CN113481095A

CN113481095A - Precise active optical control method and device based on double-core optical fiber living body single cell rotation

Info

Publication number: CN113481095A
Application number: CN202110782406.XA
Authority: CN
Inventors: 尹君; 陈宏宇; 于凌尧; 王少飞; 贾源; 胡徐锦; 苑立波
Original assignee: Guilin University of Electronic Technology
Current assignee: Guilin University of Electronic Technology
Priority date: 2021-07-12
Filing date: 2021-07-12
Publication date: 2021-10-08

Abstract

The present invention provides a method and device for precise active light manipulation of single-cell rotation in a living body based on a double-core optical fiber. It is characterized in that the device is composed of two double-core optical fiber optical manipulation systems and a microscopic imaging system. The two output ends are processed into a double-core optical fiber with a specific angle truncated cone. The laser beam output from one of the dual-core fibers forms a Bessel light field near the output end to achieve stable capture of cells. The other two-core fiber outputs laser pulses with a certain time interval, which act on both ends of the captured cells respectively. Adjust the intensity and time interval of the two pulses to achieve precise active light manipulation of the rotation angle of the captured cells. The invention can realize precise active light manipulation of stable capture and rotation angle of specific living single cells, has the characteristics of low cost, high flexibility and high integration, and has wide application prospects in research fields such as medicine, biology and life sciences. .

Description

Precise active optical control method and device based on double-core optical fiber living body single cell rotation

(I) technical field

The living body single cell rotation accurate active optical control method and the living body single cell rotation accurate active optical control device based on the double-core optical fiber can be used for realizing stable capture of specific living body single cells and accurate active optical control of the living body single cells around a specific rotation axis. Belonging to the field of photodynamic manipulation research.

(II) background of the invention

Since the new century, with the continuous progress of scientific means, the exploration of the nature by human beings is also deepened, and the understanding of the organism by people is also deepened gradually, and the research of people is deepened from the research of the nature to individuals, from individuals to cells, from the research of the cells to molecules. Among them, the cell is used as a basic unit of the vital structure and function, and intensive research on the cell is a key to reveal the phenomenon of life, conquer diseases and modify life. "all-life phenomenon's secret from the cell is sought after" as pointed out by EBWilson, a well-known biological specialist. The research on the cells is continuously deepened and developed along with the scientific development law, and the cells are subjected to different research levels such as a cell level, a subcellular level and a molecular level.

With the progress of research, reliable scientific bases are provided for revealing the essence and basic rules of life activities at a deeper level, and the challenge is to take large environmental living cells for developing life activities as test tubes and obtain subcellular fine structures of living unicells in a non-contact and non-destructive manner on the premise of avoiding influencing the properties of the cells and the microenvironment where the cells are located as far as possible. This requires stable capture and precise manipulation of living single cells during long-term observation and study.

In 1970, AAshkin et al first proposed the use of a focused laser beam to capture fine particles by theoretical analysis and simulation calculations. In 1986, AAshkin et al realized an optical tweezers technique for stably trapping minute particles with a stable energy trap formed by a focused laser beam. The optical tweezers technology utilizes an optical trap formed by a focusing optical field to generate a mechanical effect, and can stably capture, accurately control and rapidly screen single viruses, cells and even biomacromolecules in a non-mechanical contact and non-damage mode under the condition of not influencing the interior of cells and the microenvironment where the cells are located. In addition, the optical tweezers can not only control the particles, but also measure the tiny force. As a quantitative analysis tool, the optical tweezers technology can apply a calibrated piconiu-level optical trapping force to a system of interest, so that the displacement of a target system caused by the action of the optical trapping force can be measured with high precision and high sensitivity. At present, in the research fields of molecular biology, biochemistry, biophysics and the like, the optical tweezers analysis technology has been developed into a powerful tool for controlling and analyzing cells at the molecular level, and is widely applied to numerous research fields of biomechanics, biopolymers, biomacromolecules, molecular motors and the like. Meanwhile, the proposal and development of the optical tweezers technology opens the door for observing living cells in a liquid environment in a non-mechanical contact and non-destructive manner for a long time to obtain the internal structure and the physical and chemical properties of the living cells, and further deeply researching the biological regulation and control mechanism and the like of the cell life activity process.

The traditional optical tweezers technology not only needs to use a large-numerical-aperture microscope objective and a complex optical path system, but also is limited by a plurality of factors such as working distance, substrate compatibility and the like in the use process, and the system has large volume, high manufacturing cost and poor flexibility. In recent years, the fiber optical tweezers technology based on fiber implementation can perform accurate control such as capture, stretching, moving, rotating and the like on living single cells in a liquid environment. The optical fiber has small volume and light weight, can flexibly move in a medium at will, and the optical fiber tweezers system realized by the optical fiber has simple structure, strong operability, high integratability and flexibility.

The invention discloses a living body single cell rotation accurate active optical control method and a living body single cell rotation accurate active optical control device based on a double-core optical fiber. On the premise that the environment of the cell does not influence the property of the cell, the living unicells in the liquid environment are stably captured in a non-contact and non-destructive mode, and the rotation angle of the living unicells is accurately and actively controlled by light. The method and the system have the characteristics of simple structure, low manufacturing cost, high flexibility, high integration level and the like, and have wide application prospects in the research fields of medicine, biology, life science and the like.

Disclosure of the invention

The invention aims to provide a living body single cell stable capturing and rotation angle accurate active optical control method and device based on a double-core optical fiber, and the method and device have the characteristics of simple structure, convenience in operation, high flexibility, high integration level and the like.

The purpose of the invention is realized as follows:

the device is mainly formed by oppositely installing two double-core

optical fibers

10 and 19, wherein the central axes of two fiber cores are vertical to each other, and the output end faces of the two double-core optical fibers are processed into specific angles. The device comprises a continuous working laser 1, an output coupling optical fiber 2, two-port optical

fiber beam splitters

3, 6 and 12, single-mode

optical fibers

4, 5, 7, 8, 13 and 14, optical fiber beam combiners 9 and 18, an optical fiber frequency modulator 11, optical

fiber intensity modulators

15 and 16, an optical fiber time delayer 17, double-core

optical fibers

10 and 19 of which the output ends are processed into frustum with a specific angle, a white light LED light source 20, a 45-degree reflector 21, a condenser 22, a apochromatic microscope objective 23, an optical filter 24, a CMOS camera 25 and a cell to be detected 26. Laser beams emitted by a continuous working laser 1 in the system are coupled into a two-port optical fiber beam splitter 3 through an output coupling optical fiber 2 and are respectively coupled into single-mode optical fibers 4 and 5 according to a certain intensity ratio. The laser beam transmitted by the single mode fiber 5 is divided into two beams of laser with equal intensity by the two-port fiber beam splitter 6, and the two beams of laser are transmitted by the single mode fibers 7 and 8, coupled by the fiber beam combiner 9, and processed into the double-core fiber 10 with the output end being processed into the frustum with a specific angle, so that the stable capture of the cell to be detected is realized. The laser beam passing through the single mode fiber 4 is frequency modulated by the fiber frequency modulator 11 to generate a laser pulse with adjustable pulse width, and then is divided into two beams of laser with equal intensity by the two-port fiber beam splitter 12, and the two beams of laser are respectively coupled into the

single mode fibers

13 and 14. The laser pulses coupled into the single mode fiber 13 are pulse intensity modulated via a fiber intensity modulator 16. The laser pulses coupled into the single mode fiber 14 are pulse intensity modulated by a fiber intensity modulator 15 and a certain time delay is introduced by a fiber time delay 17. Two laser pulses with certain time delay are coupled into and output from the optical fiber beam combiner 18 to be processed into the double-core optical fiber 19 of the frustum with a specific angle, so that accurate active control of the cell to be detected is realized. The white light emitted by the white light LED light source 20 is reflected by the 45-degree reflector 21, and then forms a full-field uniform kohler illumination on the sample 26 to be measured by the condenser 22. The image information of the sample 26 to be measured is collected by the apochromatic microscope objective 23, the background noise is eliminated by the optical filter 24, and the spatial position information of the cell is recorded by the CMOS camera 25.

The main part of the light control device is composed of two double-core

optical fibers

10 and 19. The optical fiber 10 is used for fixing cells, and the optical fiber 19 is used for manipulating the cells and enabling the cells to rotate. Fig. 2(a) is a schematic diagram of the position and arrangement of two dual-core optical fibers: the optical fiber for fixing the cells is transversely arranged, and the optical fiber for controlling the cells is longitudinally arranged.

As shown in fig. 2 (a): two double-core optical fibers are oppositely arranged, laser emitted by the continuous working laser 1 is divided into two beams with different intensity proportions by the two-port beam splitter 3, wherein the intensity of a light beam entering the single-mode optical fiber 5 is smaller than that of a light beam entering the single-mode optical fiber 4, and the light intensity for capturing cells is larger than that of light for controlling the cells, so that the cells are prevented from being separated from a control range due to insufficient captured light intensity when the cells are controlled. The light beam in the single mode fiber 5 is split into two beams by a beam splitter 6, and is input into a double-core fiber 10 by a fiber combiner 9. The light is used for stably capturing the cells and fixing the cells. Light in the single-mode fiber 4 is modulated by the frequency modulator 11 to become pulse light, the pulse light is divided into two beams by the beam splitter and finally output by the double-core fiber 19, and the light is used for controlling cells.

The optical fiber 13 and the optical fiber 14 each have an

intensity modulator

15, 16, wherein the light in the optical fiber 14 is used to push the cell to rotate, the light in the optical fiber 13 is used to stop the cell from rotating, wherein the light modulated by the intensity modulator 15 is slightly smaller than the intensity modulator 16, and the light modulated by the intensity modulator 15 is generated for a time interval by the time delay device 17.

(IV) description of the drawings

FIG. 1 is a schematic structural diagram of a cell rotation optical manipulation method and device based on a dual-core optical fiber.

FIG. 2(a) is a schematic diagram showing the main structure of light manipulation and the manner of light manipulation, showing a section view and the internal structure of a two-core optical fiber, which is formed by two twin-wire optical fibers disposed opposite to each other, for fixing cells and manipulating cells, respectively. FIG. 2(b) shows a cut-away view of a two-core optical fiber.

Fig. 3 is a power diagram of pulsed light for cell manipulation and fixation.

Description of reference numerals: 1-a continuous laser; 2-single mode fiber; 3-a fiber optic splitter; 4-single mode fiber; 5-single mode fiber; 6-fiber beam splitter; 7-a single mode optical fiber; 8-single mode fiber; 9-an optical fiber combiner; 10-a dual core optical fiber; 11-a fiber frequency modulator; 12-a fiber optic splitter; 13-a single mode optical fiber; 14-a single mode optical fiber; 15-fiber intensity modulator; 16-a fiber intensity modulator; 17-fiber time delay; 18-an optical fiber combiner; 19-a dual core fiber; 20-a white light source; a 21-45 degree mirror; 22-a condenser lens; 23-apochromatic microobjective; 24-an optical filter; 25-CMOS camera; 26-test cells.

(V) detailed description of the preferred embodiments

The present invention is further described in detail below with reference to examples to enable those skilled in the art to practice the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

A cell rotation accurate optical control method and device based on a double-core optical fiber. The method is characterized in that: the device is mainly formed by oppositely installing double-core

optical fibers

10 and 19, wherein the central lines of two fiber cores are vertical to each other, and the output end face of the double-core optical fibers is processed into a specific angle. The device comprises a continuous working laser 1, an output coupling optical fiber 2, two-port optical

fiber beam splitters

3, 6 and 12, single-mode

optical fibers

fiber intensity modulators

15 and 16, an optical fiber time delayer 17, double-core

optical fibers

10 and 19 of which the output ends are processed into frustum with a specific angle, a white light LED light source 20, a 45-degree reflector 21, a condenser 22, a apochromatic microscope objective 23, an optical filter 24, a CMOS camera 25 and a cell to be detected 26.

Laser beams emitted by a continuous working laser 1 in the system are coupled into a two-port optical fiber beam splitter 3 through an output coupling optical fiber 2 and are respectively coupled into single-mode optical fibers 4 and 5 according to a certain intensity ratio. The laser beam transmitted by the single mode fiber 5 is divided into two beams of laser with equal intensity by the two-port fiber beam splitter 6, and the two beams of laser are transmitted by the single mode fibers 7 and 8, coupled by the fiber beam combiner 9, and processed into the double-core fiber 10 with the output end being processed into the frustum with a specific angle, so that the stable capture of the cell to be detected is realized. The laser beam passing through the single mode fiber 4 is frequency modulated by the fiber frequency modulator 11 to generate a laser pulse with adjustable pulse width, and then is split into two beams of laser with equal intensity by the two-port fiber beam splitter 12, and the two beams of laser are respectively coupled into the

single mode fibers

13 and 14. The laser pulses coupled into the single mode fiber 13 are pulse intensity modulated via a fiber intensity modulator 16. The laser pulses coupled into the single mode fiber 14 are pulse intensity modulated by a fiber intensity modulator 16 and a time delay is introduced by a fiber time delay 17. Two laser pulses with certain time delay are coupled into an output end through an optical fiber beam combiner 18 and processed into a double-core optical fiber 19 of a frustum with a specific angle, so that accurate control of the space angle of the cell to be detected around the capture axis is realized. The white light emitted by the white light LED light source 20 is reflected by the 45-degree reflector 21, and then forms a full-field uniform kohler illumination on the sample 26 to be measured by the condenser 22. The image information of the sample 26 to be measured is collected by the apochromatic microscope objective 23, the background noise is eliminated by the optical filter 24, and the spatial position information of the cell is recorded by the CMOS camera 25. The invention can be used for realizing stable and accurate capture of specific living body unicells and accurate control of space angles, and can be widely used for long-time observation and research of the life activity process of the specific living body unicells.

Two double-core optical fibers which are oppositely arranged, wherein laser emitted by the continuous working laser 1 is divided into two beams with different intensity proportions by the two-port beam splitter 3, wherein the intensity of a light beam entering the single-mode optical fiber 5 is smaller than that of a light beam entering the single-mode optical fiber 4, and the light intensity for capturing cells is larger than that of light for controlling the cells, so that the cells are prevented from being separated from a control range due to insufficient captured light intensity when the cells are controlled. The light beam in the single mode fiber 5 is split into two beams by a beam splitter 6, and is input into a double-core fiber 10 by a fiber combiner 9. The light is used for stably capturing the cells and fixing the cells. Light in the single-mode fiber 4 is modulated by the frequency modulator 11 to become pulse light, the pulse light is divided into two beams by the beam splitter and finally output by the double-core fiber 19, and the light is used for controlling cells. The optical fiber 13 and the optical fiber 14 each have an

intensity modulator

The above examples are provided for the purpose of describing the invention only, and are not intended to limit the scope of the invention. The scope of the invention is defined by the appended claims. Various equivalent substitutions and modifications can be made without departing from the spirit and principles of the invention, and are intended to be within the scope of the invention.

Claims

1. A method and device for precise active light manipulation based on double-core optical fiber in the rotation of a living single cell. It is characterized in that the device is mainly composed of two double-core optical fibers 10 and 19 whose output end faces are processed into a specific angle, the central axes of the fiber cores are perpendicular to each other, and the output end faces are fixed and installed facing each other. CW laser 1, output coupling fiber 2, two-port fiber splitter 3, 6, 12, single-mode fiber 4, 5, 7, 8, 13, 14, fiber combiner 9, 18, fiber frequency modulator 11 , fiber intensity modulators 15, 16, fiber time delay 17, dual-core fibers 10, 19 whose output ends are processed into a frustum of a specific angle, white LED light source 20, total reflection mirror 21, condenser 22, apochromatic microscope objective 23, an optical filter 24, a CMOS camera 25 and a living single cell 26 are composed.

In the system, the laser beam output by the continuous working laser 1 is coupled into the two-port fiber beam splitter 3 through the output coupling fiber 2, and is respectively coupled into the single-mode fibers 4 and 5 according to the intensity ratio of 7/3. After the laser beam with the intensity of 70% is transmitted through the single-mode fiber 5, it is divided into two laser beams with the same intensity through the two-port fiber beam splitter 6, and then transmitted through the single-mode fibers 7 and 8, and then coupled into the output through the fiber combiner 9 The two cores of the twin-core optical fiber 10 whose ends are machined into a frustum of a specific angle. The central axes of the two cores of the dual-core fiber 10 are fixed horizontally in a culture dish containing living cells, and the light field formed by the laser beam output from the output end acts on the living single cells to achieve stable capture of them.

After the laser beam with an intensity of 30% is transmitted through the single-mode fiber 4, the frequency is modulated by the fiber frequency modulator 11 to generate a laser pulse with adjustable pulse width. The laser pulse is divided into two laser beams with the same intensity through the two-port fiber beam splitter 12, which are respectively coupled into the single-mode fibers 13 and 14. The laser pulses transmitted through the single-mode fibers 13 and 14 are subjected to pulse intensity modulation by the fiber intensity modulators 15 and 16 . A fiber time delay 17 is used to introduce a certain time delay between the laser pulses transmitted via the single mode fibers 13 , 14 . Two laser pulses with a certain time delay are coupled into the two cores of the double-core optical fiber 19 whose output end is processed into a frustum of a specific angle through the fiber combiner 18 . The central axes of the two cores of the double-core optical fiber 19 are vertically fixed in the culture dish containing the living cells 26 to be tested, and the laser beam output from the output end acts on both ends of the cells to be tested, realizing the spatial rotation of the living single cells around the capture axis Precise active light manipulation of angles.

The white light emitted by the white light LED light source 20 is reflected by a 45-degree total reflection mirror 21 , and then passes through a condenser lens 22 to form a uniform Kohler illumination in the entire field on the sample to be tested 26 . The image information of the living single cell 26 is collected by the apochromatic microscope objective lens 23 , the background noise is eliminated by the filter 24 , and the spatial position information of the cell is recorded by the CMOS camera 25 . The invention can be used to achieve stable and precise capture of specific living single cells and precise active light manipulation of spatial rotation angles, and can be widely used for long-term observation and research on the physical and chemical properties of specific living single cells' life activities.

2 . The light manipulation system according to claim 1 : it is composed of two double-core optical fibers 10 and 19 whose output end faces are processed into a truncated cone with a specific angle. 3 . It is characterized in that: the optical manipulation device designed by the present invention is a double-core optical fiber 10, 19 processed into a cone frustum of a specific angle through two output end faces, the central axes of the cores are perpendicular to each other, and the output end faces are fixed and installed facing each other. The laser beam output by the continuous working laser 1 is divided into two beams according to the intensity ratio of 7/3. One of the laser beams with high intensity is transmitted through the two cores of the dual-core optical fiber 10 fixedly installed on the center axis of the core, and then processed. A Bessel light field distribution is formed in the horizontal plane of the output end of the truncated cone with a specific angle, and another laser beam with low intensity is modulated by the fiber frequency modulator 11 to generate a laser pulse with adjustable pulse width. The laser pulse is divided into two beams of equal intensity by the optical fiber beam splitter 12, and the intensity is modulated by the optical fiber intensity modulators 15 and 16 respectively. Two laser pulses. After the two laser pulses are transmitted through the two cores of the twin-core optical fiber 19 fixedly installed with the central axis of the core perpendicular to the horizontal plane, a light field is formed in the vertical plane of the output end of the frustum processed into a specific angle. Due to the different angles of the frustum at the output end of the two dual-core fibers, the focused position of the focused laser light field at the output end is different. The laser beam transmitted through the double-core optical fiber 10 acts on a specific living single cell at the output end to realize its stable capture. The laser pulses transmitted through the double-core optical fiber 19 act on both ends of the captured living single cell at the output end, respectively. When a laser pulse output from a fiber core acts on one end of the cell, during the pulse duration period, the laser pulse continues to exert a light driving force on one end of the cell, pushing the cell to rotate around the rotational axis formed by the captured light field. The laser pulses applied to the other end of the cell arrive after the cell is rotated by a certain angle under the action of the photopropulsive force. During the duration of the laser pulse, the photobraking force applied in the opposite direction and the resistance received during the cell rotation process work together to achieve precise active optical manipulation of the cell rotation angle.