CN109324406A - Plant living cell capture and manipulation device and method based on fiber optic probe - Google Patents
Plant living cell capture and manipulation device and method based on fiber optic probe Download PDFInfo
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- CN109324406A CN109324406A CN201811354421.9A CN201811354421A CN109324406A CN 109324406 A CN109324406 A CN 109324406A CN 201811354421 A CN201811354421 A CN 201811354421A CN 109324406 A CN109324406 A CN 109324406A
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- 239000000523 sample Substances 0.000 title claims abstract description 121
- 238000000034 method Methods 0.000 title claims abstract description 35
- 239000000835 fiber Substances 0.000 title claims description 14
- 239000013307 optical fiber Substances 0.000 claims abstract description 141
- 210000004027 cell Anatomy 0.000 claims abstract description 72
- 239000011521 glass Substances 0.000 claims abstract description 32
- 230000003287 optical effect Effects 0.000 claims description 44
- 238000002474 experimental method Methods 0.000 claims description 19
- 238000003491 array Methods 0.000 claims description 15
- 210000003763 chloroplast Anatomy 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 229920000742 Cotton Polymers 0.000 claims description 4
- 238000005253 cladding Methods 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims 1
- 239000011247 coating layer Substances 0.000 claims 1
- 230000004927 fusion Effects 0.000 claims 1
- 210000003463 organelle Anatomy 0.000 abstract description 17
- 238000010353 genetic engineering Methods 0.000 abstract description 3
- 230000029553 photosynthesis Effects 0.000 abstract description 2
- 238000010672 photosynthesis Methods 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 23
- 230000000694 effects Effects 0.000 description 13
- 238000012576 optical tweezer Methods 0.000 description 8
- 238000000399 optical microscopy Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 238000002844 melting Methods 0.000 description 6
- 230000008018 melting Effects 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- 230000004899 motility Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 244000052355 Hydrilla verticillata Species 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 230000003685 thermal hair damage Effects 0.000 description 1
Classifications
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/32—Micromanipulators structurally combined with microscopes
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- G—PHYSICS
- G21—NUCLEAR PHYSICS; NUCLEAR ENGINEERING
- G21K—TECHNIQUES FOR HANDLING PARTICLES OR IONISING RADIATION NOT OTHERWISE PROVIDED FOR; IRRADIATION DEVICES; GAMMA RAY OR X-RAY MICROSCOPES
- G21K1/00—Arrangements for handling particles or ionising radiation, e.g. focusing or moderating
- G21K1/006—Manipulation of neutral particles by using radiation pressure, e.g. optical levitation
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The plant living body cell capture and control device and its method that the invention discloses a kind of based on optical fiber probe.Plant living body cell capture and control device based on optical fiber probe include laser, computer, charge coupled cell, microscope, object lens, optical fiber probe, glass tube, glass slide, optical fiber adjusting bracket, objective table.The method of plant living body cell capture based on optical fiber probe and manipulation, the specific steps are as follows: step S1: optical fiber probe of the preparation for capturing and manipulating；Step S2: one-dimensional chloroplaset array is assembled；Step S3: it manipulates one-dimensional chloroplaset array and is moved in living cells；Step S4: two-dimentional chloroplaset array is assembled.The present invention realizes manipulation, arrangement and the assembling to the typical organelle chloroplaset in plant cell, this is most important to the photosynthesis and genetic engineering of plant.
Description
Technical field
The invention belongs to optical technical field, more particularly to a kind of plant living body cell capture based on optical fiber probe and
Control device and its method.
Background technique
Traditional optical tweezer can cell to living body surface layer contactless capture and manipulation, but its penetration depth is limited, and
And there is complicated, expensive device and huge optical system, can not go deep into organism to the cell of deep layer carry out capture and
Manipulation.
Existing internal control method may be implemented to go deep into the capture of organism inner cell and manipulation, utilize laser pair
Perhaps particle generates the focal point that cell or particle are pulled to field gradient power by gradient force to cell, completes capture and manipulation, still
Individual cells can only be captured or space is fixed, it is difficult to while multiple cells even organelle of cell interior are carried out
Manipulation, arrangement and assembling.This is because traditional light manipulates technology, such as traditional optical tweezer, it is the control device based on single beam,
An object can only be once manipulated, also, since traditional optical tweezer relies on the micro- lens head of high-NA (NA:1.0-1.4),
It is difficult to protrude into inside living body, therefore, it is difficult to manipulate the cell of organism deep layer and be difficult to even intracellular to multiple cells simultaneously
The organelle in portion is manipulated, arranged and is assembled.
Summary of the invention
The purpose of the present invention is to provide a kind of plant living body cell capture and control device based on optical fiber probe, solution
The limited penetration depth of traditional optical tweezer and the problem of huge optical system in the prior art.
Another object of the present invention is to provide the sides of a kind of the plant living body cell capture based on optical fiber probe and manipulation
Method, the intracorporal cell capture of biology and manipulation can not be goed deep into and be difficult to simultaneously to multiple by solving traditional optical tweezer in the prior art
The problem of organelle of cell even cell interior is manipulated, arranged and is assembled.
The technical scheme adopted by the invention is that plant living body cell capture and control device based on optical fiber probe, packet
Optical microscopy is included, optical microscopy is made of charge coupled cell, microscope, object lens, and microscopical top is provided with charge
Coupling element, charge coupled cell are connect with pcs signal, and object lens are arranged right below objective table, and objective table, which is equipped with, carries glass
One end of piece, optical fiber probe is placed on glass slide, and the other end connecting laser of optical fiber probe, optical fiber probe is close to objective table
Side outside be cased with glass tube, glass tube is fixed on optical fiber adjusting bracket.
Further, the II-DH II of optical microscopy model HISOMET；
The laser is that wavelength is 980 nanometers of single-mode lasers；
The amplification factor of the object lens is 40 ~ 100 times, and numerical aperture is 0.25 ~ 0.73, and operating distance is 1.0 ~ 3.0 millimeters.
Further, the mobile accuracy of the optical fiber adjusting bracket is 50~60 nanometers, and the mobile accuracy of the objective table is
50~60 nanometers.
Further, the diameter of the fiber optic probe tip be 700 nanometers, length be 6.1 microns, cone angle be 74 ° ±
5°；
The internal diameter of the glass tube is 0.9 millimeter, and 0.1 millimeter of wall thickness, length is 12 centimetres.
Another technical solution adopted in the present invention is that plant living body cell capture and manipulation based on optical fiber probe
Method, the specific steps are as follows:
Step S1: optical fiber probe of the preparation for capturing and manipulating: optical fiber probe is drawn by the single mode optical fiber of a standard through melting
Cone method is prepared；
Step S2: one-dimensional chloroplaset array is assembled；
Step S3: one-dimensional chloroplaset array is manipulated in plant living body intracellular motility；
Step S4: two-dimentional chloroplaset array is assembled.
Further, the drawing cone method specifically follows the steps below:
Step S11, after peelling off one section 2 centimetres of coat among single mode optical fiber, single mode optical fiber is packed into a glass tube；
Step S12, exposed single mode optical fiber is placed in parallel at the flame envelope above alcolhol burner, stands 5-10 seconds to single mode optical fiber
After melting, the part of melting is drawn into as 700 nanometers by diameter with 3-5mm/s speed, length is 6.1 microns, and cone angle is 74 °
± 5 ° of optical fiber probe；
Single mode optical fiber core diameter is 9 microns, and cladding diameter is 125 microns, and connector type is FC/PC.
Further, the step S2 is specifically followed the steps below:
Step S21, optical fiber probe is fixed on optical fiber adjusting bracket, after then wrapping up the root of determinand leaf with wet cotton
It is placed on glass slide, glass slide is placed horizontally on objective table；
Step S22, leaf is paved on glass slide with tweezers, then water is dripped on leaf；
Step S23, fiber optic probe tip is placed on above leaf, optical fiber probe does not contact leaf, then into optical fiber probe
Being passed through power is 30-50 milliwatt, and the laser that wavelength is 980 nanometers manipulates 3 ~ 6 chloroplasets and is assembled into one-dimensional array in an orderly manner.
Further, the step S3 is specifically followed the steps below:
Step S31, the one-dimensional array in optical fiber probe operating steps S23 is placed on the middle part of cell, then uses optical fiber adjusting bracket
By optical fiber probe with the speed of average 3.4 ± 0.2 micron per minutes to-yThe direction of axis is mobile.
Further, the step S4 is specifically followed the steps below:
Step S41, with the speed of average 3.4 ± 0.2 micron per minutes to+yAxis direction moving fiber probe, one-dimensional chloroplaset battle array
Column with optical fiber probe movement also to+yAxis direction is mobile, when one-dimensional chloroplaset array is moved near free chloroplaset
When, free chloroplaset forms the second one-dimensional chloroplaset array of string, and it is mobile that the second one-dimensional chloroplaset array of string tends to optical axis center
In the process, the second one-dimensional chloroplaset array of string is steadily arranged in the upside of optical axis center, forms stable two-dimentional battle array in an orderly manner
Column；
Step S42, laser continues to that experiment terminates, chloroplaset mixed and disorderly distribution in the cell again after laser is closed.
Further, water covering leaf is 1 ~ 2 centimetre in the step S22；
Optical fiber probe is placed in above leaf 3 microns in the step S23.
The beneficial effects of the present invention are: 1. the present invention is based on the plant living body cell capture and control device of optical fiber probe are non-
Often flexibly and miniaturization, the problem of avoiding the limited penetration depth of traditional optical tweezer and huge optical system, therefore can be deep
Enter living body trapped inside and manipulation cell.
2. mini optical fibre probe of the present invention is in the case where interfering living body normal activities, while more to living body inside
The organelle of a cell even cell interior is manipulated, arranged and is assembled.Close contact between organelle can promote thin
Cell function is coordinated in communication between born of the same parents' device, and ion (such as Ca ion) between organelle and lipid provide channel.This
Invention realizes manipulation, arrangement and assembling to the typical organelle chloroplaset in plant cell, this photosynthetic work to plant
With most important with genetic engineering.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
It obtains other drawings based on these drawings.
Fig. 1 is plant living body cell capture and control device figure based on optical fiber probe.
Fig. 2 be plant living body cell capture and control method based on optical fiber probe embodiment in assemble one-dimensional chloroplaset
Array experiment result figure.
Fig. 2 a is the experiment photo figure that optical fiber probe captures that 3 chloroplasets are assembled into one-dimensional array.
Fig. 2 b is the experiment photo figure that optical fiber probe captures that 4 chloroplasets are assembled into one-dimensional array.
Fig. 2 c is the experiment photo figure that optical fiber probe captures that 5 chloroplasets are assembled into one-dimensional array.
Fig. 2 d is the experiment photo figure that optical fiber probe captures that 6 chloroplasets are assembled into one-dimensional array.
Fig. 3 be plant living body cell capture and control method based on optical fiber probe embodiment in transport one-dimensional chloroplaset
Array experiment result figure.
Fig. 3 a istOptical fiber probe captures the experiment photo figure that 4 chloroplasets are assembled into one-dimensional array when=0 s.
Fig. 3 b ist When=3 s optical fiber probe manipulate one-dimensional chloroplaset array to-yThe mobile experiment photo figure of axis direction.
Fig. 3 c ist When=5.8 s optical fiber probe manipulate one-dimensional chloroplaset array to+yThe mobile experiment photo of axis direction
Figure.
Fig. 4 is that the present invention is based on two dimension is assembled in the embodiment of the plant living body cell capture of optical fiber probe and control method
Chloroplaset array of figure.
Fig. 4 a1 is the schematic diagram of 3 × 4 two-dimentional chloroplaset array assembling process.
Fig. 4 a2 is the experiment displaing micro picture of 3 × 4 two-dimentional chloroplaset array assembling process.
Fig. 4 b1 is the schematic diagram that 3 × 4 two-dimentional chloroplaset arrays assemble result.
Fig. 4 b2 is the experiment displaing micro picture that 3 × 4 two-dimentional chloroplaset arrays assemble result.
Fig. 4 c1 is the experiment displaing micro picture for assembling 2 × 2 chloroplaset arrays.
Fig. 4 c2 is the experiment displaing micro picture for assembling 5 × 2 chloroplaset arrays.
In figure, 1. lasers, 2. computers, 3. charge coupled cells, 4. microscopes, 5. object lens, 6. optical fiber probes, 7. glass
Pipe, 8. glass slides, 9. optical fiber adjusting brackets, 10. objective tables.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
Plant living body cell capture and control device based on optical fiber probe, as shown in Figure 1, including optical microscopy, light
It learns microscope to be made of charge coupled cell 3, microscope 4, object lens 5, the top of microscope 4 is provided with charge coupled cell 3, electricity
Lotus coupling element 3 is connect with 2 signal of computer, and object lens 5 are arranged right below objective table 10, and objective table 10 is equipped with glass slide 8, light
One end of fine probe 6 is placed on glass slide 8, and the other end connecting laser 1 of optical fiber probe 6, optical fiber probe 6 is close to objective table
Glass tube 7 is cased with outside 10 side, glass tube 7 is fixed on optical fiber adjusting bracket 9.
Optical microscopy is the optical microscopy of a transmission-type, II-DH II of model HISOMET.
Laser 1 is that wavelength is 980 nanometers of single-mode lasers, and for capture and manipulation cell, the laser of the wavelength is to life
The absorption of object tissue is low, can be effectively prevented from the thermal damage as caused by light absorption, and this wavelength is living suitable for plant
The manipulation of body light.
Charge coupled cell 3, for obtaining the image and video of experiment from computer 2.
The amplification factor of object lens 5 is 40 ~ 100 times, and numerical aperture, that is, NA is 0.25 ~ 0.73, and operating distance is 1.0 ~ 3.0 millis
Rice, for observing and recording experimentation.
The mobile accuracy of optical fiber adjusting bracket 9 is 50~60 nanometers, and objective table 10 can be three-dimensional mobile, mobile accuracy is 50~
60 nanometers.
The internal diameter of glass tube 7 is 0.9 millimeter, and 0.1 millimeter of wall thickness, length is 12 centimetres.
Optical fiber probe 6 is prepared by fused biconical taper method, and the diameter at tip is 700 nanometers, and length is 6.1 microns, cone angle
It is 74 ° ± 5 °.Cone angle has good convergence to act in 6 pairs of light of optical fiber probe of this range, penetrates readily through biological tissue, is suitble to
For the manipulation of living body, and the optical fiber of the range cone angle is conducive to the cell or organelle that capture diameter is 1 ~ 10 micron, when
When cone angle is greater than 79 °, the ability of the tip converged light of optical fiber probe 6 weakens significantly, is unfavorable for capture and manipulation cell or thin
Born of the same parents' device, when cone angle is less than 69 °, it is difficult to capture 1 ~ 10 micron of diameter of organelle or cell.
In experiment, leaf is wrapped into root with wet cotton to keep activity.
It opens in the laser importing fiber probe 6 that a branch of wavelength is 980 nanometers by laser 1, light will be coupled into optical fiber spy
In needle 6, optical fiber probe 6 focuses on light on optical axis and transmits along optical axis, and the organelle chloroplaset of cell is due to light in leaf
The effect of gradient force is trapped in one by one on the optical axis direction of optical fiber probe 6, to realize that capture and manipulation are multiple
Object, and form one-dimensional chloroplaset array.
The present invention is based on the plant living body cell capture and control device of optical fiber probe are simple, just with single big cone angle
Optical fiber probe 6 and foil, it will be able to which realization carries out the organelle in the cell of living body inside with optical method non-
Contact, undamaged manipulation, assembling, transport.Bulky compared to traditional optical tweezer, the device volume is small, simple and flexible and easily
In integrated, and the small problem of traditional optical tweezer penetration depth is overcome, while its compact structure can meet living body well
The requirement of manipulation.
In order to solve the problems, such as that the organelle to multiple cells even cell interior is manipulated, arranged and assembled, herein
Light is focused on optical axis by optical fiber probe 6 and is transmitted along optical axis, in this way to improve the number of manipulation object, is manipulated simultaneously
Multiple objects, also, since optical fiber probe 6 has the advantages that miniaturization, integrated and flexibility, it can be implanted into living body, to life
The cell or organelle of object living body deep inside are manipulated.
The method of plant living body cell capture based on optical fiber probe and manipulation, the specific steps are as follows:
Step S1: optical fiber probe 6 of the preparation for capturing and manipulating: optical fiber probe 6 is by the single mode optical fiber of a standard through melting
It draws cone method to be prepared, specifically follows the steps below:
Step S11, after peelling off one section 2 centimetres of coat among single mode optical fiber, single mode optical fiber is packed into a glass tube 7
For protecting single mode optical fiber, the internal diameter of glass tube 7 is 0.9 millimeter, and 0.1 millimeter of wall thickness, length is 12 centimetres；
Step S12, exposed single mode optical fiber is placed in parallel at the flame envelope above alcolhol burner, stands 5-10 seconds to single mode optical fiber
After melting, the part of melting is drawn into as 700 nanometers by diameter with 3-5mm/s speed, length is 6.1 microns, and cone angle is 74 °
± 5 ° of optical fiber probe 6；
Single mode optical fiber core diameter is 9 microns, and cladding diameter is 125 microns, and connector type is FC/PC；
The diameter and cone angle of optical fiber probe 6 are regulated and controled by the speed that control is drawn；
Step S2: one-dimensional chloroplaset array is assembled；
Step S21, optical fiber probe 6 is fixed on optical fiber adjusting bracket 9, then wraps up the root of determinand leaf with wet cotton
After be placed on glass slide 8, glass slide 8 is placed horizontally on objective table 10；
Step S22, leaf is paved on glass slide 8 with tweezers, then water is dripped on leaf；Dripping water keeps leaf
Activity；
Water covering leaf is 1 ~ 2 centimetre in step S22, is lower than 1 centimetre, it will the activity for influencing leaf is unfavorable for higher than 2 centimetres
The observation of optical microscopy；
Step S23,6 tip of optical fiber probe is placed on above leaf, optical fiber probe 6 does not contact leaf, then toward optical fiber probe 6
In to be passed through power be 30-50 milliwatt, wavelength is 980 nanometers of laser, and experimental result show that finally to manipulate 3 ~ 6 chloroplasets orderly
Ground is assembled into one-dimensional array；
Optical fiber probe 6 is placed in above leaf 3 microns in step S23, is conducive to manipulate the organelle inside leaf, while guaranteeing to swash
The fuel factor that light generates does not damage biological cell；
The power that optical fiber probe 6 is passed through in step S23 defies capture organelle if too low, and power is excessive to generate thermal effect
It answers；Due toyThe effect of optical gradient forces in axis direction, the chloroplaset to dissociate around optical fiber probe 6 being caught one by one
Obtain the optical axis direction in optical fiber probe 6, the chloroplaset on optical axis due to byxThe optical gradient forces of axis direction will tightly be bound
Together, the number for manipulating chloroplaset is related with the watt level for being passed through optical fiber probe 6；
Step S3: one-dimensional chloroplaset array is manipulated in plant living body intracellular motility；
Step S31, the one-dimensional array in 6 operating steps S23 of optical fiber probe is placed on the middle part of cell, then uses optical fiber adjusting bracket
9 by optical fiber probe 6 with the speed of average 3.4 ± 0.2 micron per minutes to-yThe direction of axis is mobile；
In step S31 by optical fiber probe 6 with the speed of average 3.4 ± 0.1 micron per minutes to+yThe direction of axis is mobile, equally,
To+yWhen the direction of axis is mobile, 6 maximum speed of moving fiber probe is 7.5 ± 0.2 micron per minutes, maximum fast when being greater than this
When spending, chloroplaset will be detached from optical axis without the control by optical gradient forces, and when being less than the maximum speed, chloroplaset is by light gradient
Power stablizes capture, and optical fiber probe 6 is followed to move together；
Since light captures stability, it is not detached from from the optical axis of optical fiber probe 6 in moving process Chloroplast, due to chloroplaset quilt
It is strapped on the optical axis of optical fiber probe 6, therefore chloroplaset one-dimensional array portion's fortune in the cell is controlled by moving fiber probe 6
It is dynamic；
Step S4: two-dimentional chloroplaset array is assembled；
Step S41, with the speed of average 3.4 ± 0.2 micron per minutes to+yAxis direction moving fiber probe 6, one-dimensional chloroplaset
Array with optical fiber probe 6 movement also to+yAxis direction is mobile, when to be moved to free chloroplaset attached for one-dimensional chloroplaset array
When close, free chloroplaset due to by+yThe effect of axis direction gradient force can tend to optical axis center and move down, and due to
ByxThe effect of axis direction gradient force, free chloroplaset to form the one-dimensional chloroplaset battle array of the second string by stable binding together
Column, the second one-dimensional chloroplaset array of string tend in optical axis center moving process, and the second one-dimensional chloroplaset array of string can go here and there to first
One-dimensional chloroplaset array generates downward thrust, and the first one-dimensional chloroplaset array of string will deviate slightly downwards from optical axis direction,
Interaction force between the two one-dimensional chloroplaset arrays of string is arranged closely in optical axis center two sides, the first string one after reaching balance
Dimension chloroplaset array is steadily arranged in the downside of optical axis center, and the second one-dimensional chloroplaset array of string is steadily arranged in optical axis
The upside of the heart forms stable two-dimensional array with this in an orderly manner；
By changing the power of 980 nm laser of wavelength, it is one-dimensional that capture 3 ~ 6 chloroplasets of manipulation are assembled into the first string in an orderly manner
Chloroplaset array is then green by the free one-dimensional leaf of the second string of chloroplaset formation of the effect capture different number of optical gradient forces again
Volume array, two string one-dimensional arrays are formed and are closely arranged near optical axis center, complete the assembling of two-dimentional chloroplaset array；
Step S42, laser 1 continues to that experiment terminates, chloroplaset mixed and disorderly distribution in the cell again after laser 1 is closed.
Embodiment 1
The live plant that the present embodiment uses is aquatic algae, and scientific name is hydrilla verticillata, and the leaf of the plant is transparent,
Convenient for studying the characteristic of chloroplaset.
After laser importing fiber probe 6, due toyThe effect of optical gradient forces in axis direction dissociates around optical fiber probe 6
Chloroplaset will be captured and be limited on the optical axis of optical fiber probe 6, the chloroplaset on 6 optical axis of optical fiber probe due to byxAxis direction gradient force, to form one-dimensional chloroplaset array in optical axis direction.
As shown in Figure 2 a, after being passed through power into optical fiber probe 6 as 30 milliwatts, 980 nanometers of wavelength of laser, 3 leaves are green
Body is captured to be assembled into one-dimensional array.
As shown in Figure 2 b, after being passed through power into optical fiber probe 6 as 35 milliwatts, 980 nanometers of wavelength of laser, 4 leaves are green
Body is captured to be assembled into one-dimensional array.
As shown in Figure 2 c, after being passed through power into optical fiber probe 6 as 42 milliwatts, 980 nanometers of wavelength of laser, 5 leaves are green
Body is captured to be assembled into one-dimensional array.
As shown in Figure 2 d, after being passed through power into optical fiber probe 6 as 50 milliwatts, 980 nanometers of wavelength of laser, 6 leaves are green
Body is captured to be assembled into one-dimensional array.
Being passed through power in this experiment into optical fiber probe 6 is 35 milliwatts, the laser that wavelength is 980 nanometers, final manipulation 4
Chloroplaset is assembled into one-dimensional chloroplaset array in an orderly manner, and the chloroplaset long axis of cell interior is 2.3 microns, and short axle is 1.2 micro-
Rice；Chloroplaset is captured with optical fiber probe 6 and is assembled into one-dimensional array, as shown in Figure 3a, is then visited optical fiber with optical fiber adjusting bracket 9
Needle 6 with the speed of average 3.4 micron per minutes to-yThe direction of axis is mobile, one-dimensional chloroplaset array within the time of 3s to-yAxis side
To the distance for being shifted 10.3 microns, as shown in Figure 3b, then by optical fiber probe 6 to+yAxis direction is mobile, one-dimensional chloroplaset battle array
Be listed in time of 2.8s to+yAxis direction is shifted 9.5 microns of distance, as shown in Figure 3c, during entire mobile,
There is no chloroplaset to be detached from from optical axis, it was demonstrated that the stability of light capture.
Two one-dimensional chloroplaset arrays are merged by optical fiber probe 6, realize orderly two-dimentional chloroplaset
The assembling of array.After being passed through the laser that power is 35 milliwatts, wavelength is 980 nanometers in optical fiber probe 6,4 free leaves
Green body is steadily assembled into one-dimensional array, then with the speed of average 3.4 ± 0.2 micron per minutes to+yThe mobile light in the direction of axis
Fine probe 6, free chloroplaset due to by+yThe effect of axis direction gradient force can tend to optical axis center and move down, and by
In byxThe effect of axis direction gradient force, it is green that 3 free chloroplasets by stable binding together to form the one-dimensional leaf of the second string
Volume array, the second one-dimensional chloroplaset array of string tend in optical axis center moving process, and the second one-dimensional chloroplaset array of string can be to the
A string of one-dimensional chloroplaset arrays generate downward thrust, and the first one-dimensional chloroplaset array of string will be inclined slightly downwards from optical axis direction
It moves, the interaction force between two string one-dimensional arrays is arranged closely in optical axis center two sides after reaching balance, and the first string is one-dimensional
Chloroplaset array is steadily arranged in the downside of optical axis center, and the second one-dimensional chloroplaset array of string is steadily arranged in optical axis center
Upside, form stable two-dimensional array in an orderly manner with this, the two dimension chloroplaset array assembling process as shown in Fig. 4 a1- Fig. 4 a2,
Manipulation is moved near 3 free chloroplasets by the one-dimensional array that 4 chloroplasets assemble, finally by two one-dimensional arrays
Movement merges, and realizes the assembling of orderly two-dimentional chloroplaset array, as shown in Fig. 4 b1- Fig. 4 b2,6 success of optical fiber probe
Assemble 3 × 4 arrays.By the chloroplaset of capture different number, the array of different size is assembled, if shown in Fig. 4 c1- figure c2,
2 × 2 and 5 × 2 arrays are successfully assembled respectively.
The present invention realizes manipulation, arrangement and the assembling of the typical organelle chloroplaset intracellular to plant living body, this
It is most important to the photosynthesis and genetic engineering of plant.
Each embodiment in this specification is all made of relevant mode and describes, same and similar portion between each embodiment
Dividing may refer to each other, and each embodiment focuses on the differences from other embodiments.Especially for system reality
For applying example, since it is substantially similar to the method embodiment, so being described relatively simple, related place is referring to embodiment of the method
Part explanation.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the scope of the present invention.It is all
Any modification, equivalent replacement, improvement and so within the spirit and principles in the present invention, are all contained in protection scope of the present invention
It is interior.
Claims (10)
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| CN113238075A (en) * | 2021-04-22 | 2021-08-10 | 哈尔滨工程大学 | Flow velocity meter based on optical fiber tweezers technology |
| CN114112909A (en) * | 2021-11-16 | 2022-03-01 | 暨南大学 | Application of chloroplasts as microlenses, microscopy imaging, fluorescence enhancement |
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