CN206618657U - A kind of device for studying molecular migration motion - Google Patents
A kind of device for studying molecular migration motion Download PDFInfo
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- CN206618657U CN206618657U CN201720454499.2U CN201720454499U CN206618657U CN 206618657 U CN206618657 U CN 206618657U CN 201720454499 U CN201720454499 U CN 201720454499U CN 206618657 U CN206618657 U CN 206618657U
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Abstract
The utility model is related to optical microphotograph field, a kind of device for studying molecular migration motion, laser, the polarizer, Pockers cell, analyzer, optical splitter, sample is collinear, the laser beam of the laser transmitting is enabled to pass through the polarizer, Pockers cell, two beam laser are divided into by the optical splitter after analyzer, beam of laser is passed through apart from L direct irradiations in sample surfaces, another beam of laser reaches level crossing after apart from a, sample surfaces are irradiated in after level crossing reflects, the above-mentioned two beam laser for being irradiated in the sample surfaces are angle o, the interference figure of light can be produced at sample surfaces, it is described adjustable apart from a and L, so as to change sample surfaces interference fringe spacing i by the value for changing the angle θ；The level crossing is connected with displacement platform and position modulation can be carried out to the level crossing so that the level crossing carries out sinusoidal vibration along its normal orientation with frequency 800Hz, so as to change position of interference fringe at the sample surfaces.
Description
Technical field
The utility model is related to optical microphotograph field, particularly a kind of analysis it is more simple and it is accurate, expansion can be measured
Dissipate a kind of device of research molecular migration motion of coefficient.
Background technology
Optically powered is also known as photobleaching, refers to fluorescent material is ejected under the irradiation of light fluorescence intensity over time
The phenomenon that passage progressively weakens or even disappeared.The quality of fluorescence imaging is largely dependent upon fluorescence signal intensity, improves and swashs
Luminous intensity can improve signal intensity, but when the intensity of exciting light exceedes certain limit, light absorbs tend to saturation, not
Excited state molecule is reversibly destroyed, here it is photobleaching phenomenon.Fluorescence recovery after photobleaching is to use lipophilicity or hydrophily
Fluorescence molecule, such as fluorescein, green fluorescent protein and albumen or lipid couple, and for detecting, mark molecule is thin in live body
Cellular surface or cell interior motion and its migration rate.The principle of recovery technology after fluorescent bleach is：Shone using superlaser
The a certain specific region of cell is penetrated, makes the fluorescence molecule of the region internal labeling is irreversible to be quenched, this region claims fluorescent bleach
Area.Subsequently, as the motion of the lipid molecular or protein molecule in cytoplasm, surrounding non-bleached area fluorescence molecule is constantly to light
Bleach area migration, the fluorescence intensity in fluorescent bleach area is little by little returned to previous level, on this basis in the urgent need to
It is a kind of that there is the device and method that can be used in measuring diffusion coefficient.
What the method that fluorescence recovers after traditional use photobleaching had to study the technology of the motions such as molecular migration diffusion
Defect has：In order to reach the demand of light signal collection in experiment, the density needs of fluorescence molecule are very high；The measurement of beam profile is needed
It is very accurate；The monitoring of photobleaching process is not readily reachable by, so as to need higher light intensity to carry out photobleaching, is so possible to
The structure of sample molecule can be destroyed, particularly some biological sample molecules.A kind of device for studying molecular migration motion makes
Photobleaching is carried out to sample with the hot spot of plane and subsequent monitoring detection, solve drawbacks described above.
Utility model content
In order to solve the above problems, the utility model device causes the light that light and dark striated is produced in sample surfaces
Spot region, after carrying out photobleaching with strong light in bright fringes region, continues to keep relatively low intensity illumination, to detect between light and shade striped
Contrast change with time, and obtain by subsequent analysis the information such as the diffusion coefficient of testing molecule, Blang's diffusion system
Number can be obtained independently, and it is the average value in whole spot pattern region.
The technical scheme that the utility model is used is：
It is described it is a kind of study molecular migration motion device mainly include laser, the polarizer, Pockers cell, analyzer,
Optical splitter, sample, displacement platform, level crossing, optical fiber, optical filter, photomultiplier, lock-in amplifier, cable, the lock mutually amplify
Device connects the Pockers cell and the displacement platform by cable respectively, the luminous energy of the sample surfaces by the optical fiber and
The optical filter is transferred in the photomultiplier, and the Pockers cell can make laser generation one extremely short less than 1 second
Full intensity pulse, the photomultiplier, which can collect after photobleaching the emerging fluorescence due to molecule diffusion motion, to be believed
Number, then analyzed by the lock-in amplifier, so as to obtain the sample surfaces bright fringes part and dark fringe part
Fluorescence intensity average contrast C (t).
The laser, the polarizer, Pockers cell, analyzer, optical splitter, sample are collinear, enable to the laser
The laser beam of transmitting is divided into two beam laser, beam of laser after the polarizer, Pockers cell, analyzer by the optical splitter
By apart from L direct irradiations, in the sample surfaces, another beam of laser reaches the level crossing after apart from a, by described
The sample surfaces are irradiated in after level crossing reflection, the above-mentioned two beam laser for being irradiated in the sample surfaces are angle o, can be in institute
The interference figure that light is produced at sample surfaces is stated, it is described adjustable apart from a and L, so as to be changed by the value for changing the angle θ
The sample surfaces interference fringe spacing i；The level crossing is connected with the displacement platform and the level crossing can be carried out
Position modulation so that the level crossing carries out sinusoidal vibration along its normal orientation with frequency 800Hz, so as to change the sample table
Position of interference fringe at face.
Use the method and step that the utility model device is studied for：
The laser beam that laser described in one, is launched is after the polarizer, Pockers cell, analyzer, by the light splitting
Device is divided into two beam laser, and beam of laser is passed through apart from L direct irradiations in the sample surfaces, and another beam of laser is after apart from a
The level crossing is reached, the sample surfaces are irradiated in after level crossing reflection, it is above-mentioned to be irradiated in the sample surfaces
Two beam laser it is angle o so that at the sample surfaces produce light interference figure；
Two, can change the value of the angle θ by adjusting the length apart from a and L, so as to change the sample table
Face interference fringe spacing i；
Three, make the pulse of the laser one extremely short full intensity less than 1 second of generation by the Pockers cell, to bright fringes
The molecule of the corresponding species of the fluorescence labeling at place carries out optically powered, and after optically powered process terminates, the laser is continuously launched
Light intensity is I (r, t) laser to detect the change in fluorescence of the sample surfaces；
Four, carry out position modulation by the displacement platform to the level crossing so that the level crossing is along its normal orientation
Sinusoidal vibration is carried out with frequency 800Hz, so as to change position of interference fringe at the sample surfaces；
Described in five, photomultiplier collect photobleaching after due to molecule diffusion motion emerging fluorescence signal, then
Analyzed by the lock-in amplifier, thus obtain sample surfaces bright fringes part and dark fringe part fluorescence intensity it
Than i.e. average contrast C (t), fluorescence signal F (t) and the concentration for the molecule being fluorescently labeled that the photomultiplier is measured
cm(r, t) and detection light intensity I (r, t) are relatedWhereinWithIt is cm(r, t) and I (r, t) spatial Fourier transform, F (t) can be decomposed into the humorous of modulating frequency
Wave scale number, in experiment, the harmonic component f of single order and second order1And f (t)2(t) can simultaneously it be obtained by the lock-in amplifier；
Experimental data final six, provides the average contrast between dark fringe and bright fringes
Above formula characterizes Blang's dispersal behavior with an exponential damping；
Seven,Relaxation time τ can be obtainedq, by relaxation time τqWith the relation of diffusion coefficient DQ is the space periodic of candy strip, can obtain diffusion coefficient D.
The sample surfaces can add a photomask for having circular open, and the measured zone of the sample surfaces is located at institute
State in opening, the material of photomask is to the reflectivity very little of light, can reduce due to the sample surfaces that laser facula is caused
Influence of the veiling glare of non-measured region to experiment signal to noise ratio；Different optical maser wavelength 488nm, 532nm, 633nm can be used
Etc. being tested, and the result that each optical maser wavelength condition experiment of Integrated comparative is obtained, can more accurately reflect molecule
Travel motion；To large biological molecule sample, laser intensity is less than 250W/cm during photobleaching2, and the burst length is less than 400ms.
Pockers cell is a kind of electro-optical device, and it includes an electro-optic crystal passed through by light, based on Pockels effect,
The polarization direction of light can be regulated and controled by the voltage being applied on crystal and change light and pass through the bit phase delay after crystal.
Due to the diffusion of molecule, the contrast C (t) reduces with the time, using same candy strip come to sample
Product carry out optically powered and detection and collect signal ensure that them by same wavelength to characterize.Visited in the lock-in amplifier
The composition of the fundamental frequency of the fluorescence intensity of survey is changed over time, and meets the diffusion equation of single mode, and contrast C (t) is according to simple
Exponential dampingWherein relaxation time τqIt is relevant with diffusion coefficient DQ is bar graph
The space periodic of case, so, by changing q, can verify that Blang's diffusion rule and accurately determines the diffusion system of brown-Zsigmondy movements
Number.Constant C0、C1With brown-Zsigmondy movements and not relative fractions between brown-Zsigmondy movementsWithIt is relevant.
In the situation of the complex sample containing N kind different motion populations,It can be extended to
The diffusion coefficient and fraction of kth kind population be respectively
The beneficial effects of the utility model are：
The utility model analysis is more simple and accurate, it is easier to detects the small change of diffusion coefficient, or differentiates not
With the difference between population；Carrying out optically powered and detection to sample and collect the periodic sinusoidal pattern used in signal can be with
Diffusion equation for detecting a single mode, the pattern of optically powered can be an increasingly complex figure, usually Gauss light
Spot；In addition, the utility model device can more easily change the parameter of interference fringe, to be spread in reciprocal space Direct Test
The authenticity of rule, the device read and the same pattern of optically powered, can be applied to polymer the diffusion at interface research,
The field such as DNA diffusions and electrophoresis.
Brief description of the drawings
Further illustrated with reference to figure of the present utility model：
Fig. 1 is the utility model schematic diagram.
In figure, 1. lasers, 2. polarizers, 3. Pockers cells, 4. analyzers, 5. optical splitters, 6. samples, 7. displacement platforms,
8. level crossing, 9. optical fiber, 10. optical filters, 11. photomultipliers, 12. lock-in amplifiers, 13. cables.
Embodiment
If Fig. 1 is the utility model schematic diagram, a kind of device for studying molecular migration motion mainly includes laser
1st, the polarizer 2, Pockers cell 3, analyzer 4, optical splitter 5, sample 6, displacement platform 7, level crossing 8, optical fiber 9, optical filter 10, photoelectricity
Multiplier tube 11, lock-in amplifier 12, cable 13, the lock-in amplifier 12 connect the Pockers cell 3 respectively by cable 13
With the displacement platform 7, the luminous energy on the surface of sample 6 is transferred to the photoelectricity by the optical fiber 9 and the optical filter 10
In multiplier tube 11, the Pockers cell 3 can make the pulse of the laser one extremely short full intensity less than 1 second of generation, the light
Electric multiplier tube 11 can collect after photobleaching the emerging fluorescence signal due to molecule diffusion motion, then mutually be put by the lock
Big device 12 is analyzed, so as to obtain average pair of the surface bright fringes part of sample 6 and the fluorescence intensity of dark fringe part
Than degree C (t).
The laser 1, the polarizer 2, Pockers cell 3, analyzer 4, optical splitter 5, sample 6 is collinear, enables to described
The laser beam that laser 1 is launched is that two beams swash by 5 points of the optical splitter after the polarizer 2, Pockers cell 3, analyzer 4
Light, beam of laser is passed through apart from L direct irradiations in the surface of sample 6, and another beam of laser reaches the plane after apart from a
Mirror 8, is irradiated in the surface of sample 6 after the level crossing 8 reflection, and above-mentioned two beams for being irradiated in the surface of sample 6 swash
Light is angle o, and the interference figure of light can be produced at the surface of sample 6, described adjustable apart from a and L, so as to by changing
The value for becoming the angle θ changes the surface fringe spacing i of sample 6；Be connected with the displacement platform 7 level crossing 8 and
Position modulation can be carried out to the level crossing 8 so that the level crossing 8 carries out sinusoidal along its normal orientation with frequency 800Hz
Vibration, so as to change position of interference fringe at the surface of sample 6.
Claims (1)
1. a kind of device for studying molecular migration motion, mainly including laser (1), the polarizer (2), Pockers cell (3), analyzing
Device (4), optical splitter (5), sample (6), displacement platform (7), level crossing (8), optical fiber (9), optical filter (10), photomultiplier
(11), lock-in amplifier (12), cable (13), the lock-in amplifier (12) connect the Pu Keer respectively by cable (13)
Box (3) and the displacement platform (7), the luminous energy on sample (6) surface are passed by the optical fiber (9) and the optical filter (10)
Defeated to arrive in the photomultiplier (11), the Pockers cell (3) can make laser produce one and be less than 1 second extremely short full strong
The pulse of degree, the photomultiplier (11) can collect after photobleaching the emerging fluorescence signal due to molecule diffusion motion,
Then analyzed by the lock-in amplifier (12), so as to obtain the sample (6) surface bright fringes part and dark fringe
The average contrast C (t) of partial fluorescence intensity,
It is characterized in that：The laser (1), the polarizer (2), Pockers cell (3), analyzer (4), optical splitter (5), sample (6)
Collinear, the laser beam for enabling to the laser (1) to launch passes through the polarizer (2), Pockers cell (3), analyzer (4)
Two beam laser are divided into by the optical splitter (5) afterwards, beam of laser is passed through apart from L direct irradiations in the sample (6) surface, another
Shu Jiguang reaches the level crossing (8) after apart from a, and the sample (6) table is irradiated in after the level crossing (8) reflection
Face, the above-mentioned two beam laser for being irradiated in the sample (6) surface are angle o, and the dry of light can be produced at the sample (6) surface
Pattern is related to, it is described adjustable apart from a and L, so as to change sample (6) the Surface Interference bar by the value for changing the angle θ
Line spacing i；The level crossing (8) is connected with the displacement platform (7) and position modulation can be carried out to the level crossing (8),
So that the level crossing (8) carries out sinusoidal vibration along its normal orientation with frequency 800Hz, so as to change the sample (6) surface
Locate position of interference fringe.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107024415A (en) * | 2017-04-17 | 2017-08-08 | 金华职业技术学院 | A kind of device for studying molecular migration motion |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107024415A (en) * | 2017-04-17 | 2017-08-08 | 金华职业技术学院 | A kind of device for studying molecular migration motion |
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