 2016-05-04 |
| CN105548102B CN105548102B (en) | 2019-06-28 |
Family
ID=55827442
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510896712.0A Expired - Fee Related CN105548102B (en) | 2015-12-07 | 2015-12-07 | Utilize optoacoustic-fluorescence complementary principle deep layer calcium ion concentration monitoring method and device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105548102B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113576415A (en) * | 2021-08-04 | 2021-11-02 | 广东光声医疗科技有限公司 | Four-mode photoacoustic, ultrasonic, fluorescence and optical imaging microscope system |
| CN117030621A (en) * | 2023-09-11 | 2023-11-10 | 深圳大学 | Marking-free laser histological imaging system and imaging method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102621115A (en) * | 2012-02-27 | 2012-08-01 | 华南师范大学 | Confocal simultaneous opto-acoustic imaging and fluorescence imaging method and device |
| CN102928346A (en) * | 2012-10-18 | 2013-02-13 | 中国科学院深圳先进技术研究院 | Dual-module imaging system |
| CN103389273A (en) * | 2013-08-01 | 2013-11-13 | 中国科学院自动化研究所 | Photo-acoustic and optical integrated multi-mode imaging system |
| CN103393408A (en) * | 2013-08-20 | 2013-11-20 | 中国科学院自动化研究所 | Photoacoustic and fluorescent circulation mutual promotion imaging method |
| US20140271475A1 (en) * | 2013-03-12 | 2014-09-18 | The Board Of Trustees Of The Leland Stanford Junior University | Cellulose nanoparticle biodegradable photoacoustic contrast agents |
-
2015
- 2015-12-07 CN CN201510896712.0A patent/CN105548102B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102621115A (en) * | 2012-02-27 | 2012-08-01 | 华南师范大学 | Confocal simultaneous opto-acoustic imaging and fluorescence imaging method and device |
| CN102928346A (en) * | 2012-10-18 | 2013-02-13 | 中国科学院深圳先进技术研究院 | Dual-module imaging system |
| US20140271475A1 (en) * | 2013-03-12 | 2014-09-18 | The Board Of Trustees Of The Leland Stanford Junior University | Cellulose nanoparticle biodegradable photoacoustic contrast agents |
| CN103389273A (en) * | 2013-08-01 | 2013-11-13 | 中国科学院自动化研究所 | Photo-acoustic and optical integrated multi-mode imaging system |
| CN103393408A (en) * | 2013-08-20 | 2013-11-20 | 中国科学院自动化研究所 | Photoacoustic and fluorescent circulation mutual promotion imaging method |
Non-Patent Citations (1)
| Title |
|---|
| 章静波 等: "《细胞生物学实验技术》", 30 June 2006, 化学工业出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113576415A (en) * | 2021-08-04 | 2021-11-02 | 广东光声医疗科技有限公司 | Four-mode photoacoustic, ultrasonic, fluorescence and optical imaging microscope system |
| CN117030621A (en) * | 2023-09-11 | 2023-11-10 | 深圳大学 | Marking-free laser histological imaging system and imaging method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105548102B (en) | 2019-06-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102621115B (en) | Confocal simultaneous opto-acoustic imaging and fluorescence imaging method and device | |
| EP1303756B1 (en) | Apparatus and method for probing light absorbing agents in biological tissues | |
| CN100446730C (en) | Photoacoustic imaging and chromatographic imaging method based on acoustic lens and apparatus thereof | |
| CN105092705B (en) | The multi-modal signal detecting method and device of a kind of steel rail defect | |
| CN107607473B (en) | Simultaneous multipoint excitation and matching received photoacoustic three-dimensional imaging device and method | |
| CN102822661B (en) | Opto-acoustic imaging devices and acousto-optic imaging method | |
| CN103260502B (en) | Subject information acquiring device and subject information acquiring method | |
| CN102944521B (en) | Non-contact photoacoustic and optical coherence tomography dual-imaging device and detection method thereof | |
| CN103389273A (en) | Photo-acoustic and optical integrated multi-mode imaging system | |
| CN102499645A (en) | Photoacoustic and fluorescence dual-mode integrated tomography imaging system and imaging method | |
| CN106124469A (en) | A kind of photoacoustic imaging and optical imagery multi-modal fusion imaging system | |
| CN105595964A (en) | Double-focusing ultrasonic probe and sparse array photoacoustic tomography imaging system | |
| US20160324423A1 (en) | Photoacoustic measurement apparatus and signal processing device and signal processing method for use therein | |
| US20220133273A1 (en) | Transparent ultrasound transducers for photoacoustic imaging | |
| CN1846645A (en) | Photoacoustic blood vessel imaging method and equipment for monitoring photodynamic tumor treating effect | |
| CN101336832A (en) | Pulse type optical acoustic scanning soft-tissue imaging method and device | |
| CN105640493A (en) | Photoacoustic device and control method for photoacoustic device | |
| CN103018171B (en) | Wide-frequency-band optical-acoustic and fluorescent double-imaging device without energy converter and detection method thereof | |
| CN105548102A (en) | Deep calcium ion concentration monitoring method using photoacoustic-fluorescent complementary principle and device using the same | |
| CN104161520A (en) | Epidermal melanin concentration determination method and device based on photoacoustic effect principle | |
| CN105699295B (en) | Utilize the quantitative detecting method and device of optoacoustic fluorescence signal ratio measurement pH value | |
| CN1230125C (en) | Focusing supersonic modulation reflection type optical chromatography imaging method and its apparatus | |
| CN114010152B (en) | Blood brain barrier damage assessment device and method based on dual-wavelength photoacoustic microscopy imaging | |
| Liu et al. | Continuous wave laser excitation based portable optoacoustic imaging system for melanoma detection | |
| CN112535531B (en) | Biological tissue welding effect detection device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190628 Termination date: 20201207 |