TW202022353A - Light-trapping microorganism system and method thereof - Google Patents

Abstract

A light-trapping microorganism system includes a microorganism container, a microscope device and an optical tweezer. The microorganism container contains a microorganism, with the microorganism container being aligned with the microscope device for displaying the microorganism. The optical tweezer generates a predetermined light beam to trap the microorganism thereon. The optical tweezer and the trapped microorganism are moved together by a predetermined speed from a first position to a second position, with displaying the optical tweezer and the trapped microorganism via the microscope device.

Description

Light capture microorganism system and method

The present invention relates to a light-trapping microbe (microorganism) system and its method; in particular, it relates to a laser beam (laser beam) microbial trapping system and its method; more particularly to an optical tweezer ] System and method for capturing microorganisms; more particularly, it relates to a system and method for capturing microorganisms with optical fiber (optical fiber); more particularly, it relates to a system and method for capturing microorganisms with laser optical fiber optical tweezers.

For example, regarding the conventional manufacturing method of optical fiber lens [fiber microlens], for example, the invention patent of "Method of Manufacturing Hyperbolic Optical Fiber Lens" in the Republic of China Patent Publication No. TW-I241421, which discloses a manufacturing method of hyperbolic optical fiber lens的方法。 The method. The manufacturing method of the optical fiber lens includes the following steps: [a] stripping off a predetermined length of cladding layer of an optical fiber to form an exposed part; [b] cleaning the exposed part; [c] fixing the optical fiber to be processed to an optical fiber In the fixed seat; [d] Provide a container including a hydrofluoric acid layer, an oil layer and an intermediate mixing layer; [e] Extend the bare part of the optical fiber into the hydrofluoric acid layer to be formed by etching A cone angle; [f] use two arcs to melt the cone angle; and [g] adjust the relative position of the two arcs and the cone angle, and use the uneven temperature field formed by the two arc discharges to obtain a hyperbolic optical fiber The lens and the set radius of curvature improve the coupling efficiency of the optical fiber.

However, the aforementioned No. TW-I241421 only discloses the manufacturing method of the fiber lens and its technology suitable for mass production. In addition, the fiber The lens manufacturing method also only reveals how to form a perfect match between the fiber lens and the laser light field, and adjust the relative position of the two arcs and the cone angle of the fiber lens to obtain the best radius of curvature of the fiber lens, and to couple it. Light efficiency has a decisive influence on technology.

Another conventional fiber optic tweezers [fiber optic tweezers] manufacturing method, for example, the Republic of China Patent Publication No. TW-I474061 "The manufacturing process of fiber optic tweezers" invention patent, which discloses a manufacturing method of optical tweezers. The optical tweezers manufacturing method includes the following steps: [a] Stripping: cutting an optical fiber of an appropriate length, and stripping off one of the coating layers (coating) of the optical fiber to leak a bare optical fiber, which includes a fiber coating layer (cladding) ] And a core layer [core]; [b] cleaning: clean the optical fiber; [c] cutting: cutting and flattening the end face of the bare optical fiber; [d] etching: fixing the optical fiber in a container Add a buffered oxide etch solution [buffered oxide etch, BOE], set the fixed optical fiber corresponding to the container, immerse the end of the optical fiber to be processed into the oxide buffered etching solution for etching, and the end of the optical fiber can be formed A tapered shape to make a fiber microlens or a lensed fiber [fiber microlens]; you can also choose to further melt the tapered fiber end with an arc device at both ends to form a hemispherical fiber microlens.

In addition, the aforementioned No. TW-I474061 utilizes the hemispherical lensed optical fiber of the optical fiber optic tweezers to achieve a simpler overall production and lower production cost. If it uses light or light energy to capture tiny objects or microorganisms in a non-contact manner in the field of biotechnology or biomedical applications, and it has high capturing efficiency and low capturing power.

However, the aforementioned No. TW-I474061 only discloses that the fiber optic tweezers are suitable for non-contact capture of small objects or microorganisms. In addition, the fiber optic tweezers are suitable for non-contact capture of small objects or microorganisms, and there must be a potential demand for future applications of other biotechnologies (for example, the application of microorganism identification).

Another conventional fiber optic tweezers [fiber optic tweezers] manufacturing method, for example: US Patent No. US-9958663 "light-trapping cancer cell stage testing method" invention patent, which discloses a light-trapping cancer cell stage identification method . The light-captured cancer cell stage identification method includes: measuring the average escape speed or range of a first cancer cell and the average escape speed or range of a second cancer cell, and the cancer cells of the first cancer cell and the second cancer cell The stages are known to be different, and the first cancer cell and the second cancer cell are of a known type; then the average escape speed of the first cancer cell and the average escape speed of the second cancer cell are used to calculate an escape speed ratio or its range; Select a cancer cell to be identified, and measure the escape speed of a cancer cell to be identified, and the cancer cell to be identified is of a known type but unknown stage; use the escape speed ratio or its range to compare the escape speed of the cancer cell to be identified ; And according to the comparison result of the escape speed of the cancer cells to be identified, a cancer cell stage is determined.

However, the aforementioned No. US-9958663 only discloses a method for identifying cancer cell stage by light capture, and the method for identifying cancer cell stage is not suitable for identifying other objects. In addition, the light-captured cancer cell stage identification method is suitable for non-contact capture of small objects or microorganisms, and there must be a potential demand for future applications of other biotechnologies (such as applications for identification of various types of microorganisms).

The aforementioned ROC Patent Publication Nos. TW-I241421, TW-I474061, and US-9958663 for invention patents are only a reference for the technical background of the present invention and illustrate the current state of the technology development. They are not intended to limit the scope of the present invention. .

In view of this, in order to meet the above technical problems and needs, the present invention provides a light-trapping microorganism system and method, which uses an optical fiber optical tweezers to capture a microorganism (a known species of microorganisms), so that the optical fiber optical tweezers A captured microorganism is formed on the top, and the fiber optic tweezers and the captured microorganism are controlled to move at a predetermined speed to measure a microorganism's escape speed (or speed range or average escape speed), and optional Choose to establish a microbial identification standard database, and then use the microbial escape speed (or speed range or average escape speed) to compare with the escape speed of a microorganism to be identified (unknown species of microorganism), so as to compare the escape speed of the microorganism to be identified The result is to determine a microorganism type to expand the further application range of conventional fiber optic tweezers.

The main purpose of the present invention is to provide a light trapping microorganism system and method, which uses an optical fiber optical tweezers to capture a microorganism with a predetermined light beam, so as to form a trapped microorganism on the optical fiber optical tweezers, and combine the The optical fiber optical tweezers and the captured microorganisms move at a predetermined speed, and the optical fiber optical tweezers and the captured microorganisms are displayed through the microscope device, and the optical fiber optical tweezers and the captured microorganisms are moved from a first position Move to a second position to achieve the purpose of accurately capturing microorganisms.

Another object of the present invention is to provide a method for identifying microbes captured by light, which uses an optical fiber optical tweezers to capture a microorganism (known species of microorganisms) so as to form a captured microorganism on the optical fiber optical tweezers, and another Control the fiber optic tweezers and the captured microorganisms to move at a predetermined speed to measure a microorganism escape speed (or speed range or average escape speed), and optionally establish a microorganism identification standard database, and then use the microorganism to escape The speed (or speed range or average escape speed) is compared with the escape speed of a microorganism to be identified (unknown species of microorganism), so that a microorganism type is determined based on the comparison result of the escape speed of the microorganism to be identified, so as to achieve accurate identification of the microorganism type purpose.

In order to achieve the above objective, the light-trapped microorganism identification method of a preferred embodiment of the present invention includes: using an optical fiber optical tweezers to capture a microorganism (known species of microorganisms) so as to form a captured microorganism on the optical fiber optical tweezers ; Control the fiber optic tweezers and captured microorganisms to a pre- Move at a constant speed to measure the escape speed of a microorganism (or speed range or average escape speed); compare the escape speed of the microorganism (or speed range or average escape speed) with the escape speed of a microorganism to be identified (unknown species of microorganism) ; And determine a microorganism type according to the comparison result of the escape speed of the microorganism to be identified.

The preferred embodiment of the present invention selects various microorganisms as a reference microorganism species category.

The preferred embodiment of the present invention establishes a microbial identification standard database required by the light capture test method.

The optical fiber optical tweezers of the preferred embodiment of the present invention are used to measure a capturing force, a capturing efficiency or a combination thereof.

In a preferred embodiment of the present invention, the microorganism is placed in a buffer.

In the preferred embodiment of the present invention, the predetermined speed is a fixed speed.

In the preferred embodiment of the present invention, the microorganism escape speed is a range of microorganism escape speed or an average microorganism escape speed.

In the preferred embodiment of the present invention, the microbial escape speed range is between a maximum microbial escape speed and a minimum microbial escape speed.

The escape speed of the microorganism to be identified in the preferred embodiment of the present invention is selected from the result of measuring the escape speed of five times and calculating the average.

The microorganism in the preferred embodiment of the present invention is selected from a bacterium, a fungus, a virus, an algae or a pathogenic bacterium.

In a preferred embodiment of the present invention, the microorganism is selected from a yeast (for example: natural yeast, steamed bread yeast, beer yeast or wine yeast), a probiotic or a pathogenic bacteria.

The optical fiber optical tweezers in the preferred embodiment of the present invention is a laser optical fiber optical tweezers.

The optical fiber optical tweezers fixture of the preferred embodiment of the present invention has a capturing inclination or a working inclination, and the capturing inclination or the working inclination is 40° or 50°.

The optical fiber optical tweezers holder of the preferred embodiment of the present invention has a single-mode/multi-mode fiber or a single-mode/multi-mode fiber micro lens.

In order to achieve the above-mentioned object, the light-trapping microorganism system of a preferred embodiment of the present invention includes: a microorganism container for accommodating a microorganism; at least one microscope device corresponding to the microorganism container so that the microscope device can display the microorganism Microorganisms of the microbial container; and at least one optical fiber optical tweezers, which uses the optical fiber optical tweezers to capture at least one microbial body with a predetermined light beam in the microbial container, so as to form a captured microorganism on the optical fiber optical tweezers ; Wherein the optical fiber optical tweezers and the captured microorganisms are controlled to move at at least a predetermined speed, and the optical fiber optical tweezers and the captured microorganisms are displayed through the microscope device, and the optical fiber optical tweezers and the captured microorganisms are displayed Move from a first position (for example: inside the microorganism container) to a second position (for example: other positions).

In order to achieve the above objective, the method for capturing microorganisms by light in a preferred embodiment of the present invention includes: providing a microorganism container, and the microorganism container is used for accommodating a microorganism; providing at least one microscope device, and the microscope device corresponds to the microorganism container , In order to use the microscope device to display the microorganisms of the microorganism container; provide at least one optical fiber optical tweezers, and use the optical fiber optical tweezers to capture at least one microorganism in the microorganism container with a predetermined light beam, In order to form a captured microorganism on the optical fiber optical tweezers; control the optical fiber optical tweezers and the captured microorganisms to move at at least a predetermined speed, and display the optical fiber optical tweezers and the captured microorganisms through the microscope device , And move the fiber optic tweezers and the captured microorganisms from a first position (for example: inside the microorganism container) to a second position (for example: other positions).

The microscope device of the preferred embodiment of the present invention is connected to at least one image capturing system, and the microscope device corresponds to the microorganism, so that the image capturing system captures a microorganism image from the microscope device.

The preferred embodiment of the present invention further includes a work platform and a translation platform control device, and the microorganism container is arranged on the work platform, and the translation platform control device is used to control the work platform.

The preferred embodiment of the present invention further includes a computer system, and the computer system is connected to the microscope device, or the computer system is connected to a laser light source device, or the computer system is connected to a work platform, a translation stage control device, or An image capture system.

The optical fiber optical tweezers of the preferred embodiment of the present invention are arranged in a laser light source device.

In a preferred embodiment of the present invention, the microorganism is selected from a yeast (for example: natural yeast, steamed bread yeast, beer yeast or wine yeast), a probiotic or a pathogenic bacteria.

1‧‧‧Computer system

2‧‧‧Image capture system

21‧‧‧Microscope device

22‧‧‧Filter

3‧‧‧Laser light source device

31‧‧‧Laser light source

32‧‧‧Fiber Optic Coupler

33‧‧‧Optical fiber

34‧‧‧Fiber Microlens

5‧‧‧Working platform

50‧‧‧Microbial culture container to be identified

51‧‧‧Pan stage control device

52‧‧‧The first electronic control platform

53‧‧‧ Stage

54‧‧‧Light source of stage

55‧‧‧Second Electronic Control Platform

9‧‧‧Microorganism

Figure 1: A schematic diagram of a light trapping microbe system and its optical tweezers operating system according to a preferred embodiment of the present invention.

Figure 2: A schematic diagram of the optical tweezers operating system of the preferred embodiment of the present invention performing light trapping of microorganisms.

Figure 3: A schematic flow diagram of a light-trapping microorganism identification method according to a preferred embodiment of the present invention.

Figure 3A: A schematic diagram of the establishment process of the light-captured microorganism identification standard database according to a preferred embodiment of the present invention.

Figure 3B: A schematic flow diagram of a method for identifying microorganisms by light trapping in a preferred embodiment of the present invention.

Figure 4: A schematic diagram of the mechanism of light refraction generated by the light beam under the force of the microorganisms when the light trapping microorganisms system and the method of the preferred embodiment of the present invention begin to capture microorganisms.

Figure 5: When the light trapping microorganism system and method of the preferred embodiment of the present invention begin to capture microorganisms, the microorganisms are subjected to the reaction force of the light beam to generate light trapping mechanism.

Figure 6: The light trapping microorganism system and method of the preferred embodiment of the present invention is a schematic diagram of the phenomenon that the microorganisms have been stabilized by the light beam when the microorganisms have been captured.

Fig. 7A: A schematic diagram of a microscopic image of the light-trapping microorganism system and its method for measuring the first yeast in a preferred embodiment of the present invention.

Fig. 7B: A schematic diagram of a microscopic image of the light-trapping microorganism system and its method for measuring the second yeast in a preferred embodiment of the present invention.

Fig. 7C: A schematic diagram of a microscopic image of the light-trapping microorganism system and its method for measuring the third yeast in a preferred embodiment of the present invention.

Figure 8: A schematic diagram of the light trapping microorganism identification method of the preferred embodiment of the present invention used to measure the first, second and third yeasts to obtain their escape speed.

Figure 9: A schematic diagram of the light trapping microorganism identification method of the preferred embodiment of the present invention used to measure the first, second and third yeasts to obtain their trapping efficiency.

In order to fully understand the present invention, preferred embodiments are exemplified below and described in detail with the accompanying drawings, and they are not intended to limit the present invention.

The light trapping microorganism system of the preferred embodiment of the present invention and the same The method or light-trapping microorganism identification method can be applied to capture or identify various types of microorganisms (bacteria are prokaryotes, fungi, algae, protozoa are eukaryotes, viruses have no cell structure), such as bacteria, pathogenic bacteria (e.g. : Salmonella, Vibrio enteritidis, Vibrio bacillus, Escherichia coli, Clostridium botulinum or Staphylococcus], probiotic cells (e.g. natural yeast, steamed bread yeast, beer yeast or wine yeast) or other microbial cells, but It is not intended to limit the scope of application of the present invention; furthermore, the light trapping microorganism system and its method or the light trapping microorganism identification method of the preferred embodiment of the present invention can use light of various suitable wavelength ranges, for example: laser light of various wavelengths [Infrared laser, red laser, green laser, blue laser or purple laser, etc.], but it is not intended to limit the scope of application of the present invention.

The optical trapping microorganism system and its method or the optical trapping microorganism identification method of the preferred embodiment of the present invention select the preset optical fiber optical tweezers system parameters. It uses a fiber optic tweezer to measure the escape velocity and diameter of several known types of microorganisms. It uses statistical mathematics methods to calculate the average escape velocity and escape velocity range of each known species of microorganisms, and selects various reference species of microorganisms to establish a microbial identification standard database required by the light capture inspection method. Then use the same fiber optic tweezers system parameters and experimental methods to measure the escape velocity and diameter of a microorganism to be identified, and calculate the average value of the escape velocity ratio and the capture efficiency. The ratio of the escape rate of the microorganism to be identified and the capture efficiency is compared with the value of the same type of microorganism in the standard database, so that a microorganism type is determined based on the comparison result of the ratio of the escape rate of the microorganism to be identified to the capture efficiency.

Figure 1 shows a schematic diagram of the light trapping microbe system and its optical tweezers operating system according to a preferred embodiment of the present invention. Please refer to Figure 1. For example, the preferred embodiment of the present invention uses optical tweezers in terms of structure. The operating system includes a computer system 1, an image capturing system 2, a microscope device 21, and a laser light source device. 3. A working platform 5 and a translation stage control device 51, which are appropriately connected to each other and assembled or correspondingly configured to form the Optical tweezers operating system.

Please refer to Figure 1 again. For example, the computer system 1 chooses to include a computer host (computer device) and a display device (display device) or other peripheral devices (peripheral), and the computer system 1 is appropriately configured At a predetermined location, and the computer host of the computer system 1 is connected to the image capturing system 2 or the microscope device 21. In addition, the computer system 1 can also be connected to the laser light source device 3, the work platform 5, or the translation stage control device 51.

Please refer to Figure 1 again. For example, the image capturing system 2 has an image capturing processing unit, and the image capturing system 2 is connected and combined with the microscope device 21, or the image capturing system 2 can be Choose to connect any electron microscope device to capture at least one microscopic image (such as microbial cell image) or a series of electron microscopic images from the microscope device 21. In addition, the image capturing system 2 includes a filter 22 or other equipment.

Please refer to Figure 1 again. For example, the microscope device 21 may optionally include a CCD device [Charge Coupled Device] and a microscope or other functional devices (such as a fixing device), and the CCD device is combined on the microscope In order to use the CCD device to capture at least one microscopic image or a series of microscopic images through the microscope. During the assembling operation, the microscope device 21 corresponds to the work platform 5 so as to capture images of the work in the area above the work platform 5.

Please refer to Figure 1 again. For example, the laser light source device 3 is selected from a predetermined wavelength laser light source device (for example: 650nm or other wavelengths), and the laser light source device 3 optionally includes a laser light source [Laser source, such as semiconductor laser or other laser] 31, a fiber coupling device 32, an optical fiber 33 and a fiber micro lens 34, and the laser light source 31 passes through the fiber coupling device 32 is connected to the optical fiber 33, and the optical fiber 33 and its optical fiber microlens 34 are used to properly emit a laser Beam [laser beam] to form a fiber optic tweezers. In the identification of microorganisms, the optical fiber optical tweezers are suitable for light capture of microorganisms. In the preferred embodiment of the present invention, the optical fiber 33 of the optical fiber optical tweezers has a single-mode/multimode optical fiber, and the optical fiber microlens 34 of the optical fiber optical tweezers has a single-mode/multimode optical fiber microlens.

Please refer to Figure 1 again. For example, the working platform [operational platform] 5 includes a translation stage control device 51, a first electronic control platform [XY axis electric control platform] 52, and a stage 53, A stage light source 54 and a second electronic control platform [XYZ axis electronic control platform] 55 or other automatic electronic control tabletops, so that the working platform 5 can be operated automatically or electronically. Place a microorganism culture container or a device with similar functions on the stage 53 of the work platform 5, and use the first electronic control platform 52 of the work platform 5 to properly adjust and operate the stage 53 and the stage The horizontal position of the light source 54 and the second electronic control platform 55 of the working platform 5 are used to properly adjust and operate the [capture] working inclination angle, working distance, horizontal position and vertical position of the optical fiber 33 and the optical fiber microlens 34. In addition, there is a predetermined illumination angle between the stage 53 and the stage light source 54.

Figure 2 shows a schematic diagram of the optical tweezers operating system of the preferred embodiment of the present invention for light trapping microbes. Please refer to Figure 2. For example, the optical fiber 33 of the optical tweezers operating system of the preferred embodiment of the present invention selects an optical fiber with a diameter of 125μm, and the core diameter of the unetched section is 8μm, which has been etched The fiber diameter of the section is about 14μm, and the fiber microlens 34 is located at the end of the etched section, and the fiber microlens 34 is a cone, and it has a tapered angle (tapered angle) α, which is between Cone angle range between 90° and 115° or other angles.

Please refer to Figures 1 and 2. For example, the laser light source device 3 chooses to adjust the capture power to 7mW or other appropriate capture power (for example: 5mW, 20mW or its interval), and the optical fiber etched cone 34 Corresponding to a microorganism culture container 50 to be identified, with light [laser] Capture a microorganism [microorganism to be identified] 9. At this time, the operator can choose to use the first electronic control platform 52 of the work platform 5 to appropriately adjust the angle and horizontal position of the stage 53 and the microorganism culture container 50 to be identified. The microorganism 9 in the preferred embodiment of the present invention is placed in a buffer solution.

Please refer to Fig. 2 again. For example, when the microorganism culture container 50 to be identified is performing light-trapping microorganism identification operation, it can be operated to the tip of the end surface of the optical fiber micro lens 34 of the laser light source device 3 and the microorganism There is a predetermined working distance (working distance) D or a predetermined working distance range between the bodies 9 and a predetermined working inclination (working distance) between the longitudinal axis of the optical fiber microlens 34 of the laser light source device 3 and the microorganism 9 angle]θ or a predetermined working inclination range, that is, the angle between the beam's traveling direction and the horizontal direction.

Figure 3 shows a schematic flow diagram of a light-trapping microorganism identification method according to a preferred embodiment of the present invention. Please refer to Figures 1, 2 and 3, for example, the light-trapping microorganism identification method of the preferred embodiment of the present invention includes the first step S1: First, use the optical tweezers operating system or the optical fiber optical tweezers to properly capture Measure a microorganism (a known type of microorganism) to form a captured microorganism on the fiber optic tweezers. The microorganism of the preferred embodiment of the present invention can also be selected from a bacterium, a fungus, a virus, an algae, a protozoan (protozoa) or a pathogenic bacteria.

Please refer to Figures 1, 2 and 3 again. For example, the light-trapping microorganism identification method of the preferred embodiment of the present invention includes a second step S2: Next, the optical fiber optical tweezers and the captured microorganisms are controlled to at least A predetermined speed and appropriate technical means (for example: dynamic measurement method [dynamic measurement method] or static measurement method [static measurement method] move to measure the escape velocity of a microorganism [or velocity range or average escape velocity]. Use the optical tweezers operating system or optical fiber optical tweezers to select and measure one of the first microorganisms (such as beer yeast) The average escape velocity of the first microorganism or its range and the average escape velocity of a second microorganism (for example: Saccharomyces cerevisiae) or its range.

Another preferred embodiment of the present invention establishes a microbial identification standard database required by the light capture test method to store the known average escape velocity or range of the first microorganism and the average escape velocity or range of the second microorganism, which can also be used Store the known microbial diameter. The average escape velocity of the first microorganism (or the average escape velocity of the second microorganism) is between a maximum average escape velocity of microorganisms and a minimum average escape velocity of microorganisms.

Please refer to Figures 1 and 2. For example, the preferred embodiment of the present invention selects various microorganisms as a reference microorganism species, and the optical fiber tweezers are used to measure a capturing force (or lateral capturing force, transverse trapping force], a trapping efficiency, or a combination thereof, and the predetermined speed can be selected as a fixed speed (ie, uniform speed), as shown in the y-axis direction in Figure 2.

Please refer to Figures 1 and 2. For example, the light-trapping microorganism identification method of the preferred embodiment of the present invention includes the third step S3: Then, the microorganism escape speed (or speed range or average escape speed) is used with The escape speed of a microorganism to be identified (an unknown species of microorganism) is compared, and a microorganism to be identified (or a microorganism to be tested) is selected, and the fiber optic tweezers can be used to measure the escape speed of a microorganism to be identified automatically or in other ways. The escape speed of the microorganism to be identified in the preferred embodiment of the present invention is selected from the result of measuring the escape speed of five times and calculating the average.

Please refer to Figures 1 and 2. For example, the light-captured microorganism identification method of the preferred embodiment of the present invention includes a fourth step S4: Next, identify and determine a microorganism based on the comparison result of the escape speed of the microorganism to be identified species. The escape speed (or speed range or average escape speed) of the microorganisms can be compared with the escape speed of the microorganisms to be identified in automatic or other ways. In the comparison of microbial identification, the light-captured microbial identification method of the preferred embodiment of the present invention and its establishment of a microbial identification standard database can be Choose to use mathematical formulas or formulas for identifying various types of microorganisms.

FIG. 3A shows a schematic diagram of the establishment process of the light-trapping microorganism identification standard database according to a preferred embodiment of the present invention. Please refer to Fig. 3A. For example, the establishment of the light-trapping microorganism identification standard database of the preferred embodiment of the present invention includes: setting the parameters of the optical tweezers system; selecting several known types of yeast (or other microorganisms) Measure the diameter of various yeast cells (or other microorganisms) and their escape speed several times; calculate the average value of the escape speed of various yeast cells (or other microorganisms); establish the diameter of various yeast cells (or other microorganisms), Database of average escape speed and range of escape speed; standard database for identification of light-trapped yeast (or other microorganisms) required to complete the classification of live bacteria.

Figure 3B shows a schematic flow diagram of the light-trapping microorganism identification method according to a preferred embodiment of the present invention. Please refer to Figure 3B. For example, the light-trapping microorganism identification method of the preferred embodiment of the present invention includes: setting the optical tweezers system parameters according to the standard database, and setting the same experimental parameters according to the standard database; selecting unknown species Number of yeasts (or other microorganisms); measure the diameter of various yeasts (or other microorganisms) and their escape speed several times; measure the diameter, average escape speed and capture efficiency of each yeast (or other microorganisms) to be tested; According to the established standard database, complete the identification of each yeast (or other microorganism) type to be tested.

Please refer to Figures 1, 2 and 3A again. For example, the preferred embodiment of the present invention uses various mathematical formulas or formulas for the identification of microorganisms in the establishment of a standard database for identification of light-capture microorganisms, and selects one The mathematical formula is illustrated below.

For example, the light trapping microorganism identification method of the preferred embodiment of the present invention adopts the trapping force F as: F = 6 πηrv

Where η is the viscosity coefficient of the solution, r is the radius of the sample (similar to a sphere), and v is the escape velocity of the sample.

For example, the optical capture microorganism identification method of the preferred embodiment of the present invention adopts the capture efficiency Q as:

Where c is the speed of light in vacuum, n is the refractive index of the solution, and P is the capture power of the optical tweezers.

Please refer to Figures 1 and 2 again. For example, if the output optical power of the same fiber increases, the escape speed required by the captured object will also increase. Therefore, when the output light power is fixed, each type of microorganism has its corresponding escape speed range, so that the identification test of the microorganism type can be achieved. Generally speaking, under the condition of the same light capture power, the escape speed of microorganisms of the same type but with different diameters is not obvious; while the difference in escape speed between different types of microorganisms is relatively obvious. As for the escape speed of microorganisms, their various optical properties (refraction, reflection, transmission, absorption) are different depending on the internal biochemical composition of their organisms, which further results in different escape speeds.

Generally speaking, the capturing principle of single-beam optical tweezers can be explained by geometric optics [ray optics]. When light travels through media with different refractive indices, the difference in the speed of light will cause the direction of light to bend, and cause light refraction (refraction) phenomena and light momentum (momentum) changes. The principle of optical tweezers is to use the principle or mechanism of light refraction and the change of light momentum to optically capture microorganisms (transparent and tiny objects). That is, when the laser light (light field) approaches or enters the microorganisms to be captured, the light refractions in the microorganisms, and changes the light momentum to capture the microorganisms. If you encounter opaque microorganisms, you need to use double-beam optical tweezers to capture. The capture principle is explained by the theory of radiation pressure or light pressure (radiation pressure).

Figure 4 discloses the light trapping microorganisms of the preferred embodiment of the present invention When the system and method begin to capture microorganisms, the light beam is subjected to the force of the microorganisms to produce a schematic diagram of the light refraction mechanism. Please refer to FIGS. 1, 2 and 4. For example, the light-trapping microorganism identification method of the preferred embodiment of the present invention uses the light beam output by the optical fiber microlens 34 to capture a microorganism 9. The optical fiber optical tweezers clamp is a laser optical fiber optical tweezers clamp (for example: 650nm laser optical fiber optical tweezers clamp). The optical fiber optical tweezers clamp has a capturing inclination or a working inclination, and the capturing inclination or the working inclination is 40°, 50° or other appropriate angles. The fiber optic tweezers holder has a single-mode fiber or other appropriate fiber, and the fiber is processed into a fiber micro lens.

Please refer to Figure 4 again. When the light beam (laser beam) passing through the optical fiber microlens is incident on the surface of the microorganism 9, if it is not deflected, the penetrating light should travel a straight path (two symmetrical dotted lines) However, because the microbial body’s refractive index (refractive index) is different from the surrounding environment, the incident light entering the microbial body is deflected (shown by the two symmetrical solid lines, that is, the refracted light), and the internal material of the microorganism 9 The two sides of the center of symmetry are deflected inward [to the longitudinal axis of the optical fiber microlens 34], as if the internal substance of the microorganism exerts a force F1 [indicated by the two symmetrical inward arrows] on the penetrating light, causing it to deviate The path shown by the original dashed line is the change of light momentum generated by the refracted light compared with the original incident light.

Figure 5 shows a schematic diagram of the light trapping mechanism generated by the light trapping mechanism when the microorganisms are subjected to the reaction force of the light beam when the light trapping microorganism system and the method of the preferred embodiment of the present invention begin to trap microorganisms. Please refer to Figures 4 and 5, the internal matter of the microorganism 9 exerts a force F1 (shown by the two symmetrical inward arrows in Figure 4) on the incident light, which deflects the incident light, according to Newton's third According to the law of motion, the refracted light also produces a force F2 of the same magnitude but opposite directions (shown by the two symmetrical outward arrows in Figure 5) on the internal matter of the microorganism 9, namely photons (refracted light). The reaction force F2 is on the microorganism 9. The microbial body 9 is subjected to the sum of the vertical vectors F3 of the reaction force F2 (shown by the vertical upward arrow in Figure 5). Proper movement is generated along the longitudinal axis of the fiber microlens 34.

Fig. 6 shows a schematic diagram of the phenomenon that the light trapping microorganism system and method of the preferred embodiment of the present invention have been trapped by the light beam when the microorganisms have been captured. Please refer to Figures 5 and 6, when a photon [refracted light] simultaneously exerts a reaction force F2 on the microorganism 9, the microorganism 9 is subjected to the sum of the vertical vectors of the reaction force F2 (as shown by the vertical upward arrow in Figure 5). As shown], proper movement can be generated along the longitudinal axis of the optical fiber microlens 34. When the microorganism 9 moves to the optical focus of the fiber microlens 34, the [laser] beam incident on the fiber microlens 34 generates a stable optical potential well [optical potential well] near the focus, and The potential energy well forms a force balance point (a position where the resultant force is zero), and forms a trap to restrict the microorganism 9. At this time, once the optical fiber microlens 34 clamped by the optical tweezers is moved, the microorganism 9 also moves along with it, and the light beam of the optical fiber microlens 34 can be used to control the movement of the microorganism 9.

FIG. 7A shows a schematic diagram of a microscopic image of the light capturing microorganism system and its method for measuring the first yeast in a preferred embodiment of the present invention. Please refer to Figure 7A, the light-trapping microorganism system and method and the light-trapping microorganism identification method of the preferred embodiment of the present invention are used to measure the first yeast selected from saf-instant brands, and the measurement results are shown in the table 1 and 4.

FIG. 7B shows a schematic diagram of a microscopic image of the light-trapping microorganism system and its method for measuring the second yeast in a preferred embodiment of the present invention. Please refer to Figure 7B, the light-trapping microorganism system and method thereof and the light-trapping microorganism identification method of the preferred embodiment of the present invention are used to measure the second yeast selected from the brown brand. The measurement results are shown in Table 2 and 4.

FIG. 7C shows a schematic diagram of a microscopic image of the light-trapping microorganism system and its method for measuring the third yeast in a preferred embodiment of the present invention. Please refer to Figure 7C, the light trapping microorganism system and method and the light trapping microorganism identification method of the preferred embodiment of the present invention are used to measure the third yeast selected from the fermipan brand. The measurement results are shown in Table 3 and 4.

As shown in Table 4, different types of yeast have different escape speed and capture efficiency characteristics. Therefore, when the first, second and third yeasts are mixed, the optical tweezers system can be used to capture the mixed yeast, and after obtaining its escape speed and capture efficiency, the captured yeast can be known [brand] species.

Figure 8 shows a schematic diagram of the light-trapping microorganism identification method of the preferred embodiment of the present invention used to measure the first, second and third yeasts to obtain their escape speed. Please refer to Figure 8 and Table 4, the escape speed of the first yeast is 25.58 to 29.95 μm/s, the escape speed of the second yeast is 12.22 to 14.40 μm/s, and the escape speed of the third yeast is 8.49 To 10.4μm/s.

Fig. 9 shows a schematic diagram of the light trapping microorganism identification method used in measuring the first, second and third yeasts to obtain the trapping efficiency of the preferred embodiment of the present invention. Please refer to Figure 9 and Table 4, the capture efficiency of the first yeast is 3.17 to 3.46%, the capture efficiency of the second yeast is 2.26 to 2.80%, and the capture efficiency of the third yeast is 1.43 to 1.82%.

The above-mentioned experimental data are preliminary experimental results obtained under specific conditions, which are only used to easily understand or refer to the technical content of the present invention, and other related experiments are needed. The experimental data and results are not intended to limit the scope of rights of the present invention.

The foregoing preferred embodiments only illustrate the present invention and its technical features. The technology of this embodiment can still be implemented with various substantially equivalent modifications and/or alternatives; therefore, the scope of rights of the present invention is subject to patent application. The scope defined by the scope shall prevail. The copyright in this case is restricted to the use of patent applications in the Republic of China.

1‧‧‧Computer system

2‧‧‧Image capture system

21‧‧‧Microscope device

22‧‧‧Filter

3‧‧‧Laser light source device

31‧‧‧Laser light source

32‧‧‧Fiber Optic Coupler

33‧‧‧Optical fiber

34‧‧‧Fiber Microlens

5‧‧‧Working platform

51‧‧‧Pan stage control device

52‧‧‧The first electronic control platform

53‧‧‧ Stage

54‧‧‧Light source of stage

55‧‧‧Second Electronic Control Platform

Claims (10)

A light capturing microorganism system, comprising: a microorganism container for accommodating a microorganism; at least one microscope device corresponding to the microorganism container so that the microscope device can be used to display the microorganism of the microorganism container; and at least one Optical fiber optical tweezers, which uses the optical fiber optical tweezers to capture at least one microorganism with a predetermined light beam in the microorganism container, so as to form a captured microorganism on the optical fiber optical tweezers; wherein the optical fiber optical tweezers and The captured microorganisms move at at least a predetermined speed, and the optical fiber optical tweezers and the captured microorganisms are displayed through the microscope device, and the optical fiber optical tweezers and the captured microorganisms are moved from a first position to a second position. Two positions. According to the light capturing microorganism system described in claim 1, wherein the microscope device is connected to at least one image capturing system, and the microscope device corresponds to the microorganism, so that the image capturing system captures one from the microscope device Microorganism image. According to the light-trapping microorganism system described in item 1 of the scope of patent application, which further includes a working platform and a translation stage control device, the microorganism container is arranged on the working platform, and the translation stage control device is used to control the Work platform. According to the light-trapping microorganism system described in item 1 of the scope of patent application, it also includes a computer system, and the computer system is connected to the microscope device, or the computer system is connected to a laser light source device, or the computer system is connected to A working platform, a translation stage control device or an image capture system. According to the light-trapping microorganism system described in item 1 of the scope of patent application, the microorganism is selected from a yeast, a probiotic or a pathogenic bacteria. A method for capturing microorganisms by light, comprising: providing a microorganism container, and the microorganism container is used for accommodating a microorganism; providing at least one microscope device, and the microscope device corresponds to the microorganism Object container to display the microorganisms of the microorganism container by the microscope device; provide at least one optical fiber optical tweezers, and use the optical fiber optical tweezers to capture at least one microorganism in the microorganism container with a predetermined light beam, so as to A captured microorganism is formed on the optical tweezers; and the optical fiber optical tweezers and the captured microorganisms are controlled to move at at least a predetermined speed, and the optical fiber optical tweezers and the captured microorganisms are displayed through the microscope device, and The fiber optic tweezers and the captured microorganism move from a first position to a second position. According to the method of light capturing microorganisms described in the scope of patent application, the microscope device is connected to at least one image capturing system, and the microscope device corresponds to the microorganism, so that the image capturing system captures one from the microscope device Microorganism image. According to the method of light capturing microorganisms described in item 6 of the scope of patent application, it further includes a working platform and a translation stage control device, and the microorganism container is arranged on the working platform, and the translation stage control device is used to control the Work platform. According to the method of light capturing microorganisms described in item 6 of the scope of patent application, which further includes a computer system, and the computer system is connected to the microscope device, or the computer system is connected to a laser light source device, or the computer system is connected to A working platform, a translation stage control device or an image capture system. According to the method for capturing microorganisms by light according to item 6 of the scope of patent application, the microorganism is selected from a yeast, a probiotic or a pathogenic bacteria.

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