CN107589073A - A kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method and device - Google Patents
A kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method and device Download PDFInfo
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- CN107589073A CN107589073A CN201710773584.XA CN201710773584A CN107589073A CN 107589073 A CN107589073 A CN 107589073A CN 201710773584 A CN201710773584 A CN 201710773584A CN 107589073 A CN107589073 A CN 107589073A
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Abstract
The invention discloses a kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method and device, described device mainly includes LASER Light Source, deuterium lamp, the first moving stop, the second moving stop, plane mirror, convex lens, spectrometer and PC；Wherein the first moving stop is located between the LASER Light Source and plane mirror, available for blocking the LASER Light Source, after the second moving stop is located at the deuterium lamp, available for blocking deuterium lamp light source；The plane mirror is used to reflex to LASER Light Source in cuvette, the convex lens are exposed in cuvette for the diverging light of deuterium lamp transmitting to be changed into directional light, transmitted light after the deuterium lamp irradiation cuvette is by fiber optic conduction into spectrometer, and the spectrometer is by data transfer to PC.Have the advantages that signal to noise ratio is high, accuracy of measurement is high, measurement efficiency is high and automatization level is high using the sensitising agent singlet oxygen quantum yield measuring method and device of the present invention.
Description
Technical field
The present invention relates to a kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method and device, belong to light power
Treatment technology field.
Background technology
Optical dynamic therapy (Photodynamic therapy, PDT) is very long for treating and diagnosing shallow cancer
Time.Its treat principle be：After sensitising agent is injected into human organ, sensitising agent is selectively absorbed and is gathered in
Near tumour cell；Light irradiation pathological tissues are excited using suitable wavelength, sensitising agent is excited production in the presence of exciting light
Raw singlet oxygen；Singlet oxygen selectively kills tumour cell, so as to reach the effect for the treatment of.
The effect of optical dynamic therapy is main and oxygen molecule concentration, photosensitizer concentration, excitating light strength and singlet oxygen (1O2) measure
Sub- yield (ΦΔ) relevant.Because the effect of optical dynamic therapy, depends on the cytotoxicity of singlet oxygen, therefore, singlet oxygen amount
Sub- yield is the significant effects factor of optical dynamic therapy curative effect.And in fact, the singlet oxygen quantum yield of sensitising agent easily by
The influence of environmental factor and storage time.Therefore, it is necessary the singlet oxygen amount of accurate measurement sensitising agent before optical dynamic therapy
Sub- yield.And the singlet oxygen quantum yield of direct measurement sensitising agent is extremely difficult.The medicines such as DPBF and NADH can and singlet oxygen
Reaction, its change in concentration can react singlet oxygen yield i.e. singlet oxygen quantum yield very well.Therefore, the medicine such as DPBF and NADH can
Singlet oxygen quantum yield to be used for measuring sensitising agent as singlet oxygen indicator.Compared with other singlet oxygen indicators, DPBF
Have the advantages that absorption spectra is more preferable and singlet oxygen reaction faster, it is sensitiveer, accurate.
Therefore, patent of the present invention designs a kind of photosensitizer for photodynamic therapy singlet using DPBF as singlet oxygen indicator
Oxygen quantum yield measuring method and device.
The content of the invention
For overcome the deficiencies in the prior art, it is an object of the invention to provide a kind of photosensitizer for photodynamic therapy singlet oxygen amount
Sub- yield measuring method and device.
To achieve the above object, the present invention adopts the following technical scheme that：
A kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method, comprises the following steps：
(1) spectrometer is started, cuvette is sky, blocks laser and deuterium lamp, back end spectrum is measured with spectrometer；
(2) mixed solution of sensitising agent to be measured and methanol is put into cuvette, blocks laser and open deuterium lamp, surveyed with spectrometer
Obtain the mixed liquor of sensitising agent and methanol passes through spectrum I ' (λ), as calibration spectrum；
Wherein I ' (λ)=I0(λ)e-σ′(λ)N′L, I0(λ) is the spectrum that passes through of methanol, and σ ' (λ) is the absorption of sensitising agent to be measured
Sectional area, N' are the concentration of sensitising agent to be measured, and L is optical path length；
(3) sensitising agent to be measured, methanol and DPBF mixed solution are put into cuvette；
(4) block laser and open deuterium lamp, with spectrometer measure sensitising agent to be measured, methanol and DPBF through spectrum I (λ)；
Wherein I (λ)=I0(λ)e-σ(λ)NLe-σ′(λ)N′L, σ (λ) is DPBF absorption cross section, and N is DPBF concentration；
(5) extractThe crest of function, wherein functionWherein k=σ (λ) L；
(6) open laser and block deuterium lamp, laser is irradiated cuvette, sensitising agent to be measured produces singlet oxygen, the singlet oxygen with
Oxidation reaction occurs for DPBF；
(7) system automatic decision has completed the pendulous frequency whether pendulous frequency is less than default, if "Yes", returns
Step (4), if "No", carry out step (8)；
(8) the multiple crests for extracting step (5) are ordinate, and corresponding laser irradiation time is abscissa, using most
Small square law is fitted, and the slope of matched curve is the k of sensitising agent to be measured；
(9) open laser and block deuterium lamp, cuvette is measured the emission spectrum of exciting light, normalization with spectrometer for sky
Obtain spectrum I " (λ)；
(10) open laser and block deuterium lamp, methanol is put into cuvette, the transmission spectrum I of methanol is measured with spectrometer0
(λ), utilize I'(λ) divided by I0(λ), obtains e-σ′(λ)N′L, ask for integration according to equation below and obtain the absorption light of sensitising agent to be measured
Compose Iabs, Iabs=∫ I " (λ) × (1-e-σ′(λ)N′L)dλ；
(11) above method is used under equal experiment condition, measures the k of the rose-red as reference photosensitive agentrefWith
The singlet oxygen quantum yield of rose-red is obtained by existing document
(12) byCalculate the singlet oxygen quantum yield φ of tested sensitising agentΔ。
Preferably, the equipment blocked laser or blocked used in deuterium lamp is the steering wheel being controlled by the controller and connected with the steering wheel
The moving stop connect.
Preferably, in the mixed liquor of the sensitising agent to be measured and methanol, methanol is solvent, and sensitising agent to be measured is solute.Treat
Its concentration is 2 μM when survey sensitising agent is hematoporphyrin monomethyl ether (HMME), and sensitising agent to be measured is aloe-emodin (Aloe-emodin)
When its concentration be 50 μM；Concentration as the rose-red (Rose Bengal) of reference photosensitive agent is 1 μM, the sensitising agent to be measured,
DPBF concentration is 30 μM in methanol and DPBF mixed liquor.
Preferably, spectrometer by optical fiber data measured and transfers data to PC.
Preferably, built with the upper computer software based on LabVIEW, the parameter for completing spectrometer is set the PC
Put, store and handle the measurement data of spectrometer and the calculating of singlet oxygen quantum yield.
Preferably, in step (4), extractionCrest using LabVIEW functions extract.
Present invention also offers a kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measurement apparatus, mainly including laser
Light source, deuterium lamp, the first moving stop, the second moving stop, plane mirror, convex lens, spectrometer and PC；
First moving stop is located between the LASER Light Source and plane mirror, available for blocking the laser light
Source, after second moving stop is located at the deuterium lamp, available for blocking deuterium lamp light source；
For the laser reflection that the plane mirror is used to send LASER Light Source into cuvette, the convex lens are used for will
The diverging light of deuterium lamp transmitting changes into directional light and exposed in cuvette, and the transmitted light after the deuterium lamp irradiation cuvette passes through optical fiber
Into spectrometer, the spectrometer is used for measurement data and is transmitted to PC for conduction.
Preferably, first moving stop and second moving stop are connected with the first steering wheel, the second steering wheel respectively,
First steering wheel and the second steering wheel are connected by controller with PC, and PC sends instruction, and steering wheel is controlled by controller
Movement that is mobile and then controlling moving stop.
Relative to prior art, the beneficial effects of the present invention are：
With singlet oxygen cold light dosimetry of the prior art, singlet oxygen recessiveness dosimetry and the clear and definite dosimetry phase of singlet oxygen
Than, sensitising agent singlet oxygen quantum yield measuring method of the invention uses sensitising agent and singlet oxygen indicator DPBF full spectrum,
With higher signal to noise ratio；Using DPBF as singlet oxygen indicator, compared with other singlet oxygen indicators such as NADH and APDA,
DPBF have the advantages that more preferable absorption spectra, it is more sensitive to singlet oxygen and with singlet oxygen react it is more rapid；Using tested
Method of the absorption spectrum of sensitising agent and DPBF mixed solutions compared with the absorption spectrum of tested sensitising agent, obtains DPBF's
Absorption spectrum, so as to try to achieve DPBF attenuation rate i.e. matched curve slope k, there is higher measurement accuracy；Measurement starts with
Spectrometer measures back end spectrum, subtracts back end spectrum through spectrum by that will measure in follow-up measurement process, can eliminate
Influence of the environment to measurement；The measuring system of sensitising agent singlet oxygen quantum yield based on the present invention can be automatically performed singlet oxygen amount
The measurement of sub- yield, there is more high measurement efficiency and automatization level；Using controller, steering wheel, moving stop realize deuterium lamp and
The automatic switchover of laser, it can avoid frequently switching on deuterium lamp and laser, extend the service life of deuterium lamp and laser.
Brief description of the drawings
Fig. 1 is the flow chart of the measuring method of the present invention.
Fig. 2 is the structure chart of the measurement apparatus of the present invention.
Description of reference numerals：1- LASER Light Sources；2- deuterium lamps；The moving stops of 3- first；The moving stops of 4- second；5- planes are anti-
Penetrate mirror；6- convex lens；7- spectrometers；8-PC machines；9- cuvettes；The steering wheels of 10- first；The steering wheels of 11- second；12- controllers；13-
Optical fiber.
Embodiment
In conjunction with embodiment, the present invention is described in more detail.It should be appreciated that the present embodiment is merely to illustrate this hair
It is bright, rather than limit protection scope of the present invention.
A kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method, comprises the following steps：
(1) start spectrometer 7, open indicator lamp, cuvette 9 is empty, control of first moving stop 3 in the first steering wheel 10
Under block LASER Light Source 1, the second moving stop 4 blocks deuterium lamp 2 under the control of the second steering wheel 11, and back end is measured with spectrometer 7
Spectrum；
(2) by the mixed solution of sensitising agent to be measured and methanol, (now sensitising agent to be measured is hematoporphyrin monomethyl ether (HMME), blood
The concentration of porphyrin monomethyl ether (HMME) is 2 μM), cuvette is put into, LASER Light Source 1 is blocked and opens deuterium lamp 2, measured with spectrometer 7
The mixed liquor of hematoporphyrin monomethyl ether and methanol passes through spectrum I ' (λ)；
Wherein I ' (λ)=I0(λ)e-σ′(λ)N′L, I0(λ) is the spectrum that passes through of methanol, and σ ' (λ) is the absorption of sensitising agent to be measured
Sectional area, N' are the concentration of sensitising agent to be measured, and initial concentration is 2 μM, and L is optical path length, specially 1cm；
(3) sensitising agent to be measured, methanol and DPBF mixed solution are put into (the wherein hematoporphyrin monomethyl ether of cuvette 9
(HMME) initial concentration is 2 μM, and DPBF initial concentration is 30 μM)；
(4) block LASER Light Source 1 and open deuterium lamp 2, sensitising agent, methanol and DPBF mixed solutions to be measured are measured with spectrometer 7
Through spectrum I (λ), as calibration spectrum；
Wherein I (λ)=I0(λ)e-σ(λ)NLe-σ′(λ)N′L, σ (λ) is DPBF absorption cross section, and N is DPBF concentration, initially
Concentration is 30 μM；
(5) extracted using LabVIEW functionsThe crest of function, wherein k=σ (λ) L, σ (λ), N
For variable；
(6) open LASER Light Source 1 and block deuterium lamp 2, laser is irradiated cuvette 9, sensitising agent to be measured produces singlet oxygen, the list
With DPBF oxidation reaction occurs for state oxygen；
(7) system automatic decision has completed measurement 8-12 times whether pendulous frequency is less than default, if "Yes", returns
Step (4) is returned, if "No", carries out step (8)；
(8) the multiple crests for extracting step (5) are ordinate, and corresponding laser irradiation time is abscissa, using most
Small square law is fitted, and the slope of matched curve is the k of sensitising agent to be measured, and sensitising agent to be measured is hematoporphyrin monomethyl ether (HMME)
When its k be 0.028；
(9) open LASER Light Source 1 and block deuterium lamp 2, cuvette 9 is measured the transmitting light of exciting light with spectrometer 7 for sky
Spectrum, normalization obtain spectrum I " (λ)；
(10) open LASER Light Source 1 and block deuterium lamp 2, methanol is put into cuvette 9, the transmission of methanol is measured with spectrometer 7
Spectrum I0(λ), utilize I'(λ) divided by I0(λ), obtains e-σ′(λ)N′L, ask for integration according to equation below and obtain sensitising agent to be measured
Absorption spectrum Iabs, Iabs=∫ I " (λ) × (1-e-σ′(λ)N′L) d λ, its I when sensitising agent to be measured is hematoporphyrin monomethyl ether (HMME)abs
For 0.505；
(11) above method is used under equal experiment condition, measures the k of the rose-red as reference photosensitive agentrefWith
The singlet oxygen quantum yield of rose-red is obtained by existing documentUse laser excitation sensitising agent of the wavelength for 532nm
Corresponding k during rose-redref=0.046,
(12) byCalculate the singlet oxygen quantum yield φ of hematoporphyrin monomethyl ether (HMME)Δ=0.77.
Present invention also offers a kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measurement apparatus, including LASER Light Source
1st, deuterium lamp 2, the first moving stop 3, the second moving stop 4, plane mirror 5, convex lens 6, spectrometer 7, PC 8,9- colorimetrics
Ware, the steering wheels of 10- first, the steering wheels of 11- second, 12- controllers and 13- optical fiber.Wherein, the first moving stop 3 is located at the laser
Between light source 1 and plane mirror 5, available for the LASER Light Source 1 is blocked, second moving stop 4 is located at the deuterium lamp 2
Afterwards, available for blocking deuterium lamp light source, and the first moving stop 3 is connected with the first steering wheel 10, the second moving stop 4 and the second steering wheel
11 connections, the steering wheel 11 of the first steering wheel 10 and second are connected by controller 12 with PC 8, and PC 8 sends instruction, passes through control
Device 12 controls the movement of the first steering wheel 10 and the second steering wheel 11 and then controls the shifting of the first moving stop 3 and the second moving stop 4
It is dynamic.
Plane mirror 5 is used for by the laser reflection that LASER Light Source 1 is sent into cuvette 9, and the convex lens 6 are used for will
The diverging light that deuterium lamp 2 is launched changes into directional light and exposed in cuvette 9, and the transmitted light after the irradiation of deuterium lamp 2 cuvette 9 passes through
Optical fiber 13 is conducted into spectrometer 7, and the spectrometer 7 is by optical fiber measurement data and is transmitted to PC 8, in the PC 8
Equipped with the upper computer software based on LabVIEW, for completing parameter setting, storage and the measurement for handling spectrometer 7 of spectrometer 7
Data and the calculating for calculating singlet oxygen quantum yield.
Embodiments of the invention content is disclosed above, but the present embodiment is not limited to the scope that the present invention is implemented,
The simple equivalent changes and modifications made according to claims of the present invention and description, still falls within the technology of the present invention
In the range of scheme.
Claims (8)
1. a kind of photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method, it is characterised in that comprise the following steps：
(1) spectrometer is started, cuvette is sky, blocks laser and deuterium lamp, back end spectrum is measured with spectrometer；
(2) mixed solution of sensitising agent to be measured and methanol is put into cuvette, blocks laser and open deuterium lamp, light is measured with spectrometer
The mixed liquor of quick dose and methanol through spectrum I ' (λ), as calibration spectrum；
Wherein I ' (λ)=I0(λ)e-σ′(λ)N′L, I0(λ) is the spectrum that passes through of methanol, and σ ' (λ) is the absorption cross-section of sensitising agent to be measured
Product, N' are the concentration of sensitising agent to be measured, and L is optical path length；
(3) sensitising agent to be measured, methanol and DPBF mixed solution are put into cuvette；
(4) block laser and open deuterium lamp, with spectrometer measure sensitising agent to be measured, methanol and DPBF through spectrum I (λ)；
Wherein I (λ)=I0(λ)e-σ(λ)NLe-σ′(λ)N′L, σ (λ) is DPBF absorption cross section, and N is DPBF concentration；
(5) extractThe crest of function, wherein functionWherein k=σ (λ) L；
(6) open laser and block deuterium lamp, laser is irradiated cuvette, sensitising agent to be measured produces singlet oxygen, the singlet oxygen and DPBF
Generation oxidation reaction；
(7) system automatic decision has completed the pendulous frequency whether pendulous frequency is less than default, if "Yes", return to step
(4), if "No", step (8) is carried out；
(8) the multiple crests for extracting step (5) are ordinate, and corresponding laser irradiation time is abscissa, using a most young waiter in a wineshop or an inn
Multiplication is fitted, and the slope of matched curve is the k of sensitising agent to be measured, namely DPBF attenuation rate；
(9) open laser and block deuterium lamp, cuvette is measured the emission spectrum of exciting light with spectrometer for sky, normalization obtains
Spectrum I " (λ)；
(10) open laser and block deuterium lamp, methanol is put into cuvette, the transmission spectrum I of methanol is measured with spectrometer0(λ), utilize
I'(λ) divided by I0(λ), obtains e-σ′(λ)N′L, ask for integration according to equation below and obtain the absorption spectrum I of sensitising agent to be measuredabs,
Iabs=∫ I " (λ) × (1-e-σ′(λ)N′L)dλ；
(11) above method is used under equal experiment condition, measures the k of the rose-red as reference photosensitive agentrefWithBy existing
Document obtains the singlet oxygen quantum yield of rose-red
(12) byCalculate the singlet oxygen quantum yield φ of tested sensitising agentΔ。
2. photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method as claimed in claim 1, it is characterised in that block
Laser or the equipment blocked used in deuterium lamp are the steering wheel being controlled by the controller and the moving stop being connected with the steering wheel.
3. photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method as claimed in claim 1, it is characterised in that described
Methanol is solvent in the mixed liquor of sensitising agent and methanol to be measured, and sensitising agent is solute, and sensitising agent to be measured is hematoporphyrin monomethyl ether
(HMME) its concentration is 2 μM when, and its concentration is 50 μM when sensitising agent to be measured is aloe-emodin (Aloe-emodin), as ginseng
The concentration for examining the rose-red (Rose Bengal) of sensitising agent is 1 μM；
DPBF concentration is 30 μM in the mixed liquor of the sensitising agent to be measured, methanol and DPBF.
4. photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method as claimed in claim 1, it is characterised in that spectrum
Instrument is by optical fiber data measured and transfers data to PC.
5. photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method as claimed in claim 4, it is characterised in that described
PC is built with the upper computer software based on LabVIEW, for completing the parameter setting of spectrometer, storage and handling spectrometer
The calculating of measurement data and singlet oxygen quantum yield.
6. photosensitizer for photodynamic therapy singlet oxygen quantum yield measuring method as claimed in claim 1, it is characterised in that step
(4) in, extractionCrest using LabVIEW functions extract.
A kind of 7. photosensitizer for photodynamic therapy singlet oxygen quantum yield measurement apparatus, it is characterised in that mainly including LASER Light Source,
Deuterium lamp, the first moving stop, the second moving stop, plane mirror, convex lens, spectrometer and PC；
First moving stop is located between the LASER Light Source and plane mirror, available for blocking the LASER Light Source,
After second moving stop is located at the deuterium lamp, available for blocking deuterium lamp light source；
Into cuvette, the convex lens are used for deuterium lamp the laser reflection that the plane mirror is used to send LASER Light Source
The diverging light of transmitting changes into directional light and exposed in cuvette, and the transmitted light after the deuterium lamp irradiation cuvette passes through fiber optic conduction
Into spectrometer, the spectrometer is used for measurement data and is transmitted to PC.
8. photosensitizer for photodynamic therapy singlet oxygen quantum yield measurement apparatus as claimed in claim 7, it is characterised in that described
First moving stop and second moving stop are connected with the first steering wheel, the second steering wheel respectively, first steering wheel and second
Steering wheel is connected by controller with PC, and PC sends instruction, and the movement of steering wheel is controlled and then the mobile gear of control by controller
The movement of plate.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111693500A (en) * | 2020-06-19 | 2020-09-22 | 哈尔滨工业大学 | Method for realizing monitoring of singlet oxygen quantum yield based on time-resolved spectral measurement |
-
2017
- 2017-08-31 CN CN201710773584.XA patent/CN107589073A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| PENG WANG等: "Luminescence and photosensitivity of gadolinium labeled hematoporphyrin monomethyl ether", 《OPTICS EXPRESS》 * |
| WOLFGANG SPILLER等: "Singlet Oxygen Quantum Yields of Different Photosensitizers in Polar Solvents and Micellar Solutions", 《JOURNAL OF PORPHYRINS AND PHTHALOCYANINES》 * |
| 王鹏: "光动力反应中单态氧剂量分布的研究", 《中国博士学位论文全文数据库医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111693500A (en) * | 2020-06-19 | 2020-09-22 | 哈尔滨工业大学 | Method for realizing monitoring of singlet oxygen quantum yield based on time-resolved spectral measurement |
| CN111693500B (en) * | 2020-06-19 | 2022-11-25 | 哈尔滨工业大学 | Method for realizing monitoring of singlet oxygen quantum yield based on time-resolved spectral measurement |
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