CN103728020B - Detect the method for biological ultra-weak photon radiation spectrum - Google Patents

Abstract

The method of detection biological ultra-weak photon radiation spectrum disclosed by the invention, the method is the delayed luminescence curve data of accurate acquisition living body biological sample on different wave length first, then according to the kinetics equation of cell delayed luminescence, least square method is adopted to carry out matching respectively to the measurement data under each wavelength.In the kinetics equation parameter that matching obtains, contain the parameter I describing sample spontaneous luminescence _sL, this parameter is the spontaneous luminescence intensity under each wavelength.By the method, can the spontaneous photon radiation spectrum of indirect inspection sample.Avoid sample spontaneous photon radiation intensity too low, cannot the problem of its spectrum of direct detection.

Description

Detect the method for biological ultra-weak photon radiation spectrum

Technical field

The invention belongs to biological ultraweak photon radiotechnology field, be specifically related to a kind of method detecting biological ultra-weak photon radiation spectrum.

Background technology

Biological ultraweak photon radiation is also called Bio-photon emission, comprise spontaneous luminescence (also referred to as spontaneous photon radiation) and the photoinduced delayed luminescence in the external world (also referred to as photoluminescence), it is a kind of extremely weak quasi-continuous photon radiation of biosystem self radiation, and its luminous intensity is about a hundreds of hv/scm ², wavelength coverage is probably between 180 ~ 800nm.In the various life-informations that crop sends, optical information is the life-information that quantity of information is maximum, the most easily realize nondestructive measurement, so have great importance to its research.

Current is analyze the intensity of luminescence to the research (comprising spontaneous luminescence and delayed luminescence) of biological ultraweak photon radiation mostly.And spectral analysis is as a kind of analytical approach become more meticulous, more life-information can be extracted from the luminescence of biology, a kind of instrument of more strong deciphering life-information is provided, there is important researching value.But, because the ultraweak photon radiation intensity of biology is very faint, adopt the method that the method for grating spectrum or optical filter filtration etc. are traditional, be all difficult to accurately obtain its spectrum.

Summary of the invention

The object of this invention is to provide a kind of method detecting biological ultra-weak photon radiation spectrum, solve because Bio-photon emission intensity is too weak, existing instrument is difficult to the problem measuring its spectrum.

The technical solution adopted in the present invention is, detect the method for biological ultra-weak photon radiation spectrum, adopt a kind of biological ultra-weak photon radiation spectrum detector, its structure is: comprise single-chip microcomputer, single-chip microcomputer is connected to shutter driver module a, LED drive module, radiator valve, shutter driver module b;

Also comprise the photomultiplier, impulse meter, USB interface chip and the computing machine that connect successively;

Also comprise darkroom, the inwall in darkroom is provided with semiconductor chilling plate, darkroom inner bottom part is provided with sample stage, and the side of sample stage is provided with temperature sensor, and the below of sample stage is disposed with shutter b, filter wheel and photomultiplier; The top in darkroom is disposed with shutter a, lens and LED excitation source; LED excitation source is connected with LED drive module; Shutter a is connected with shutter driver module a; Radiator valve is connected with temperature sensor, semiconductor chilling plate respectively; Shutter driver module b is connected with shutter b; Single-chip microcomputer is also connected with USB interface chip;

Specifically implement according to following steps:

Step 1: testing sample is put into darkroom;

Step 2: the wavelength of rotating filtering sheet rotating disk Modifying surveying;

Step 3: the test procedure on start-up simulation machine, arranges light application time and irradiates light intensity, opens LED excitation source irradiate sample by program;

Step 4: after arriving the light application time of previous step setting, program closes LED excitation source automatically, opens shutter simultaneously; Impulse meter starts to add up the sample luminous signal that photomultiplier receives, and luminous for the sample measured in interval photon counting is sent on computing machine and preserves;

Step 5: rotating filtering sheet rotating disk changes measures wavelength, and repeats step 3, step 4 and step 5, until all wavelength measurements needing to measure are complete;

Step 6: the test procedure on computing machine reads above-mentioned test data, simulates the I of the downward tardy light curve of each wavelength _sLparameter, is the spontaneous luminescence spectrum of sample.

Feature of the present invention is also, step 6 wherein is specifically implemented according to following steps:

Adopt least square method, according to formula simulate the I of the downward tardy light curve of each wavelength _sLparameter, wherein I ₀for initial photon number; τ is the characteristic time; β is called attenuation coefficient; I _sLfor the spontaneous luminescence in the unit time; I (t) is the radiation intensity of sample luminescence.

The invention has the beneficial effects as follows, detectable luminous intensity is low to moderate 100hv/scm ²the spectrum of Bio-photon emission, the scope of detecting light spectrum is selected by the combination of different photomultiplier and corresponding optical filter.

Accompanying drawing explanation

Fig. 1 is the biological ultra-weak photon radiation spectrum detector principle assumption diagram that the inventive method adopts;

Fig. 2 is the delayed luminescence curve map of biological sample in different wavelength range in the present invention; Wherein (a) is the delayed luminescence curve of mung bean sprouts under wavelength is respectively 460nm, 475nm, 490nm, 505nm, 520nm; B () is the delayed luminescence curve of mung bean sprouts under wavelength is respectively 535nm, 555nm, 575nm, 590nm, 605nm, 620nm;

Fig. 3 is the Bio-photon emission curve of spectrum detected in the embodiment of the present invention.

In figure, 1.LED excitation source, 2. lens, 3. shutter a, 4. darkroom, 5. semiconductor chilling plate, 6. sample stage, 7. shutter b, 8. filter wheel, 9. photomultiplier, 10. temperature sensor, 11. shutter driver module a, 12.LED driver module, 13. single-chip microcomputers, 14. radiator valves, 15. shutter driver module b, 16. impulse meters, 17.USB interface chip, 18. computing machines.

Embodiment

Below in conjunction with the drawings and specific embodiments, the present invention is described in detail.

Foundation of the present invention is: theoretical according to the coherence of biophoton emission, living cells is the system of a far from equilibrium, the mode that living matter in cell is coupled according to high-order nonlinear associates, define the entirety be highly concerned with, the delayed luminescence of cell life system comes from all interparticle cooperative effects in living matter excited state.The biomolecule i on population being in excited state obeys following nonlinear dynamical equation:

$\frac{\mathrm{dn}}{\mathrm{dt}}=-\mathrm{\μ}\·n^{\mathrm{\γ}}\left(t\right)---\left(1\right)$

The radiation intensity solved by formula (1) is

$I\left(t\right)=\frac{I_0}{{(1+\frac{t}{\mathrm{\τ}})}^{\mathrm{\β}}}---\left(2\right)$

Wherein, μ, γ, β, τ and I ₀be constant, and there is the biological significance determined.I ₀for initial photon number, its size is relevant with the metabolism degree of biosystem; τ is the characteristic time, only relevant with the character of sample self; β is called attenuation coefficient, and it controls the speed of relaxation strongly, and its value can interactional size between each component in characterising biological body.

Consider under external optical excitation, during t → ∞, I (t) should level off to spontaneous luminescence, formula (2) is revised as

$I\left(t\right)=I_{\mathrm{SL}}+\frac{I_0}{{(1+\frac{t}{\mathrm{\τ}})}^{\mathrm{\β}}}---\left(3\right)$

Wherein, I _sLfor the spontaneous luminescence in the unit time.

For the delayed luminescence of an actual biological sample, measured data of experiment and theoretical expression can be done best matching, thus determine parameter beta, τ, I ₀and I _sLvalue.If placing a wavelength between sample and photomultiplier is the optical filter of λ, the spontaneous luminescence intensity I of sample on this wavelength just can be measured _{sL λ}, by converting the optical filter of different wave length, just can obtain the luminous intensity of sample on different wave length, being the spontaneous luminescence spectrum of sample.

The present invention detects the method for biological ultra-weak photon radiation spectrum, irradiates sample by ambient light, after the optical filter filtration of the delayed luminescence specific wavelength produced by sample, then uses the luminous signal after photomultiplier acquisition filter and time domain change procedure thereof.Then parameter I is simulated according to formula (3) _sL, be the relative luminous intensity of sample under current wavelength.Repeat above process, sample relative luminous intensity at multiple wavelengths can be measured, i.e. the luminescent spectrum of sample.

The present invention detects the method for biological ultra-weak photon radiation spectrum, have employed device as shown in Figure 1, comprise LED excitation source 1, lens 2, electronic shutter a3, semiconductor chilling plate 5, electronic shutter b7, filter wheel 8, temperature sensor 10 and photomultiplier 9 and be fixed in the metal shell that is connected with darkroom 4; Sample stage 6 is fixed on bottom darkroom 4, and central authorities have light hole; Single-chip microcomputer 13 connects and controls shutter driver module a11, LED drive module 12, radiator valve 14, USB interface chip 17 and shutter driver module b15; Radiator valve 14 connects semiconductor chilling plate 5; Shutter driver module b15 connects electronic shutter b7; Shutter driver module a11 connects electronic shutter a3; Photomultiplier 9, impulse meter 16, USB interface chip 17 are connected successively with computing machine 18.

LED excitation source 1, for generation of the exciting light irradiating sample.

LED drive module 12, for providing stable working current to LED excitation source 1, and the unlatching of control LED excitation source 1 and extinguishing.

The light that lens 2 make LED send becomes directional light through lens transformation, is radiated on sample uniformly.

Darkroom 4 adopts metallic aluminum material to make, and to reduce the impact of external electromagnetic radiation, avoids the interference of external stray light simultaneously.

Sample stage 6 is used for placing testing sample.

Temperature sensor 10 is used for testing the temperature in darkroom;

Semiconductor chilling plate 5 can to darkroom heating or refrigeration;

The signal that radiator valve 14 is passed back according to preset temperature and temperature sensor 10 controls semiconductor refrigeration chip heating or refrigeration.

Electronic shutter a3 and electronic shutter b7 is used for protecting photomultiplier 9, and when LED excitation source 1 is opened, electronic shutter a3 opens, and electronic shutter b7 closes; After LED excitation source 1 extinguishes, electronic shutter a3 quick closedown, electronic shutter b7 opens, and starts the measurement carrying out delayed luminescence.

Filter wheel 8 is between electronic shutter b7 and photomultiplier 9, and the built-in some optical filters of rotating disk, for being filtered into the monochromatic light of certain wavelength by the Ultra-weak Luminescence of sample.Optical filter can be changed by rotary turnplate, thus selects through wavelength.

Operating voltage when shutter driver module b15 and shutter driver module a11 provides electronic shutter to open and close, shutter driver module exports the voltage of 12V, shutter opening; When shutter driver module output voltage is 0V, shutter close.

Photomultiplier 9 is used for receiving the Ultra-weak Luminescence signal of sample, and light signal is converted to electric pulse, and the luminescence of sample is stronger, then the pulsed frequency exported is higher.

Impulse meter 16 be used for add up photomultiplier 9 export electric impulse signal.

USB interface chip 17(CH341 or PL2303) number of pulses of being added up by impulse meter 16 is sent to computing machine 18 by USB interface.

Run control program and routine analyzer in computing machine 18, before test the parameter of test is sent to single-chip microcomputer by USB interface on the one hand; On the other hand by the pulse of USB interface count pick up, and carry out computational analysis, draw the spectroscopic data of sample luminescence.

Single-chip microcomputer 13 from USB interface chip 17 acceptance test parameter, the peripheral modules such as the workflow of Comprehensive Control instrument and LED drive module 12, radiator valve 14, shutter driver module b15 and shutter driver module a11.

The inventive method is specifically implemented according to following steps:

Step 1: testing sample is put into darkroom 4;

Step 2: the wavelength of rotating filtering sheet rotating disk 8 Modifying surveying;

Step 3: the test procedure on start-up simulation machine 18, arranges light application time and irradiates light intensity, opens LED excitation source 1 irradiate sample by program;

Step 4: after arriving the light application time of previous step setting, program closes LED excitation source 1 automatically, opens shutter 7 simultaneously; Impulse meter 16 starts the sample luminous signal adding up photomultiplier reception, and luminous for the sample measured in interval photon counting is sent to preservation on computing machine 18;

Step 5: rotating filtering sheet rotating disk 8 changes measures wavelength, and repeats step 3, step 4 and step 5, until all wavelength measurements needing to measure are complete;

Step 6: the test procedure on computing machine 18 reads above-mentioned test data, adopts least square method, simulates the I of the downward tardy light curve of each wavelength according to formula (3) _sLparameter, is the spontaneous luminescence spectrum of sample.

Embodiment

Get 20, the sprouting mung bean of 48 hours, be placed in sample cup, put into darkroom, record by above step the delayed luminescence curve that wavelength is 460nm, 475nm, 490nm, 505nm, 520nm, 535nm, 555nm, 575nm, 590nm, 605nm, 620nm, as shown in Figure 2.The curve of spectrum that matching draws is carried out as shown in Figure 3 according to measured result.

Claims (1)

1. detect the method for biological ultra-weak photon radiation spectrum, it is characterized in that, adopt a kind of biological ultra-weak photon radiation spectrum detector, its structure is: comprise single-chip microcomputer (13), single-chip microcomputer (13) is connected to shutter driver module a (11), LED drive module (12), radiator valve (14), shutter driver module b (15);

Also comprise the photomultiplier (9), impulse meter (16), USB interface chip (17) and the computing machine (18) that connect successively;

Also comprise darkroom (4), the inwall in darkroom (4) is provided with semiconductor chilling plate (5), darkroom (4) inner bottom part is provided with sample stage (6), the side of sample stage (6) is provided with temperature sensor (10), and the below of sample stage (6) is disposed with shutter b (7), filter wheel (8) and photomultiplier (9); The top in described darkroom (4) is disposed with shutter a (3), lens (2) and LED excitation source (1); LED excitation source (1) is connected with described LED drive module (12); Shutter a (3) is connected with described shutter driver module a (11); Described radiator valve (14) is connected with temperature sensor (10), semiconductor chilling plate (5) respectively; Described shutter driver module b (15) is connected with shutter b (7); Described single-chip microcomputer (13) is also connected with USB interface chip (17);

Specifically implement according to following steps:

Step 1: testing sample is put into darkroom (4);

Step 2: the wavelength of rotating filtering sheet rotating disk (8) Modifying surveying;

Step 3: the test procedure on start-up simulation machine (18), arranges light application time and irradiates light intensity, opens LED excitation source (1) irradiate sample by program;

Step 4: after arriving the light application time of previous step setting, program closes LED excitation source (1) automatically, opens shutter b (7) simultaneously; Impulse meter (16) starts the sample luminous signal adding up photomultiplier reception, and luminous for the sample measured in interval photon counting is sent to the upper preservation of computing machine (18);

Step 5: rotating filtering sheet rotating disk (8) changes measures wavelength, and repeats step 3, step 4 and step 5, until all wavelength measurements needing to measure are complete;

Step 6: the test procedure on computing machine (18) reads above-mentioned test data, simulates the I of the downward tardy light curve of each wavelength _sLparameter, is the spontaneous luminescence spectrum of sample, specifically implements according to following steps:

Adopt least square method, according to formula simulate the I of the downward tardy light curve of each wavelength _sLparameter, wherein I ₀for initial photon number; τ is the characteristic time; β is called attenuation coefficient; I _sLfor the spontaneous luminescence in the unit time; I (t) is the radiation intensity of sample luminescence.