CN114384050A

CN114384050A - Biological particle positioning sensing method and system

Info

Publication number: CN114384050A
Application number: CN202011106916.7A
Authority: CN
Inventors: 邱子耕; 朱育娴; 欧阳自坤; 杨正仿
Original assignee: Individual
Current assignee: Li Ke Ming Bai Biotherapy Co ltd
Priority date: 2020-10-16
Filing date: 2020-10-16
Publication date: 2022-04-22

Abstract

A biological particle localization sensing method comprising: providing a carrier disc with a plurality of detection areas defined; adding a biological particle sample into the carrying disc; the sample of biological particles comprises a first biological particle having a biomarker and interacts with a corresponding tag; providing excitation energy to cause labels on the first biological particles to emit radiation energy; moving the first sensor to the detection areas respectively, defining the detection area receiving the radioactive energy as an active detection area after receiving the radioactive energy, and transmitting the position information of the active detection area to the processing module; according to the position information, the second sensor is moved to the detection area, the accurate position of the first biological particles in the activity detection area is detected, and the accurate position is transmitted to the processing module. A biological particle localization sensing system is also provided. By the method and the system, the specific biological particles can be rapidly detected, so that the detection efficiency is improved.

Description

Biological particle positioning sensing method and system

Technical Field

The invention relates to a biological particle positioning sensing method and a biological particle positioning sensing system; in particular to a method and a system for positioning and sensing biological particles, which can improve the detection efficiency of biological particles.

Background

With the development of science and technology, the detection technology of biomedicine is also improved, and the detection accuracy can be improved by utilizing an immune mode for specific biological particles. In the past, biological particles have been detected with high precision mainly by means of immunofluorescence coloration; in detail, immunofluorescence coloration is directed to a specific surface antigen of a biological particle, and is specifically recognized by a fluorescent material with an antibody, so that the target biological particle emits fluorescence under the irradiation of light with a specific wavelength, and is detected by a sensor.

However, for samples with large sample quantity or weak fluorescence, the traditional detection method for biological particles cannot effectively complete detection; for example, in the conventional biological particle detection device, a charge coupled device or a photosensitive device is used as a sensor, an image under each field of view is captured by the sensor, the images are pieced into a complete image through software operation, and then a specific fluorescence signal is identified through intelligent software, so that the detection limit value of the sensor is high, and therefore, a sample with weak fluorescence cannot be accurately detected, and has high-proportion misalignment; in addition, the sensor requires a long time for sensing, so that the detection cannot be completed quickly for a large number of samples, and the detection efficiency is low.

Accordingly, there is a need for a novel method and system for positioning and sensing biological particles to solve the long-standing technical problems of the conventional detection methods and systems.

Disclosure of Invention

In view of the above, the present invention provides a method and system for positioning and sensing biological particles, which uses a first sensor to quickly and sharply screen out a detection area having specific biological particles, records the position information of the detection area, and then a second sensor moves to the detection area having specific biological particles according to the position information, performs a high-precision detection procedure on the specific biological particles in the detection area, and records the accurate position and related information of the specific biological particles in an active detection area.

In order to achieve the above object, the present invention provides a method for positioning and sensing biological particles, comprising at least the following steps:

providing a carrying disc, and defining a plurality of detection areas on the carrying disc;

adding a sample of biological particles to the boat; the biological particle sample comprises a first biological particle, wherein the first biological particle has at least one biomarker and the at least one biomarker interacts with a corresponding at least one tag;

providing an excitation energy to the carrier by an excitation device, so that the at least one label on the first biological particle emits a radiation energy;

relatively moving a first sensor to the plurality of detection areas respectively; when the first sensor receives the radiation energy, defining the detection area receiving the radiation energy as an active detection area, and transmitting position information of the active detection area to a processing module; and

according to the position information, a second sensor is corresponding to the active detection area, an accurate position of the first biological particles in the active detection area is detected, and the accurate position is transmitted to the processing module.

Another objective of the present invention is to provide a positioning and sensing system for biological particles, which comprises a carrier, an excitation device, a first sensor and a second sensor; wherein the carrier plate is defined with a plurality of detection areas, and a biological particle sample is arranged in the carrier plate; the biological particle comprises a first biological particle, wherein the first biological particle has at least one biomarker; the at least one biomarker interacts with a corresponding at least one tag; the excitation device is operably moved to be close to the carrier and provides an excitation energy toward the carrier, so that the at least one label on the first biological particle emits a radiation energy; the first sensor can be controlled to move relative to the carrying disc and respectively move to the plurality of detection areas; when the first sensor receives the radiation energy, defining the detection area receiving the radiation energy as an active detection area, and transmitting position information of the active detection area to a processing module; the second sensor moves to the active detection area relative to the carrier disc according to the position information, detects an accurate position of the first biological particles in the active detection area, and transmits the accurate position to the processing module.

The present invention has the effects that the first sensor is used to quickly and sharply screen out the detection area with the specific biological particles and record the position information of the detection area, then the second sensor moves to the detection area with the specific biological particles according to the position information, the high-precision detection step is carried out on the specific biological particles in the detection area, and the accurate position and the relevant information of the specific biological particles in the activity detection area are recorded. Therefore, the biological particle positioning and sensing method and the biological particle positioning and sensing system can improve the detection efficiency of the biological particles, have high precision and solve the problems that the traditional biological detection device and the traditional biological detection method cannot effectively detect samples with large quantity or weak fluorescence.

Drawings

FIG. 1 is a flowchart of a method for positioning and sensing biological particles according to a preferred embodiment of the present invention.

Fig. 2 is a complementary flow diagram to fig. 1.

FIG. 3 is a top view of a biological particle localization sensing system according to a preferred embodiment of the present invention.

Figure 4 is a cross-sectional view taken along line 4-4 of figure 3.

FIG. 5 is another cross-sectional view of a biological particle localization sensing system according to a preferred embodiment of the present invention.

FIG. 6 is a cross-sectional view of a biological particle localization sensing system according to another preferred embodiment of the present invention.

FIG. 7 is a cross-sectional view of a biological particle localization sensing system according to another preferred embodiment of the present invention.

Detailed Description

In order to more clearly illustrate the present invention, preferred embodiments are described in detail below with reference to the accompanying drawings. Referring to fig. 1 to 6, fig. 1 is a flowchart illustrating a method for positioning and sensing biological particles according to a preferred embodiment of the present invention; FIG. 2 is a complementary flow diagram to FIG. 1; FIG. 3 is a top view of a biological particle localization sensing system according to a preferred embodiment of the present invention; FIG. 4 is a cross-sectional view taken along line 4-4 of FIG. 3; FIG. 5 is another cross-sectional view of a biological particle localization sensing system according to a preferred embodiment of the present invention; FIG. 6 is a cross-sectional view of a biological particle localization sensing system according to another preferred embodiment of the present invention.

Referring to fig. 4 and 5, a biological particle positioning and sensing system includes a carrier 10, an excitation device 20, a first sensor 30 and a second sensor 40. The carrier disc 10 is defined with a plurality of detection areas 12, and a biological particle sample BP1, BP2 and BP3 are arranged in the carrier disc 10; the biological particles BP1, BP2, BP3 comprise a first biological particle BP1, wherein the first biological particle BP1 has at least one biomarker (not shown); the at least one biomarker interacts with a corresponding at least one tag (not shown).

The excitation device 20 is operably moved to be close to the carrier 10 and provides an excitation energy towards the carrier 10, so that the at least one label on the first biological particle BP1 emits a radiation energy. The tag composition attached to the first biological particle BP1 absorbs the excitation energy and emits the radiation energy. In an embodiment of the present invention, the biological particles BP1, BP2, BP3 comprise a second biological particle BP2, BP3, and the second biological particle BP2, BP3 does not interact with the at least one label, so the second biological particle BP2, BP3 does not generate the radioactive energy. In another embodiment of the present invention, the immune matching relationship between the at least one biomarker of the first biological particle and the corresponding at least one label and the mechanism of action for exciting the fluorescent molecule on the at least one label to emit fluorescence may be applied to Fluorescence Resonance Energy Transfer (FRET) and Bioluminescence Resonance Energy Transfer (BRET).

The first sensor 30 is operable to move relative to the boat 10 and to the plurality of detection areas 12; when the first sensor 30 moves to one of the detection regions 12 and the first sensor 30 receives the radiation energy, the first sensor 30 defines the detection region 12 receiving the radiation energy as an active detection region 121 and transmits a position information of the active detection region 121 to a processing module P. The second sensor 40 moves to the active detection area 121 according to the position information, detects an accurate position 122 of the first biological particle BP1 in the active detection area 121, and transmits the accurate position 122 to the processing module P. In another embodiment of the present invention, the tray is operable to move relative to the first sensor, and the plurality of detection regions are moved to correspond to the first sensor respectively.

Referring to fig. 1, the method for sensing the positioning of biological particles includes at least the following steps:

step S1: providing a carrying disc 10, and defining a plurality of detection areas 12 on the carrying disc 10;

step S3: adding a biological particle sample BP1, BP2, BP3 to the carrier disc 10; the biological particle samples BP1, BP2, BP3 comprise a first biological particle BP1, wherein the first biological particle BP1 has at least one biomarker (not shown) and the at least one biomarker interacts with corresponding at least one label (not shown);

step S5: providing an excitation energy to the boat 10 by an excitation device 20, so that the at least one label on the first biological particle BP1 emits a radiation energy;

step S7: relatively moving a first sensor 30 to the plurality of detection areas 12; when the first sensor 30 moves to one of the detection regions 12 and the first sensor 30 receives the radiation energy, the first sensor 30 defines the detection region 12 receiving the radiation energy as an active detection region 121 and transmits a position information of the active detection region 121 to a processing module P; and

step S9: according to the position information, a second sensor 40 is assigned to the active detection region 121, detects an accurate position 122 of the first biological particle BP1 in the active detection region 121, and transmits the accurate position 122 to the processing module P.

According to the embodiment of the present invention, in step S3, the biological particles BP1, BP2, BP3 include a second biological particle BP2, BP3, and the second biological particle BP2, BP3 does not interact with the at least one label, so that in step S5, the second biological particle BP2, BP3 does not generate the radioactive energy, as shown in fig. 5. The biological particles BP1, BP2, BP3 include, but are not limited to, cells, bacteria, fungi, algae, protozoa, worms, viruses, protein vectors, nucleic acid vectors, or combinations thereof.

According to an embodiment of the invention, the at least one biomarker comprises at least one nucleic acid, protein or polysaccharide molecule located on or within a biological particle, and the at least one tag comprises a protein, nucleic acid, polysaccharide molecule or specific compound. In another embodiment of the invention, the at least one biomarker comprises at least one surface antigen located on the biological particle, and the at least one tag comprises at least one antibody or chemical molecular stain to specifically recognize the at least one surface antigen or an intercellular, intracellular protein or nucleic acid; in addition, the at least one label includes a luminescent structure, a fluorescent structure, a phosphorescent structure, a physically distinguishable structure, a chemically distinguishable structure, or a combination thereof. According to embodiments of the invention, the at least one biomarker includes EpCAM, CD45, CD71, GPA, nucleic acids, or combinations thereof, but is not limited thereto, and any surface antigen that may be present on the cell surface may be suitable for use in embodiments of the invention. According to an embodiment of the present invention, the fluorescent structure comprises a fluorescent protein, a Quantum Dot particle (Quantum Dot), or a combination thereof. According to an embodiment of the invention, when the at least one biomarker is EpCAM, the excitation wavelength is in the range of 450nm to 500nm and the emission wavelength is in the range of 520nm to 555 nm; when the at least one biomarker is a nucleic acid, the excitation wavelength is in the range of 380nm to 420nm and the emission wavelength is in the range of 540nm to 560 nm; when the at least one biomarker is CD45, the excitation wavelength ranges from 600nm to 650nm and the emission wavelength ranges from 660nm to 720 nm. According to another embodiment of the present invention, compared to general fluorescent molecules, the quantum dot particles have a wider excitation wavelength range (more than 10 nm) and generate a narrower emission wavelength range, and the emission spectrum is more symmetrical, so as to generate a significant difference from the quantum dot particles in other emission wavelength ranges; in addition, the quantum dot particles can regulate the emission wavelength range through the particle size, so that the quantum dot particles with different particle sizes can be excited by using the excitation light sources with the same excitation wavelength range and emit the emitted light with different emission wavelength ranges, and the purpose of labeling and identifying various biomarkers can be achieved by using the same quantum dot material and the same excitation wavelength. The quantum dot particles also have the advantages of high fluorescence intensity, long fluorescence lifetime, good stability, good biocompatibility and the like, so in another embodiment of the invention, the quantum dot particles can be used for marking and identifying the biomarkers on or in the biological particles.

According to the embodiment of the present invention, the excitation device 20 includes a cold light source, a laser light source, an ultraviolet light source, a visible light source, an infrared light source, an ultrasonic generator, an electromagnetic wave generator, a microwave generator, or a combination thereof. In step S5, the excitation energy provided by the excitation device 20 toward the carrier 10 includes excitation light, excitation sound waves, excitation electromagnetic waves, or a combination thereof, and the radiation energy emitted by the at least one tag on the first biological particle BP1 includes optical signals, electrical signals, magnetic signals, acoustic signals, or a combination thereof.

In an embodiment of the present invention, the first sensor 30 includes a photomultiplier tube (PMT), a Charge Coupled Device (CCD), a photo resistor, an ultrasonic sensor, an induction coil, or a combination thereof; the first sensor 30 is preferably a photomultiplier tube (PMT) capable of detecting a weak light source, and in one embodiment of the present invention, the detection limit of the PMT may be less than or equal to 5V, such as 1V, 1.5V, 2V, 2.5V, 3V, 3.5V, 4V, 4.5V, or 5V; the second sensor 40 is preferably, but not limited to, a Charge Coupled Device (CCD), a Complementary Metal-Oxide Semiconductor (CMOS) or a combination thereof, which can provide high image resolution. In an embodiment of the invention, when the radiation energy is an optical signal, the optical signal has a first optical path to the first sensor and a second optical path to the second sensor, and the first optical path and the second optical path are the same or different. If the first optical path and the second optical path are different, the first optical path and the second optical path are independent optical paths respectively.

Referring to fig. 2 and 3, in step S7, relatively moving the first sensor 30 to the inspection regions 12 includes moving the first sensor 30 relative to the carrier tray along a moving path R (i.e., step S71), wherein the moving path R passes through the positions corresponding to the inspection regions 12 in a predetermined sequence (i.e., step S73). In the embodiment of the present invention, the moving path R may be a linear progressive scan (as shown in fig. 3) or a circular scan with the center of the carrier 10 as the center, or any scanning manner that can pass through the plurality of detection areas. In step S7, moving the first sensor 30 to the detection regions 12 respectively means moving the first sensor and the carrier plate relatively, i.e. fixing the carrier plate to move the first sensor relative to the carrier plate in an operable manner, or fixing the first sensor to move the carrier plate relative to the first sensor in an operable manner, to move the detection regions respectively to correspond to the first sensor.

In another embodiment of the present invention, the excitation device 20, the first sensor 30 and the second sensor 40 are disposed on the same robot arm 50, and when the robot arm 50 moves, the excitation device 20, the first sensor 30 and the second sensor 40 move synchronously, as shown in fig. 4 and 5; in other words, steps S5, S7, and S9 of the method for sensing the location of biological particles can be performed simultaneously, or step S5 can be performed in two stages in combination with step S7 and step S5 in combination with step S9. In fig. 4 and 5, the robot arm 50 can be controlled to move in the horizontal direction D1 and the vertical direction D2. In step S9, the tray is also operable to move relative to the second sensor, and the active detection regions are moved to correspond to the second sensor respectively.

In another embodiment of the present invention, the excitation device 20 and the first sensor 30 are disposed on a robot arm 50a, another excitation device 20 and the second sensor 40 are disposed on a robot arm 50b, and the excitation devices 20, the first sensor 30, the second sensor 40 and the

robot arms

50a, 50b are in signal connection with the processing module P, and the excitation devices 20, the first sensor 30, the second sensor 40 and the

robot arms

50a, 50b are controlled by the processing module P to move and displace, as shown in fig. 6; in other words, in the present embodiment, the method for sensing the location of biological particles can be performed by combining step S5 with step S7 and combining step S5 with step S9 in two stages. In fig. 6, the

robot arms

50a, 50b are controllably movable in a horizontal direction D1 and a vertical direction D2.

In fig. 4, the boat 10 has a transparent bottom 14, and the excitation device 20, the first sensor 30 and the second sensor 40 move under the transparent bottom 14 of the boat 10. The tray 10 is disposed in an accommodating groove 602 of a supporting platform 60, the accommodating groove 602 has an opening, and the transparent bottom 14 of the tray 10 corresponds to the opening. In addition, in another embodiment of the present invention, the excitation device, the first sensor and the second sensor move above the boat.

In the embodiment of the present invention, a light path formed between the carrying stage 60 and the first sensor 30 and a light path formed between the carrying stage 60 and the second sensor 40 are respectively provided with a filter set, so as to improve the sensing sensitivity and accuracy of the first sensor 30 and the second sensor 40.

For example, in fig. 6, the carrier 60 includes a filter 62 covering the opening, and when the excitation device 20, the first sensor 30 and the second sensor 40 move under the carrier 10, the excitation energy and the radiation energy transmit the filter 62, so that the first sensor 30 and the second sensor 40 receive the radiation energy. It should be noted that, since the wavelength emitted by the laser is different from the wavelength released after the fluorescent molecules on the at least one label are excited by the laser, the filter set 62 disposed at the opening at least includes a combination of a lens, a filter and a reflector, so that the excitation light (i.e. the laser light) and the emission light (i.e. the fluorescence) can pass through different light paths of the filter set 62, and the sensing sensitivity and accuracy of the first sensor 30 and the second sensor 40 are improved by the filter set 62.

In another embodiment of the present invention, in fig. 7, the first sensor 30 includes a filter 32 covering the light incident surface of the first sensor 30, and the second sensor 40 includes a filter 42 covering the light incident surface of the second sensor 40. When the excitation device 20, the first sensor 30 and the second sensor 40 are actuated under the boat 10, the excitation energy and the radiation energy are transmitted through the filter sets 32, 42, so that the first sensor 30 and the second sensor 40 receive the radiation energy. It should be noted that, since the wavelength emitted by the laser is different from the wavelength released after the fluorescent molecules on the at least one label are excited by the laser, the filter set 32, 42 disposed at the opening at least includes the combination of the lens, the filter and the reflector, so that the excitation light (i.e. the laser light) and the emission light (i.e. the fluorescence) can pass through different light paths of the filter set 32, 42, and the sensing sensitivity and accuracy of the first sensor 30 and the second sensor 40 are improved by the filter set 32, 42.

In an embodiment of the present invention, the second sensor 40 is preferably a high-speed charge-coupled device (high-speed CCD) capable of providing high image resolution, and the second sensor 40 can be used to confirm that the first biological particles BP1 are present at the accurate position 122 in the active detection region 121 on the carrier tray 10 and feed back information of the accurate position 122 of the active detection region 121 to the processing module P. The first biological particles BP1 may be one or more, and the second sensor 40 may further capture a clear image of the first biological particles BP 1.

In one embodiment of the present invention, one or more surface antigens on the first biological particle BP1 are bound by one or more antibodies, one or more fluorescent molecules on the one or more antibodies can be excited by a specific radiation source and release a specific fluorescent signal, a photomultiplier tube (PMT) detects the fluorescent signal to primarily confirm the active detection region 121 where the first biological particle BP1 exists, then a charge-coupled device (CCD) captures different fluorescent signals to confirm the accurate position 122 of the first biological particle BP1, and the accurate position 122 is used to sort the first biological particle BP1 and other biological particles BP2 and BP 3.

In one embodiment of the invention, the first biological particle BP1 may be a Circulating Tumor Cell (CTC) on which surface antigens may be recognized by at least one antibody with a fluorescent molecule. For example, an EpCAM surface antigen on a Circulating Tumor Cell (CTC) can bind to an EpCAM antibody to generate a first fluorescent signal, and a Hoechst antibody can bind to a nucleic acid within the Circulating Tumor Cell (CTC) to generate a second fluorescent signal; however, since the Circulating Tumor Cells (CTCs) do not have CD45 antigen thereon and cannot bind to the CD45 antibody, a third fluorescence signal is not generated, so that if the first fluorescence signal and the second fluorescence signal are detected in a detection region but the third fluorescence signal is not detected, the first biomicron BP1, i.e., the Circulating Tumor Cells (CTCs), is determined to be present in the detection region, and the Photomultiplier (PMT) returns the position of the detection region 12 where the Circulating Tumor Cells (CTCs) are present to the processing module P.

In another embodiment of the present invention, if the detection region of the second biological particle BP2 does not allow the photomultiplier tube (PMT) to detect the first fluorescence signal or the second fluorescence signal, or allows the photomultiplier tube (PMT) to detect the third fluorescence signal, it is determined that the second biological particle BP2 is not a Circulating Tumor Cell (CTC), i.e., the first biological particle BP1 is not present in the detection region, and the photomultiplier tube (PMT) skips over the detection region of the second biological particle BP 2. Alternatively, if the detection region of the third biological particle BP3 allows the photomultiplier tube (PMT) to detect only one of the first and second fluorescent signals but not the other of the first and second fluorescent signals, the third biological particle BP3 is determined to be not a Circulating Tumor Cell (CTC), and the photomultiplier tube (PMT) will skip the detection region of the third biological particle BP 3. Therefore, in the embodiment of the present invention, fluorescence signals of the detection regions where the biological particles BP1, BP2, BP3 are located can be rapidly scanned and detected by a photomultiplier tube (PMT), and whether the biological particles BP1, BP2, BP3 are present in the detection regions can be determined in real time, so as to immediately perform recording and positioning on the active detection region 121 where the first biological particle BP1 is present.

Then, after the photomultiplier tube (PMT) returns the position of the active detection area 121 where the Circulating Tumor Cells (CTCs) are present to the processing module P and excludes the detection area where no Circulating Tumor Cells (CTCs) are present, the Charge Coupled Device (CCD) moves to the active detection area 121. If a Charged Coupled Device (CCD) detects a biological particle with a first and a second fluorescence signal but no third fluorescence signal in the field of view of the active detection area 121, it is determined that the detected first biological particle BP1 is a Circulating Tumor Cell (CTC), and the exact location 122 of the Circulating Tumor Cell (CTC) in the active detection area 121 is returned to the processing module P, and the first biological particle BP1 is resolved from the exact location 122 fed back by the Charged Coupled Device (CCD).

According to the embodiment of the invention, the first sensor is used for quickly and sharply screening out the detection area with the specific biological particles, the position information of the detection area is recorded, then the second sensor moves to the detection area with the specific biological particles according to the position information, the high-precision detection step is carried out on the specific biological particles in the detection area, and the accurate position and the related information of the specific biological particles in the activity detection area are recorded. Therefore, the biological particle positioning and sensing method and the biological particle positioning and sensing system can improve the detection efficiency of the biological particles, have high precision and solve the problems that the traditional biological detection device and the traditional biological detection method cannot effectively detect samples with large quantity or weak fluorescence.

The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications to the present invention as described and claimed should be included in the scope of the present invention.

Description of the reference numerals

[ invention ]

10: carrying disc

12: detection area

121: active detection area

122: accurate position

14: transparent bottom

20: excitation device

30: first sensor

40: second sensor

50. 50a, 50b, 50 c: mechanical arm

60: bearing platform

602: containing groove

62. 32, 42: filter lens group

4-4: section line

BP1, BP2, BP 3: biological particles

D1: in the horizontal direction

D2: in the vertical direction

P: processing module

R: moving path

S1, S3, S5, S7, S9, S71, S73: step (ii) of

Claims

1. A biological particle localization sensing method, comprising:

according to the position information, a second sensor is corresponding to the detection area, an accurate position of the first biological particles in the detection area is detected, and the accurate position is transmitted to the processing module.

2. The method for sensing localization of biological particles as recited in claim 1, wherein the biological particle includes a second biological particle that does not interact with the at least one tag.

3. The method as claimed in claim 1, wherein moving the first sensor to the detection regions respectively causes the first sensor to travel relative to the carrier plate along a moving path that passes through positions corresponding to the detection regions in a predetermined sequence.

4. The biological particle localization sensing method of claim 1, wherein the biological particles comprise cells, bacteria, fungi, algae, protozoa, worms, viruses, protein vectors, nucleic acid vectors, or a combination thereof.

5. The method for sensing localization of biological particles as claimed in claim 1, wherein the at least one biomarker includes at least one surface antigen on the first biological particle and the at least one tag includes at least one antibody to specifically recognize the at least one surface antigen.

6. The method of claim 1, wherein the at least one tag comprises a luminescent structure, a fluorescent structure, a phosphorescent structure, a physically distinguishable structure, a chemically distinguishable structure, or combinations thereof.

7. The biological particle localization sensing method of claim 6, wherein the fluorescent structure comprises a fluorescent protein, a quantum dot particle, or a combination thereof.

8. The method of claim 6, wherein the excitation device comprises a cold light source, a laser light source, an ultraviolet light source, a visible light source, an infrared light source, an ultrasonic generator, an electromagnetic generator, a microwave generator, or combinations thereof.

9. The method of claim 8, wherein the radiated energy comprises an optical signal, an electrical signal, a magnetic signal, an acoustic signal, or a combination thereof.

10. The biological particle localization sensing method of claim 9, wherein the first sensor comprises a photomultiplier tube, a charge coupled device, a photoresistor, an ultrasonic sensor, an induction coil, or a combination thereof.

11. The method of claim 9, wherein the second sensor comprises a charge coupled device, a complementary metal oxide semiconductor device, or a combination thereof.

12. The method as claimed in claim 9, wherein when the emission energy is an optical signal, the optical signal has a first optical path to the first sensor and a second optical path to the second sensor, and the first and second optical paths are the same or different.

13. The method as claimed in claim 1, wherein the excitation device, the first sensor and the second sensor are disposed on a same robot arm, and the excitation device, the first sensor and the second sensor move synchronously when the robot arm moves.

14. The method as claimed in claim 1, wherein the excitation device and the first sensor are disposed on a robot arm, another excitation device and the second sensor are disposed on another robot arm, and the excitation devices, the first sensor and the second sensor, and the robot arms are connected to the processing module via signals, and the excitation devices, the first sensor and the second sensor, and the robot arms are controlled by the processing module to move and displace.

15. The method of claim 1, wherein the boat has a transparent bottom, and the excitation device, the first sensor and the second sensor move under the transparent bottom of the boat.

16. The method as claimed in claim 15, wherein the tray is disposed in a receiving cavity of a carrier, the receiving cavity has an opening, and the transparent bottom of the tray corresponds to the opening; the bearing table comprises a filter group covering the opening, and when the excitation device, the first sensor and the second sensor act below the bearing disc, the excitation energy and the radiation energy transmit the filter group, so that the first sensor and the second sensor receive the radiation energy.

17. A biological particle localization sensing system, comprising:

a carrying disc, which is defined with a plurality of detection areas, and a biological particle sample is arranged in the carrying disc; the biological particle comprises a first biological particle, wherein the first biological particle has at least one biomarker; the at least one biomarker interacts with a corresponding at least one tag;

an excitation device operably moved to be close to the carrier and providing an excitation energy toward the carrier to cause the at least one label on the first biological particle to emit a radiation energy;

a first sensor, which can be controlled to move relative to the carrying disc and respectively move to the plurality of detection areas; when the first sensor receives the radiation energy, defining the detection area receiving the radiation energy as an active detection area, and transmitting position information of the active detection area to a processing module; and

a second sensor, moving to the active detection area relative to the carrier according to the position information, detecting an accurate position of the first biological particles in the detection area, and transmitting the accurate position to the processing module.

18. The biological particle localization sensing system of claim 17, wherein the biological particle comprises a second biological particle that does not interact with the at least one tag.

19. The system as claimed in claim 17, wherein moving the first sensor to the detection zones respectively causes the first sensor to travel relative to the carrier plate along a path of travel that passes through positions corresponding to the detection zones in a predetermined sequence.

20. The biological particle localization sensing system of claim 17, wherein the biological particles comprise cells, bacteria, fungi, algae, protozoa, worms, viruses, protein carriers, nucleic acid carriers, or a combination thereof.

21. The biological particle localization sensing system of claim 17, wherein the at least one biomarker comprises at least one surface antigen on the first biological particle and the at least one label comprises at least one antibody to specifically recognize the at least one surface antigen.

22. The biological particle localization sensing system of claim 17, wherein the at least one tag comprises a luminescent structure, a fluorescent structure, a phosphorescent structure, a physically distinguishable structure, a chemically distinguishable structure, or a combination thereof.

23. The biological particle localization sensing system of claim 22, wherein the fluorescent structure comprises a fluorescent protein, a quantum dot particle, or a combination thereof.

24. The system of claim 22, wherein the excitation device comprises a cold light source, a laser light source, an ultraviolet light source, a visible light source, an infrared light source, an ultrasonic generator, an electromagnetic generator, a microwave generator, or combinations thereof.

25. The biological particle localization sensing system of claim 24, wherein the radiated energy comprises an optical signal, an electrical signal, a magnetic signal, an acoustic signal, or a combination thereof.

26. The biological particle localization sensing system of claim 25, wherein the first sensor comprises a photomultiplier tube, a charge coupled device, a photoresistor, an ultrasonic sensor, an induction coil, or a combination thereof.

27. The biological particle localization sensing system of claim 25, wherein the second sensor comprises a charge coupled device, a complementary metal oxide semiconductor device, or a combination thereof.

28. The biological particle localization sensing system of claim 25, wherein when the radiant energy is an optical signal, the optical signal has a first optical path to the first sensor and a second optical path to the second sensor, the first and second optical paths being the same or different.

29. The system as claimed in claim 17, wherein the excitation device, the first sensor and the second sensor are disposed on a same robot arm, and the excitation device, the first sensor and the second sensor move synchronously when the robot arm moves.

30. The system as claimed in claim 17, wherein the excitation device and the first sensor are disposed on a robot arm, another excitation device and the second sensor are disposed on another robot arm, and the excitation devices, the first sensor and the second sensor, and the robot arms are in signal connection with the processing module, and the excitation devices, the first sensor and the second sensor, and the robot arms are controlled by the processing module to move and displace.

31. The biological particle localization sensing system of claim 17, wherein the boat has a transparent bottom, and the excitation device, the first sensor, and the second sensor move under the transparent bottom of the boat.

32. The system as claimed in claim 31, wherein the tray is disposed in a receiving cavity of a carrier, the receiving cavity has an opening, and the transparent bottom of the tray corresponds to the opening; the bearing table comprises a filter group covering the opening, and when the excitation device, the first sensor and the second sensor act below the bearing disc, the excitation energy and the radiation energy transmit the filter group, so that the first sensor and the second sensor receive the radiation energy.