Detection device for detecting biological particles and detection method of detection device

Abstract

A detection device for detecting biological particles includes an optical system, which includes an excitation light source, a filter spectroscope group, a photomultiplier tube and a charge-coupled device. The excitation light source irradiates the biological particles on a detection stage of the detection device, and a target biological particle among the multiple biological particles is excited to generate a radiation light. The radiation light is incident on the filter spectroscope group and is divided into a first detection light and a second detection light. After receiving the first detection light, the photomultiplier tube transmits a regional positioning signal to a processor of the detection device, and after receiving the second detection light, the charge-coupled device transmits an image signal to the processor. In this way, the processor can obtain the precise position of the target biological particle according to the regional positioning signal and the image signal. In addition, a detection method of the above-mentioned detection device is also provided herein.

Description

Technical Field

[0001] The present invention relates to an inspection and measurement system; in particular, it relates to a detection device for detecting biological particles and a detection method of the detection device. Background Art

[0002] Nowadays, optical sensors are widely used in measurement systems in various fields. Optical sensors can perform a wide range of detection and imaging characteristics, making them widely used in the field of biomedical imaging. In the biomedical field, optical sensor fluorescence imaging is often used to facilitate image monitoring or scanning detection.

[0003] The fluorescence imaging technology of the known scanning detection device must have a lens group, a scanning sensor and a light source. The light source emits a high-penetration light, such as near-infrared light or laser, to illuminate the biological particles on the device carrier, so that at least part of the biological particles will emit a radiation light after being illuminated, and the radiation light emitted by these biological particles can be emitted into the optical sensor after filtering the light wave through the lens group, so that the optical sensor can capture the image of the biological particles and locate the position of each biological particle, so as to analyze and observe the activity of the target biological particles, and even further select them.

[0004] However, in order to obtain better image resolution, current scanning detection devices usually choose to add a lens group, scanning sensor and light source combination near the device stage, but the above-mentioned assembled scanning detection device is prone to mechanical motion interference between one scanning sensor and another scanning sensor due to the poor integration of their respective moving devices; furthermore, the structure of the above-mentioned assembled scanning detection device is too complicated and numerous, resulting in a large overall size and weight. Therefore, the above-mentioned problem really needs to be solved. Summary of the invention

[0005] In view of this, an object of the present invention is to provide a detection device for detecting biological particles and a detection method for the detection device, which has the characteristics of reduced overall device volume and weight and precise movement path during detection, while efficiently achieving the positioning and identification of biological particles.

[0006] In order to achieve the above-mentioned purpose, the detection device for detecting biological particles provided by the present invention comprises a detection stage for placing a plurality of biological particles and an optical system. The plurality of biological particles include at least one target biological particle. The optical system comprises an excitation light source, a filter spectroscope group, at least one photomultiplier tube (PMT) and at least one charge coupled device (CCD). The excitation light source is used to provide an excitation light to illuminate the plurality of biological particles, and the at least one target biological particle among the plurality of biological particles is illuminated by the excitation light and excited to generate a radiation light. The filter spectroscope group comprises a spectroscope, wherein the spectroscope is located on the optical path of the radiation light, and is used to divide the incident radiation light into a first detection light and a second detection light that account for different proportions of the radiation light, wherein the ratio of the first detection light to the radiation light is between 0.1 and 0.5, and the ratio of the second detection light to the radiation light is between 0.5 and 0.9. The at least one photomultiplier tube (PMT) is used to receive the first detection light and can move relative to the detection stage; when receiving the first detection light, the at least one photomultiplier tube generates a regional positioning signal, and the regional positioning signal includes the regional position of the at least one target biological particle on the detection stage. The at least one charge-coupled device (CCD) is used to receive the second detection light and can move relative to the detection stage; when receiving the second detection light, the at least one charge-coupled device generates an image signal, and the image signal includes the image position of the at least one target biological particle on the detection stage.

[0007] In order to achieve the above object, the present invention further provides a detection method for a detection device, which is used for the above detection device, and the detection device further includes a processor, and the processor is electrically connected to the optical system. The detection method includes the following steps:

[0008] Step A: the excitation light source emits the excitation light to illuminate the plurality of biological particles;

[0009] Step B: the at least one target biological particle among the plurality of biological particles absorbs the excitation light to generate the radiated light, and the radiated light is incident on the filter spectroscope group;

[0010] Step C: the filter spectroscope group separates the radiated light into the first detection light and the second detection light, and the first detection light and the second detection light are incident on the at least one photomultiplier tube and the at least one charge coupled device respectively;

[0011] Step D: the at least one photomultiplier tube receives the first detection light and generates a regional positioning signal;

[0012] Step E: the processor determines that at least one area position on the detection stage has at least one target biological particle according to the area positioning signal;

[0013] Step F: the at least one charge coupled device receives the second detection light and generates an image signal;

[0014] Step G: The processor obtains the precise position of the at least one target biological particle in the at least one regional position on the detection stage according to the image signal.

[0015] The effect of the present invention is that the detection device for detecting biological particles of the present invention only needs to use an excitation light source and a set of filter spectroscopes to simultaneously locate the precise position of the target biological particles with a photomultiplier tube and a charge-coupled device. In addition, the photomultiplier tube of the present invention first quickly scans and determines the regional position of the target biological particles. After the target biological particles are found, the charge-coupled device then locates the precise position of the target biological particles in detail, thereby achieving the effect of high-efficiency detection and high detection accuracy. The mechanical structure of the integrated photomultiplier tube and the charge-coupled device makes the action path during scanning detection more accurate, and at the same time, there is more room for use in the mechanism design. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a schematic diagram of a detection device for detecting biological particles according to a first preferred embodiment of the present invention.

[0017] Figure 2 This is another schematic diagram of the detection device for detecting biological particles according to the preferred embodiment of the present invention, which discloses the first light path.

[0018] Figure 3 This is another schematic diagram of the detection device for detecting biological particles according to the preferred embodiment of the present invention, revealing the second optical path.

[0019] Figure 4 It is a schematic diagram of another aspect of the detection device for detecting biological particles according to the above preferred embodiment of the present invention.

[0020] Figure 5 Schematic diagram of a detection device for detecting biological particles according to a second preferred embodiment of the present invention.

[0021] Figure 6 It is a schematic diagram of a detection device for detecting biological particles according to a third preferred embodiment of the present invention.

[0022] Figure 7 Schematic diagram of a detection device for detecting biological particles according to a fourth preferred embodiment of the present invention.

[0023] Figure 8 Schematic diagram of a detection device for detecting biological particles according to a fifth preferred embodiment of the present invention.

[0024] Fig. 9 Schematic diagram of a detection device for detecting biological particles according to a sixth preferred embodiment of the present invention.

[0025] Fig.10 Schematic diagram of a detection device for detecting biological particles according to the seventh preferred embodiment of the present invention.

[0026] Fig.11 Schematic diagram of a detection device for detecting biological particles according to an eighth preferred embodiment of the present invention.

[0027] Fig.12 It is a schematic diagram of a detection device for detecting biological particles according to the ninth preferred embodiment of the present invention.

[0028] Fig.13 Schematic diagram of a detection device for detecting biological particles according to the tenth preferred embodiment of the present invention.

[0029] Fig.14 It is a schematic diagram of a detection device for detecting biological particles according to the eleventh preferred embodiment of the present invention. DETAILED DESCRIPTION

[0030] In order to more clearly illustrate the present invention, preferred embodiments are described in detail below with reference to the accompanying drawings. Figures 1 to 3 As shown, the detection device 100 of the first preferred embodiment of the present invention includes a detection platform 10, an optical system (not shown) and a processor (not shown) electrically connected to the optical system, wherein the optical system includes an excitation light source 20, a filter spectrometer group 30 and a scanning sensor group 40.

[0031] The detection carrier 10 is used to place a plurality of biological particles (not shown), and the biological particles include but are not limited to cells, bacteria, fungi, viruses, exosomes, liposomes, or nucleic acid-encapsulated carriers and other biological particles that are generally understandable to those skilled in the art, and the biological particles include at least one target biological particle, and the target biological particles include but are not limited to nucleic acids, proteins, lipids, glycoproteins and other substances. Any substance of the at least one target biological particle can be combined with at least one label, and the at least one label can be combined with the surface protein or the nucleic acid in the nucleus of the at least one target biological particle to serve as a research and detection object. The at least one label can be but is not limited to fluorescent particles, cold light particles or quantum dots. In this embodiment, the type of target biological particles is described as one example, and the target biological particles are combined with a label that is a fluorescent particle.

[0032] The excitation light source 20 includes but is not limited to laser, mercury lamp and LED lamp, and is used to provide an excitation light La to irradiate the plurality of biological particles, and the target biological particles among the plurality of biological particles are irradiated by the excitation light La and excited to generate a radiation light Lb. It should be noted that the wavelength of the excitation light La of the excitation light source 20 includes but is not limited to infrared light, ultraviolet light and visible light; the wavelength of the excitation light Lb includes but is not limited to infrared light, ultraviolet light and visible light.

[0033] The filter spectroscope assembly 30 includes an objective lens 32, a dichroic mirror 34, a filter assembly 36, and a spectroscope 38. The objective lens 32, the dichroic mirror 34, the filter assembly 36, and the spectroscope 38 are sequentially disposed below the detection stage 10 from top to bottom.

[0034] The objective lens 32 of the filter spectroscope group 30 is located on the optical path of the radiated light Lb and is used to focus the at least one target biological particle and enlarge the image. The dichroic mirror 34 is located on the optical path of the excitation light La and is used to reflect the excitation light La so that the excitation light La irradiates the multiple biological particles after reflection. The dichroic mirror 34 in this embodiment is also located on the optical path of the radiated light Lb, and the radiated light Lb can penetrate the dichroic mirror 34. The filter group 36 is located on the optical path of the radiated light Lb and includes a plurality of filters (not shown), wherein the plurality of filters are used to allow light of different wavelengths to pass through, so that the plurality of filters can be replaced according to user needs, and the plurality of filters can be used corresponding to the radiated light Lb of different wavelengths generated by different types of target biological particles, so that light of specific one or more wavelength bands can be penetrated while the remaining wavelength bands are excluded.

[0035] The spectroscope 38 is located on the optical path of the radiation light Lb, and is used to split the incident radiation light Lb into a first detection light L1 and a second detection light L2 that account for different proportions of the radiation light. Under the condition that the energy of the radiation light Lb absorbed by the spectroscope 38 during the transmission process is ignored, the energy of the radiation light Lb is equal to the sum of the energies of the first detection light L1 and the second detection light L2. The spectroscope 38 is usually defined with a transmittance and a reflectance, wherein the sum of the transmittance and the reflectance is 100%. The transmittance of the spectroscope 38 in this embodiment is 10%, and the reflectance is 90%, which means that 10% of the energy of the radiation light Lb received by the spectroscope 38 will be transmitted and the other 90% will be reflected. The aforementioned 10% of the radiation light Lb that transmits the spectroscope 38 is the first detection light L1, and the aforementioned 90% of the radiation light Lb that is reflected from the spectroscope 38 is the second detection light L2. The ratio of the first detection light L1 to the radiated light Lb of the present invention is between 0.1 and 0.5, and the ratio of the second detection light L2 to the radiated light Lb is between 0.5 and 0.9; preferably, the ratio of the first detection light L1 to the radiated light Lb is between 0.1 and 0.3, and the ratio of the second detection light L2 to the radiated light Lb is between 0.7 and 0.9; in this embodiment, the ratio of the first detection light L1 to the radiated light Lb is 0.1, and the ratio of the second detection light L2 to the radiated light Lb is 0.9.

[0036] The scanning sensor group 40 is connected to the spectroscope 38 and includes a photomultiplier tube 42 (PMT) and a charge-coupled device 44 (CCD). The CCD 44 and the filter group 36 are located on the same side of the mirror surface of the spectroscope 38 and are used to receive the second detection light L2, i.e., the reflected light; the PMT 42 is located on the other side of the mirror surface of the spectroscope 38 and is used to receive the first detection light L1, i.e., the transmitted light. However, the present invention can change the reflection and transmission ratio of the spectroscope according to the needs of the user, for example, the PMT 42 and the filter group 36 are located on the same side of the mirror surface of the spectroscope 38 and are used to receive the first detection light L1, i.e., the reflected light; the CCD 44 is located on the other side of the mirror surface of the spectroscope 38 and is used to receive the second detection light L2, i.e., the transmitted light. The photomultiplier tube 42 is a light detection element with high sensitivity and ultra-fast response time, which can quickly scan a large area. When the photomultiplier tube 42 scans the detection stage 10 and receives the first detection light L1, the photomultiplier tube 42 generates a regional positioning signal, which includes the regional position and fluorescence signal intensity of the target biological particle on the detection stage 10. The charge-coupled device 44 can finely detect a digital image of a regional position. When the charge-coupled device 44 scans the regional position on the detection stage 10 and receives the second detection light L2, the charge-coupled device 44 generates an image signal, which includes the image position of the target biological particle on the detection stage 10.

[0037] More specifically, see Figure 1 , Figure 2 and Figure 3 As shown, the detection device 100 performs a detection method X through the above mechanical structure to scan and detect the multiple biological particles on its detection carrier 10. The detection method X includes the following steps A to G:

[0038] Step A: The excitation light source 20 emits excitation light La, and the dichroic mirror 34 reflects the excitation light La and then irradiates the plurality of biological particles.

[0039] Step B: The at least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiation light Lb, and the radiation light Lb is incident on the filter spectroscope assembly 30 .

[0040] Steps between step B and step C: the objective lens 32 of the filter spectroscope group 30 allows the radiation light Lb to enter the filter group 36. Then, the filter group 36 selects a filter that can only transmit the wavelength band of the radiation light Lb, filters out the excitation light La or light of other wavelength bands, and allows the radiation light Lb to enter the spectroscope 38 again.

[0041] Step C: The beam splitter 38 of the filter beam splitter assembly 30 splits the radiation light Lb into the first detection light L1 and the second detection light L2 , and the first detection light L1 and the second detection light L2 are incident on the photomultiplier tube 42 and the charge coupled device 44 respectively.

[0042] Step D: The photomultiplier tube 42 and the detection stage 10 move relative to each other to scan the regional position on the detection stage 10. When the photomultiplier tube 42 receives the first detection light L1 at a regional position of the detection stage 10, the photomultiplier tube 42 will generate a regional positioning signal, which includes the fluorescence signal intensity of the at least one target biological particle on the detection stage 10 and is transmitted to the processor.

[0043] Step E: The processor determines that the regional position on the detection stage 10 has the at least one target biological particle according to the regional positioning signal, and transmits the regional positioning signal to the charge coupled device 44 .

[0044] Step F: The CCD 44 receives the second detection light L2 to sense the image position of the at least one target biological particle in the area, thereby generating an image signal.

[0045] Step G: The processor obtains the precise position of the at least one target biological particle in the regional position on the detection platform 10 according to the image signal, and returns to step D to scan other regional positions on the detection platform 10 that have not been detected.

[0046] When the detection device 100 detects the target biological particles by the detection method X, steps D to G are repeatedly performed until all the regional positions are detected. The advantage of using the detection method X for detection is that, when the number of cells on the sorting disk on the detection stage 10 is small, when the photomultiplier tube 42 detects the target biological particles in any regional position, the charge coupled device 44 can immediately generate an image signal of the regional position, so that the metal needle (not shown) can immediately go to the regional position to absorb the target biological particles, thereby avoiding the target biological particles from staying in the sorting disk for too long, and reducing the problem of cells sticking to the sorting disk and being difficult to absorb.

[0047] For example, the detection platform 10 can be divided into 5 equal-area regional positions, represented as A, B, C, D, and E (not shown in the figure). However, those with ordinary knowledge in the field can divide the regional positions of different areas and different numbers according to actual needs. The multiple biological particles are randomly distributed in the regional positions, so that some regional positions have at least one target biological particle, and some regional positions do not have the target biological particles. The target biological particles can be identified by the user by matching different labels, fluorescence selection, and filter selection. In step D, the photomultiplier tube 42 and the detection platform 10 move relative to each other, and scan the regional positions A, B, C, D, and E on the detection platform 10 respectively to sense at least one target biological particle.

[0048] As mentioned above, when executing the detection method X, when the photomultiplier tube 42 is at the area position A on the detection stage 10 and senses the first detection light L1 generated by the label on at least one target biological particle being excited by the excitation light, the photomultiplier tube 42 will generate a regional positioning signal to record the fluorescence signal intensity of the area position A, and transmit the regional positioning signal information to the processor. Then, the processor determines that the area position A on the detection stage 10 has at least one target biological particle based on the regional positioning signal, and thus transmits the regional positioning signal containing the position information of the area position A to the charge coupled device 44, so that the charge coupled device 44 receives the second detection light L2 to sense the image position of the at least one target biological particle in the area position A, thereby generating an image signal. The processor obtains the precise position of at least one target biological particle in the area position A on the detection stage 10 based on the image signal. The advantage of this method is that when the photomultiplier tube 42 receives the fluorescent signal of the target biological particle, the charge-coupled device 44 can be used to instantly identify the precise position of the target biological particle, and the target biological particle can be instantly separated from the regional position, while preventing the target biological particle from staying on the detection platform 10 for too long, thereby reducing the problem of the target biological particle adhering to the detection platform 10 and being difficult to separate.

[0049] When performing the detection method X, when the photomultiplier tube 42 senses the radiation light Lb generated by the label on at least one target biological particle being excited by the excitation light at the regional position A on the detection stage 10, the photomultiplier tube 42 will generate a regional positioning signal, the regional positioning signal includes the fluorescence signal intensity of the regional position A, and the photomultiplier tube 42 transmits the regional positioning signal to the processor. Then, the processor determines that the regional position A on the detection stage 10 has at least one target biological particle according to the regional positioning signal, and makes the charge-coupled device 44 receive the second detection light L2 to sense the image position of the at least one target biological particle at the regional position A, thereby generating an image signal. The processor obtains the precise position of at least one target biological particle in the region position A on the detection stage 10 according to the generated image signal. Then, when the photomultiplier tube 42 senses the radiated light Lb generated by the label on at least one target biological particle being excited by the excitation light at another region position B on the detection stage 10, the photomultiplier tube 42 will generate another region positioning signal, the region positioning signal includes the fluorescence signal intensity of the region position B, and the photomultiplier tube 42 transmits the another region positioning signal to the processor. Then, the processor determines that the region position B on the detection stage 10 has at least one target biological particle according to the region positioning signal, and causes the charge coupled device 44 to receive the second detection light L2 to sense the image position of the at least one target biological particle in the region position B, thereby generating an image signal. The processor obtains the precise position of at least one target biological particle in the region position B on the detection stage 10 according to the generated image signal.

[0050] Please continue to Figure 1 , Figure 2 and Figure 3 As shown, the detection device 100 can also perform another detection method Y through the above mechanical structure to scan and detect the multiple biological particles on its detection carrier 10. The detection method Y includes the following steps A to G:

[0051] Step A: The excitation light source 20 emits excitation light La, and the dichroic mirror 34 reflects the excitation light La and then irradiates the plurality of biological particles.

[0052] Step B: At least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiation light Lb, which enters the filter spectroscope assembly 30 .

[0053] Steps between step B and step C: the objective lens 32 of the filter spectroscope group 30 allows the radiation light Lb to enter the filter group 36. Then, the filter group 36 selects a filter that can only transmit the wavelength band of the radiation light Lb, filters out the excitation light La or light of other wavelength bands, and allows the radiation light Lb to enter the spectroscope 38 again.

[0054] Step C: The beam splitter 38 of the filter beam splitter assembly 30 splits the radiation light Lb into the first detection light L1 and the second detection light L2 , and the first detection light L1 and the second detection light L2 are incident on the photomultiplier tube 42 and the charge coupled device 44 respectively.

[0055] Step D: The photomultiplier tube 42 and the detection stage 10 move relative to each other to scan multiple regional positions on the detection stage 10. When the photomultiplier tube 42 receives the first detection light L1 at any regional position of the detection stage 10, the photomultiplier tube 42 will generate a regional positioning signal, which includes the regional position and fluorescence signal intensity of the at least one target biological particle on the detection stage 10, and is transmitted to the processor until all regional positions are scanned.

[0056] Step E: The processor determines that at least one of the regional positions on the detection platform 10 has the at least one target biological particle according to at least one of the regional positioning signals, and transmits at least one of the regional positioning signals to the charge coupled device 44 .

[0057] Step F: The charge-coupled device 44 moves relative to the detection platform 10 according to at least one of the regional positioning signals, and receives the second detection light L2 at each of the regional positions to sense the image position of at least one target biological particle on the detection platform 10, thereby generating an image signal respectively.

[0058] Step G: The processor obtains the precise position of the at least one target biological particle in at least one of the regional positions on the detection stage 10 according to at least one of the image signals.

[0059] When the detection device 100 detects target biological particles by the above detection method Y, since the photomultiplier tube 42 performs detection method Y to scan all regional positions at once, it is not necessary to repeatedly stop after scanning a regional position, as the photomultiplier tube 42 performs detection method X, and then scan another regional position until other components are completed, so the scanning speed of detection method Y is faster than that of detection method X. In some cases where the number of cells in the sorting disk on the detection platform 10 is small, the total processing time of the detection device 100 for detection by detection method X and detection method Y is similar. However, in some cases where the number of cells in the sorting disk on the detection platform 10 is large, detection efficiency can be improved by using detection method Y for detection.

[0060] For example, the detection platform 10 can be divided into 5 equal-area regional positions, represented as A, B, C, D, and E (not shown in the figure). However, those with ordinary knowledge in the field can divide the regional positions of different areas and different numbers according to actual needs. The multiple biological particles are randomly distributed in the regional positions, so that some regional positions have at least one target biological particle, and some regional positions do not have the target biological particles. The target biological particles can be identified by the user by matching different labels, fluorescence selection, and filter selection. In step D, the photomultiplier tube 42 and the detection platform 10 move relative to each other, and scan the regional positions A, B, C, D, and E on the detection platform 10 respectively to sense at least one target biological particle.

[0061] As mentioned above, when executing the detection method Y, when the photomultiplier tube 42 senses the radiation light Lb generated by the label on at least one target biological particle being excited by the excitation light at the regional position A on the detection stage 10, the photomultiplier tube 42 will generate a regional positioning signal to record the position and fluorescence signal intensity of the regional position A, and transmit the regional positioning signal information to the processor, and the photomultiplier tube 42 continues to move to other regional positions B, C, D and E on the detection stage 10 until all regional positions are scanned. Then, the processor determines that the regional position A on the detection stage 10 has at least one target biological particle based on the regional positioning signal, and thus transmits the regional positioning signal containing the position information of the regional position A to the charge coupled device 44. The charge coupled device 44 moves to the regional position A relative to the detection stage 10 based on the regional positioning signal, and receives the second detection light L2 to sense the image position of the at least one target biological particle at the regional position A, thereby generating an image signal. The processor obtains the precise position of at least one target biological particle in the area position A on the detection stage 10 according to the image signal. The advantage of this method is that the photomultiplier tube 42 is used to quickly complete the scanning of all area positions, and then the charge-coupled device 44 is used to confirm whether there are target biological particles in the field of view for the area position with fluorescent signals, thereby improving the efficiency of scanning detection.

[0062] As mentioned above, when executing the detection method Y, when the photomultiplier tube 42 senses the radiation light Lb generated by the label on at least one target biological particle being excited by the excitation light at the regional positions A and B on the detection stage 10, the photomultiplier tube 42 will generate two regional positioning signals, the two regional positioning signals respectively including the position information and fluorescence signal intensity of the regional positions A and B, and the photomultiplier tube 42 transmits the two regional positioning signals to the processor, and the photomultiplier tube 42 continues to move to other regional positions C, D and E on the detection stage 10 until all regional positions are scanned. Then, the processor determines that the regional positions A and B on the detection stage 10 respectively have at least one target biological particle based on the two regional positioning signals, and thus transmits the regional positioning signal including the position information of the regional positions A and B to the charge coupled device 44. The charge coupled device 44 moves to the regional positions A and B relative to the detection stage 10 respectively according to the two regional positioning signals, and respectively receives the second detection light L2 to sense the image position of the at least one target biological particle in the regional positions A and B on the detection stage 10, thereby generating an image signal respectively. The processor obtains the precise position of the at least one target biological particle in the regional positions A and B on the detection stage 10 respectively according to the two generated image signals.

[0063] The optical system of the detection device 100 forms a first optical path S1 ( Figure 2 ) and a second optical path S2 ( Figure 3 ), thereby knowing the precise location of at least one target biological particle mentioned above.

[0064] Reference Figure 2 The first optical path S1 starts from the excitation light source 20, passes through the dichroic mirror 34, the objective lens 32, the detection stage 10, the objective lens 32, the dichroic mirror 34, the filter set 36 in sequence, and then penetrates the beam splitter 38 to reach the photomultiplier tube 42. After receiving the first detection light L1 transmitted on the first optical path S1, the photomultiplier tube 42 can quickly determine which area positions on the detection stage 10 contain the at least one target biological particle, so as to obtain the area position of the at least one target biological particle.

[0065] Reference Figure 3 The second optical path S2 starts from the excitation light source 20, passes through the dichroic mirror 34, the objective lens 32, the detection stage 10, the objective lens 32, the dichroic mirror 34, the filter set 36 in sequence, and is reflected by the beam splitter 38 to reach the charge-coupled device 44. After the charge-coupled device 44 receives the second detection light L2 transmitted on the second optical path S2, it can accurately obtain the precise position of the at least one target biological particle. In this way, the processor determines the precise position of the at least one target biological particle on the detection stage 10 through the photomultiplier tube and the charge-coupled device, and then the processor can control other devices to select the at least one target biological particle.

[0066] It is supplemented that, although in the present embodiment, the ratio of the first detection light L1 to the radiated light Lb is 0.1, and the ratio of the second detection light L2 to the radiated light Lb is 0.9, in other practices, a spectroscope with different ratios of transmittance and reflectance may be used. For example, a spectroscope with a transmittance of 20% and a reflectance of 80% may be used, in which the ratio of the first detection light L1 to the radiated light Lb is 0.2, and the ratio of the second detection light L2 to the radiated light Lb is 0.8; or, a spectroscope with a transmittance of 50% and a reflectance of 50% may be used, in which the ratio of the first detection light L1 to the radiated light Lb is 0.5, and the ratio of the second detection light L2 to the radiated light Lb is 0.5, but is not limited to the above.

[0067] In this embodiment, another embodiment of the beam splitter 38 and the scanning sensor set 40 is provided. Figure 4As shown, the transmittance of the beam splitter 38 is 90% and the reflectance is 10%, wherein the 10% radiated light Lb reflected from the beam splitter 38 is the first detection light L1, and the 90% radiated light Lb that penetrates the beam splitter 38 is the second detection light L2. Similarly, the photomultiplier tube 42 of the scanning sensor group 40 receives the first detection light L1, and the charge coupled device 44 receives the second detection light L2.

[0068] Please refer to Figure 5 As shown, the detection device 100A of the second preferred embodiment of the present invention is different from the detection device 100 of the first preferred embodiment in that no filter set is set between the dichroic mirror 34A and the beam splitter 38A of the detection device 100A, but the optical system of the detection device 100A includes a first filter set 36A and a second filter set 37A. The beam splitter 38A is located on the optical path of the radiated light Lb, and splits the radiated light Lb into the first detection light L1 and the second detection light L2 for emission. The first filter set 36A and the second filter set 37A are respectively located on the optical paths of the first detection light L1 and the second detection light L2. In this way, compared with the detection method X or Y of the first preferred embodiment when executing (step C), after the beam splitter 38A of the second preferred embodiment splits the radiated light Lb into the first detection light L1 and the second detection light L2 for emission, the detection method will perform the following steps between step C and step D:

[0069] The first filter group 36A uses a filter that can only transmit the wavelength of the first detection light L1, and filters out the excitation light La or light of other wavelengths before entering the photomultiplier tube 42A; and

[0070] The second filter group 37A uses a filter that only allows the wavelength band of the second detection light L2 to pass through, and filters out the excitation light La or light in other wavelength bands before it enters the charge coupled device 44A.

[0071] As can be seen from the above, the first optical path received by the photomultiplier tube 42A starts from the excitation light source 20A, and sequentially passes through the dichroic mirror 34A, the objective lens 32A, the detection stage 10A, the objective lens 32A, the dichroic mirror 34A, and penetrates through the beam splitter 38A, and then after being filtered by the first filter set 36A, it reaches the photomultiplier tube 42A. The second optical path received by the charge-coupled device 44A starts from the excitation light source 20A, sequentially passes through the dichroic mirror 34A, the objective lens 32A, the detection stage 10A, the objective lens 32A, the dichroic mirror 34A, and after being reflected by the beam splitter 38A, and then after being filtered by the second filter set 37A, it reaches the charge-coupled device 44A. The above first and second filter sets 36A, 37A can be replaced according to the user's needs, and specific one or more bands of light can be made to penetrate while the remaining bands are excluded according to the needs. The advantage is that the photomultiplier tube 42A and the charge-coupled device 44A can simultaneously view different target biological particles according to the user's needs.

[0072] When actually using the detection device 100A of this embodiment, when the photomultiplier tube 42A receives the first detection light L1 at a regional position on the detection stage 10, and the charge-coupled device 44A receives the second detection light L2, the filter of the second filter set 37A can be replaced with a filter capable of detecting other fluorescent particle labels. In this way, it can be used to observe whether there are target biological particles combined with another label at the regional position. Therefore, the photomultiplier tube 42A and the charge-coupled device 44A can simultaneously view different target biological particles, which helps to reduce the time for switching filters.

[0073] Please refer to Figure 6 As shown, it is the detection device 100B of the third preferred embodiment of the present invention. The difference between the detection device 100B of the third preferred embodiment and the detection device 100 of the first preferred embodiment is that the placement positions of the components in the filter and beam splitter group of the detection device 100B are different from the placement positions of the components in the filter and beam splitter group 30 of the detection device 100, thereby affecting the placement positions of the excitation light source 20B, the photomultiplier tube 42B, and the charge-coupled device 44B.

[0074] The objective lens 32B and the dichroic mirror 34B of the filter and dichroic mirror group of the detection device 100B are arranged in sequence up and down below the detection stage 10B, and the filter group 36B and the dichroic mirror 38B are arranged on the same side in front of the dichroic mirror 34B, and the filter group 36B is located between the dichroic mirror 34B and the dichroic mirror 38B. The excitation light source 20B is arranged below the dichroic mirror 34B. The photomultiplier tube 42B is disposed below the spectroscope 38B, and the charge-coupled device 44B is disposed on the side of the spectroscope 38B away from the filter group 36B. However, since the ratio of the transmittance and reflectance of the spectroscope 38B varies depending on the usage scenario, the spectroscope, photomultiplier tube and charge-coupled device of the present invention can be used in combination according to the following principles: the spectroscope of the present invention will divide the radiated light into a transmitted light and a reflected light, wherein the light with lower energy between the transmitted light and the reflected light serves as the first detection light of the present invention, and the light with higher energy between the transmitted light and the reflected light serves as the second detection light of the present invention, and the photomultiplier tube constantly receives the first detection light, and the charge-coupled device constantly receives the second detection light.

[0075] The detection method X of the detection device 100B of the third preferred embodiment comprises the following steps A to G:

[0076] Step A: The excitation light source 20B emits an excitation light La, and the excitation light La penetrates the dichroic mirror 34B and the objective lens 32B to irradiate the plurality of biological particles.

[0077] Step B: At least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiation light Lb, which enters the filter spectroscope assembly.

[0078] Steps between step B and step C: the objective lens 32B of the filter spectroscope group makes the radiation light Lb incident on the dichroic mirror 34B, and the dichroic mirror 34B reflects the radiation light Lb to the filter group 36B. Then, the filter group 36B selects a filter that can only transmit the wavelength band of the radiation light Lb, filters out the excitation light La or light of other wavelength bands, and makes the radiation light Lb incident on the spectroscope 38B again.

[0079] Step C: The beam splitter 38B of the filter beam splitter group splits the radiated light Lb into the first detection light L1 and the second detection light L2, and the first detection light L1 and the second detection light L2 are incident on the photomultiplier tube 42B and the charge coupled device 44B respectively.

[0080] Step D: The photomultiplier tube 42B and the detection stage 10B move relative to each other to scan the regional position on the detection stage 10B. When the photomultiplier tube 42B receives the first detection light L1 at a regional position of the detection stage 10B, the photomultiplier tube 42B will generate a regional positioning signal, which includes the fluorescence signal intensity of the at least one target biological particle on the detection stage 10 and is transmitted to the processor.

[0081] Step E: The processor determines that the regional position on the detection stage 10B has the at least one target biological particle according to the regional positioning signal, and transmits the regional positioning signal to the charge coupled device 44B.

[0082] Step F: The CCD 44B receives the second detection light L2 to sense the image position of the at least one target biological particle at the regional position, thereby generating an image signal.

[0083] Step G: The processor obtains the precise position of the at least one target biological particle in the regional position on the detection platform 10B according to the image signal, and returns to step D to scan other regional positions on the detection platform 10B that have not been detected.

[0084] Another detection method Y of the detection device 100B of the third preferred embodiment includes the following steps A to G:

[0085] Step A: The excitation light source 20B emits an excitation light La, and the excitation light La penetrates the dichroic mirror 34B and the objective lens 32B to irradiate the plurality of biological particles.

[0086] Step B: At least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiated light Lb, which enters the filter spectroscope assembly.

[0087] Steps between step B and step C: the objective lens 32B of the filter spectroscope group makes the radiation light Lb incident on the dichroic mirror 34B, and the dichroic mirror 34B reflects the radiation light Lb to the filter group 36B. Then, the filter group 36B selects a filter that can only transmit the wavelength band of the radiation light Lb, filters out the excitation light La or light of other wavelength bands, and makes the radiation light Lb incident on the spectroscope 38B again.

[0088] Step C: The beam splitter 38B of the filter beam splitter group splits the radiated light Lb into the first detection light L1 and the second detection light L2, and the first detection light L1 and the second detection light L2 are incident on the photomultiplier tube 42B and the charge coupled device 44B respectively.

[0089] Step D: The photomultiplier tube 42B and the detection stage 10B move relative to each other to scan multiple regional positions on the detection stage 10B. When the photomultiplier tube 42B receives the first detection light L1 at any regional position of the detection stage 10B, the photomultiplier tube 42B will generate a regional positioning signal, which includes the regional position and fluorescence signal intensity of the at least one target biological particle on the detection stage 10B, and is transmitted to the processor until all regional positions are scanned.

[0090] Step E: The processor determines that at least one of the regional positions on the detection platform 10B has the at least one target biological particle according to at least one of the regional positioning signals, and transmits at least one of the regional positioning signals to the charge coupled device 44B.

[0091] Step F: The charge-coupled device 44B moves relative to the detection platform 10B according to at least one of the regional positioning signals, and receives the second detection light L2 at each of the regional positions to sense the image position of at least one target biological particle on the detection platform 10B, thereby generating an image signal respectively.

[0092] Step G: The processor obtains the precise position of the at least one target biological particle in at least one of the regional positions on the detection platform 10B according to at least one of the image signals.

[0093] The above structural relationship makes the detection device 100B of the third preferred embodiment have a lower overall device height and a larger width compared to the detection device 100 of the first preferred embodiment, and is more suitable for placement in a shorter and wider storage space. Purchasers can choose according to usage requirements.

[0094] Please refer to Figure 7 As shown, a detection device 100C according to a fourth preferred embodiment of the present invention is different from the detection device 100B according to the third preferred embodiment in that no filter group is arranged between the color dichroic mirror 34C and the beam splitter 38C of the detection device 100C, but the filter beam splitter group of the detection device 100C includes a first filter group 36C and a second filter group 37C. The beam splitter 38C is located on the optical path of the radiated light Lb, and the first filter group 36C and the second filter group 37C are respectively located on the optical paths of the first detection light L1 received by the photomultiplier tube 42C and the second detection light L2 received by the charge coupled device 44C.

[0095] Thus, the steps of the detection method of the detection device 100C of the fourth preferred embodiment are substantially the same as those of the detection method of the detection device 100A of the second preferred embodiment. The difference is that the detection method of the detection device 100C of the fourth preferred embodiment is compared with step A of the second preferred embodiment. The fourth preferred embodiment performs the following steps in step A:

[0096] The excitation light source 20C emits excitation light La, and the excitation light La penetrates the dichroic mirror 34C and the objective lens 32C, and then irradiates the plurality of biological particles on the detection stage 10C.

[0097] The first and second filter sets 36C and 37C can be replaced according to user needs to allow light of one or more specific wavelength bands to pass through while excluding the rest of the wavelength bands. The advantage is that the photomultiplier tube 42C and the charge-coupled device 44C can simultaneously inspect different types of target biological particles according to user needs.

[0098] When the detection device 100C of this embodiment is actually used, the photomultiplier tube 42C receives the first detection light L1 at a regional position on the detection stage 10, and after the charge-coupled device 44C receives the second detection light L2, the filter of the second filter group 37C can be replaced with a filter that can detect labels of other fluorescent particles. In this way, it can be used to observe whether the regional position has target biological particles combined with another label. Therefore, the photomultiplier tube 42C and the charge-coupled device 44C can sense and inspect different types of target biological particles at the same time, which helps to reduce the time of switching filters. In addition, the detection device 100C of the fourth preferred embodiment has the same advantages as the detection device 100B of the third preferred embodiment, and is more suitable for placement in a shorter and wider accommodation space.

[0099] Please refer to Figure 8 As shown, it is a detection device 100D of the fifth preferred embodiment of the present invention. The difference between the detection device 100D of the fifth preferred embodiment and the detection device 100 of the first preferred embodiment is that the excitation light source 20D and the objective lens 32D are arranged on different sides of the detection carrier 10D, and the detection carrier 10D is located on the light path of the excitation light La of the excitation light source 20D, so that the excitation light La is directly irradiated on the multiple biological particles. Therefore, the filter spectroscope group of the detection device 100D does not need to be provided with a color separation mirror for reflecting the excitation light La, wherein the objective lens 32D of the filter spectroscope group, the filter group 36D and the spectroscope 38D are arranged in order from top to bottom below the detection carrier 10D.

[0100] The detection method X of the detection device 100D comprises the following steps A to G:

[0101] Step A: The excitation light source 20D emits excitation light La, and the excitation light La directly irradiates the plurality of biological particles.

[0102] Step B: The at least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiated light Lb, which enters the filter spectroscope assembly 30D.

[0103] Steps between step B and step C: the objective lens 32D of the filter spectroscope group 30D allows the radiation light Lb to enter the filter group 36D. Then, the filter group 36D selects a filter that can only transmit the wavelength band of the radiation light Lb, filters out the excitation light La or light of other wavelength bands, and allows the radiation light Lb to enter the spectroscope 38D again.

[0104] Step C: The beam splitter 38D splits the radiation light Lb into the first detection light L1 and the second detection light L2, and the first detection light L1 and the second detection light L2 are incident on the photomultiplier tube 42D and the charge coupled device 44D respectively.

[0105] Step D: The photomultiplier tube 42D and the detection stage 10D move relative to each other to scan the regional position on the detection stage 10D. When the photomultiplier tube 42D receives the first detection light L1 at a regional position of the detection stage 10D, the photomultiplier tube 42D generates a regional positioning signal, which includes the fluorescence signal intensity of the at least one target biological particle on the detection stage 10D and is transmitted to the processor.

[0106] Step E: The processor determines that the regional position on the detection stage 10D has the at least one target biological particle according to the regional positioning signal, and transmits the regional positioning signal to the charge coupled device 44D.

[0107] Step F: The CCD 44D receives the second detection light L2 to sense the image position of the at least one target biological particle at the regional position, thereby generating an image signal.

[0108] Step G: The processor obtains the precise position of the at least one target biological particle in the regional position on the detection platform 10D according to the image signal, and returns to step D to scan other regional positions on the detection platform 10D that have not been detected.

[0109] The detection method Y of the detection device 100D comprises the following steps A to G:

[0110] Step A: The excitation light source 20D emits excitation light La, and the excitation light La directly irradiates the plurality of biological particles.

[0111] Step B: The at least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiated light Lb, which enters the filter spectroscope assembly 30D.

[0112] Steps between step B and step C: the objective lens 32D of the filter spectroscope group 30D allows the radiation light Lb to enter the filter group 36D. Then, the filter group 36D selects a filter that can only transmit the wavelength band of the radiation light Lb, filters out the excitation light La or light of other wavelength bands, and allows the radiation light Lb to enter the spectroscope 38D again.

[0113] Step C: The beam splitter 38D splits the radiation light Lb into the first detection light L1 and the second detection light L2, and the first detection light L1 and the second detection light L2 are incident on the photomultiplier tube 42D and the charge coupled device 44D respectively.

[0114] Step D: The photomultiplier tube 42D and the detection stage 10D move relative to each other to scan multiple regional positions on the detection stage 10D. When the photomultiplier tube 42D receives the first detection light L1 at any regional position of the detection stage 10D, the photomultiplier tube 42D will generate a regional positioning signal, which includes the regional position and fluorescence signal intensity of the at least one target biological particle on the detection stage 10D, and is transmitted to the processor until all regional positions are scanned.

[0115] Step E: The processor determines that at least one of the regional positions on the detection platform 10D has the at least one target biological particle according to at least one of the regional positioning signals, and transmits at least one of the regional positioning signals to the charge coupled device 44D.

[0116] Step F: The charge-coupled device 44D moves relative to the detection platform 10D according to at least one of the regional positioning signals, and receives the second detection light L2 at each of the regional positions to sense the image position of at least one target biological particle on the detection platform 10D, thereby generating an image signal respectively.

[0117] Step G: The processor obtains the precise position of the at least one target biological particle in at least one of the regional positions on the detection stage 10D according to at least one of the image signals.

[0118] The advantages of the detection device 100D of the fifth preferred embodiment are that: the filter spectroscope set is not arranged behind the dichroic mirror, which can reduce the influence on the radiated light and avoid the energy loss of the radiated light and the decrease of the scanning sensing quality. In addition, the excitation light source 20D, the detection stage 10D and the filter spectroscope set 30D of the detection device 100D of the fifth preferred embodiment can all move relative to each other, thereby increasing the free mobility of the machine.

[0119] Please refer to Fig. 9 1 is a detection device 100E of the sixth preferred embodiment of the present invention. The difference between the detection device 100E of the sixth preferred embodiment and the detection device 100D of the fifth preferred embodiment is that the filter spectroscope group of the detection device 100E includes an objective lens 32E, three dichroic mirrors 34E, a filter group 36E and a spectroscope 38E, and the scanning sensor group of the detection device 100E includes four photomultiplier tubes 42E and a charge coupled device 44E. The objective lens 32E and the spectroscope 38E are arranged in sequence up and down below the detection stage 10E, and the spectroscope 34E is arranged below the spectroscope 38E and located on the optical path of the first detection light L1, so as to split the incident first detection light L1 into a plurality of first detection light bands L11 of different wavelengths and emit them. Another two dichroic mirrors 34E are arranged on one side of the aforementioned dichroic mirror 34E, and the two dichroic mirrors 34E receive at least one of the multiple first detection light bands L11, thereby the multiple photomultiplier tubes 42E can respectively generate a regional positioning signal, and the multiple regional positioning signals can respectively represent the regional positions of different types of target biological particles on the detection carrier 10E.

[0120] For example, the biological particles on the detection carrier 10E include a plurality of different target biological particles, and one of the plurality of different target biological particles can be combined with a plurality of different labels at the same time, or the plurality of different target biological particles are respectively combined with different labels. The user can excite the plurality of labels on the detection carrier 10E by replacing the excitation light emitted by the excitation light source, and the plurality of labels can respectively emit different radiation lights after being excited. When the plurality of photomultiplier tubes 42E respectively receive different radiation lights, they can respectively generate a regional positioning signal, which represents that one of the target biological particles in a certain regional position on the detection carrier 10E may be combined with a plurality of different fluorescent particle labels at the same time or a plurality of different target biological particles are respectively combined with different fluorescent particle labels. In this way, during the scanning detection process, it is possible to simultaneously check whether different fluorescent labels exist in a certain regional position on the detection carrier 10E.

[0121] The detection method X of the detection device 100E of the sixth preferred embodiment includes the following steps A to G:

[0122] Step A: The excitation light source 20E emits excitation light La, and the excitation light La directly irradiates the plurality of biological particles.

[0123] Step B: At least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiated light Lb, which enters the filter spectroscope assembly.

[0124] The step between step B and step C: the objective lens 32E of the filter spectroscope assembly allows the radiation light Lb to enter the spectroscope 38E.

[0125] Step C: The spectroscope 38E splits the radiated light Lb into the first detection light L1 and the second detection light L2, and the multiple different first detection light bands L11 of the first detection light L1 are incident on the multiple photomultiplier tubes 42E respectively; the filter group allows the band of the second detection light L2 to pass through the filter, filters out the excitation light La or light in other bands, and allows the radiated light Lb to be incident on the charge-coupled device 44E again.

[0126] Step D: The plurality of photomultiplier tubes 42E and the detection stage 10E move relative to each other to scan the regional position on the detection stage 10E. When the photomultiplier tube 42E receives the first detection light L1 at a regional position of the detection stage 10E, the photomultiplier tube 42E will generate a regional positioning signal, which includes the fluorescence signal intensity of the at least one target biological particle on the detection stage 10, and is transmitted to the processor.

[0127] Step E: The processor determines that the regional position on the detection platform 10E has the target biological particles according to the regional positioning signal, and transmits the regional positioning signal to the charge coupled device 44E.

[0128] Step F: The CCD 44E receives the second detection light L2 to sense the image position of the at least one target biological particle at the regional position, thereby generating at least one image signal.

[0129] Step G: The processor obtains the precise position of the at least one target biological particle in the regional position on the detection platform 10E according to the image signal, and returns to step D to scan other regional positions on the detection platform 10E that have not been detected.

[0130] Another detection method Y of the detection device 100E of the sixth preferred embodiment includes the following steps A to G:

[0131] Step A: The excitation light source 20E emits excitation light La, and the excitation light La directly irradiates the plurality of biological particles.

[0132] Step B: The at least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiation light Lb, and the radiation light Lb is incident on the filter spectroscope assembly.

[0133] The step between step B and step C: the objective lens 32E of the filter spectroscope assembly allows the radiation light Lb to enter the spectroscope 38E.

[0134] Step C: The spectroscope 38E splits the radiated light Lb into the first detection light L1 and the second detection light L2, and the multiple different first detection light bands L11 of the first detection light L1 are incident on the multiple photomultiplier tubes 42E respectively; the filter group allows the band of the second detection light L2 to pass through the filter, filters out the excitation light La or light in other bands, and allows the radiated light Lb to be incident on the charge-coupled device 44E again.

[0135] Step D: The plurality of photomultiplier tubes 42E and the detection stage 10E move relative to each other to scan a plurality of regional positions on the detection stage 10E. When the photomultiplier tube 42E receives the first detection light L1 at any regional position of the detection stage 10E, the photomultiplier tube 42E will generate a regional positioning signal, which includes the regional position and fluorescence signal intensity of the at least one target biological particle on the detection stage 10E, and is transmitted to the processor until all regional positions are scanned.

[0136] Step E: The processor determines that at least one regional position on the detection platform 10E has at least one target biological particle based on at least one regional positioning signal, and transmits at least one regional positioning signal to the charge coupled device 44E.

[0137] Step F: The charge-coupled device 44E moves relative to the detection platform 10E according to at least one of the regional positioning signals, and receives the second detection light L2 at each of the regional positions to sense the image position of at least one target biological particle on the detection platform 10E, thereby generating an image signal respectively.

[0138] Step G: The processor obtains the precise position of the at least one target biological particle in at least one of the regional positions on the detection stage 10E according to the image signal.

[0139] Please refer to Fig.10As shown, the detection device 100F of the seventh preferred embodiment of the present invention is different from the detection device 100E of the sixth preferred embodiment in that the filter spectroscope group of the detection device 100F includes an objective lens 32F, five dichroic mirrors 34F and a spectroscope 38F. The objective lens 32F, one of the dichroic mirrors 34F, the spectroscope 38F and another dichroic mirror 34F are arranged in sequence below the detection stage 10F. The dichroic mirror 34F arranged above the spectroscope 38F is used to reflect the excitation light La of the excitation light source 20F so that the excitation light La irradiates the plurality of biological particles on the detection stage 10F. The dichroic mirror 34F arranged below the spectroscope 38F is located on the optical path of the first detection light L1, and is used to split the incident first detection light L1 into a plurality of first detection light bands L11 of different wavelength bands and emit them. Another three dichroic mirrors 34F are arranged on one side of the aforementioned dichroic mirror 34F, and the three dichroic mirrors 34F receive at least one of the multiple first detection light bands L11, thereby the multiple photomultiplier tubes 42F can respectively generate a regional positioning signal, and the multiple regional positioning signals can respectively represent the regional positions of different types of target biological particles on the detection carrier 10F.

[0140] For example, the biological particles on the detection carrier 10F include a plurality of different target biological particles, and one of the plurality of different target biological particles can be combined with a plurality of different labels at the same time, or the plurality of different target biological particles are respectively combined with different labels. The user can excite the plurality of labels on the detection carrier 10F by replacing the excitation light emitted by the excitation light source, and the plurality of labels can respectively emit different radiation lights after being excited. When the plurality of photomultiplier tubes 42F respectively receive different radiation lights, they can respectively generate a regional positioning signal, which represents that one of the target biological particles in a certain regional position on the detection carrier 10F may be combined with a plurality of different fluorescent particle labels at the same time or a plurality of different target biological particles are respectively combined with different fluorescent particle labels. In this way, during the scanning detection process, it is possible to simultaneously check whether different fluorescent labels exist in a certain regional position on the detection carrier 10F.

[0141] The charge coupled element 44F and the filter group 36F of the detection device 100F of the seventh preferred embodiment are arranged on the same side in front of the spectroscope 38F. The filter group 36F allows the wavelength band of the second detection light L2 to penetrate and filters out the excitation light La or light in other wavelength bands. The second detection light L2 is incident on the charge coupled element 44F.

[0142] The above design enables the detection device 100F of the seventh preferred embodiment to save space above the detection stage 10F compared to the detection device 100E of the sixth preferred embodiment, and at the same time provides more photomultiplier tubes 42F to obtain whether different types of target biological particles exist in a specific area position on the detection stage 10F. It can be seen that the number of photomultiplier tubes of the present invention and the number of corresponding dichroic mirrors can be increased or decreased according to usage requirements.

[0143] Please refer to Fig.11 As shown, it is a detection device 100G of the eighth preferred embodiment of the present invention. The difference between the detection device 100G of the eighth preferred embodiment and the detection device 100F of the seventh preferred embodiment is that the filter spectroscope group of the detection device 100G does not include a filter group. Instead, the filter spectroscope group of the detection device 100G is additionally provided with two dichroic mirrors 34G, wherein one dichroic mirror 34G is arranged on one side of the spectroscope 38G and is located on the optical path of the second detection light L2, so as to divide the incident second detection light L2 into a plurality of second detection light bands L21 of different wavelengths and emit them. Another dichroic mirror 34G is arranged on the right side of the aforementioned dichroic mirror 34G, and the dichroic mirror 34G receives at least one of the plurality of second detection light bands L21. The scanning sensor group of the detection device 100G has a plurality of charge-coupled devices 44G, which receive the second detection light band L21 of different bands and respectively generate an image signal. The plurality of image signals can respectively represent the image positions of different types of target biological particles on the detection carrier 10G.

[0144] For example, the biological particles on the detection carrier 10G include multiple different target biological particles, and one of the multiple different target biological particles can be combined with multiple different labels at the same time, or the multiple different target biological particles are respectively combined with different labels. The user can excite the multiple labels on the detection carrier 10G by replacing the excitation light emitted by the excitation light source, and the multiple labels can emit different radiation lights after being excited. When the multiple photomultiplier tubes 42G receive different radiation lights respectively, they can respectively generate a regional positioning signal, which represents that one of the target biological particles in a certain regional position on the detection carrier 10G may be combined with multiple different fluorescent particle labels at the same time or multiple different target biological particles are respectively combined with different fluorescent particle labels. In this way, during the scanning detection process, it is possible to simultaneously check whether different fluorescent labels exist in a certain regional position on the detection carrier 10G. During the scanning detection process, the multiple image signals allow the user to simultaneously check the clear image positions of different types of target biological particles on the detection carrier 10G on multiple different display screens. In this way, the workload of each CCD 44G can be reduced and the effect of receiving different fluorescent label images in a specific field of view can be achieved.

[0145] In some cases, the filter spectroscope assembly of the detection device of the present invention does not include a spectroscope and can also achieve the purpose of simultaneously obtaining the precise position of the target biological particle using a photomultiplier tube and a charge coupled device. Fig.12 As shown, the detection device 100H of the ninth embodiment of the present invention includes a detection carrier 10H, an optical system (not shown) and a processor (not shown) electrically connected to the optical system, wherein the optical system includes an excitation light source 20H, a filter spectrometer group and a scanning sensor group.

[0146] When the excitation light La of the excitation light source 20H irradiates the multiple biological particles, and at least one target biological particle among the multiple biological particles is irradiated by the excitation light La and excited to produce a radiation light Lb, the radiation light Lb will be divided into a first detection light L1 and a second detection light L2 to be emitted, wherein the first detection light L1 and the second detection light L2 have different traveling directions, and the first detection light L1 is opposite to the traveling direction of the excitation light La, and the second detection light L2 is the same as the traveling direction of the excitation light La.

[0147] The filter spectroscope group includes an objective lens 32H, a dichroic mirror 34H, a first filter group 36H and a second filter group 37H. The objective lens 32H and the second filter group 37H are arranged in order from top to bottom below the detection platform 10H, the dichroic mirror 34H is arranged above the detection platform 10H, and the first filter group 36H is located on one side of the dichroic mirror 34H. The objective lens 32H of the filter spectroscope group is located on the optical path of the second detection light L2, and is used to focus the at least one target biological particle and magnify the image. The dichroic mirror 34H is located on the optical path of the first detection light L1 and is used to reflect the first detection light L1. The first filter group 36H and the second filter group 37H are respectively located on the optical paths of the first detection light L1 and the second detection light L2.

[0148] The scanning sensor group includes a photomultiplier tube 42H and a charge-coupled device 44H. The charge-coupled device 44H is located below the second filter group 37H on the optical path of the second detection light L2, and is used to receive the second detection light L2. The photomultiplier tube 42H is located on one side of the filter group 36H on the optical path of the first detection light L1, and is used to receive the first detection light L1. When the photomultiplier tube 42H scans the detection stage 10H relative to the detection stage 10H and receives the first detection light L1, the photomultiplier tube 42H generates a regional positioning signal, and the regional positioning signal includes the regional position of the at least one target biological particle on the detection stage 10H. When the charge-coupled device 44H moves relative to the detection stage 10H and receives the second detection light L2, the charge-coupled device 44H generates an image signal, and the image signal includes the image position of the at least one target biological particle on the detection stage 10H.

[0149] The detection method X of the detection device 100H comprises the following steps A to G:

[0150] Step A: The excitation light source 20H emits excitation light La, and the excitation light La irradiates the plurality of biological particles on the detection stage 10H.

[0151] Step B: The at least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiation light Lb.

[0152] Step C: The radiated light Lb is divided into a first detection light L1 with an upward direction and a second detection light L2 with a downward direction.

[0153] The step between step C and step D: after the dichroic mirror 34H reflects the first detection light L1, it enters the first filter group 36H. The first filter group 36H allows the wavelength band of the first detection light L1 to pass through the filter, filters out the excitation light La or the light of other wavelength bands, and allows the radiation light Lb to enter the photomultiplier tube 42H again; the objective lens 32 allows the second detection light L2 to enter a second filter group 37H. The second filter group 37H allows the wavelength band of the second detection light L2 to pass through the filter, filters out the excitation light La or the light of other wavelength bands, and allows the radiation light Lb to enter the charge coupled device 44H again.

[0154] Step D: The photomultiplier tube 42H and the detection stage 10H move relative to each other to scan the regional position on the detection stage 10H. When the photomultiplier tube 42H receives the first detection light L1 at any regional position of the detection stage 10H, the photomultiplier tube 42H will generate a regional positioning signal, which includes the fluorescence signal intensity of the at least one target biological particle on the detection stage 10H and is transmitted to the processor.

[0155] Step E: The processor determines that the regional position on the detection carrier 10H has the target biological particles according to the regional positioning signal, and transmits the regional positioning signal to the charge coupled device 44H.

[0156] Step F: The processor enables the CCD 44 to receive the second detection light L2 to sense the image position of the at least one target biological particle at the regional position, thereby generating at least one image signal.

[0157] Step G: The processor obtains the precise position of at least one target biological particle in the regional position on the detection platform 10H based on at least one of the image signals, and returns to step D to scan other regional positions on the detection platform 10H that have not been detected.

[0158] Another detection method Y of the detection device 100H comprises the following steps A to G:

[0159] Step A: The excitation light source 20H emits excitation light La, and the excitation light La irradiates the plurality of biological particles on the detection stage 10H.

[0160] Step B: The at least one target biological particle among the plurality of biological particles absorbs the excitation light La to generate radiation light Lb.

[0161] Step C: The radiated light Lb is divided into a first detection light L1 with an upward direction and a second detection light L2 with a downward direction.

[0162] The step between step C and step D: after the dichroic mirror 34H reflects the first detection light L1, it enters the first filter group 36H. The first filter group 36H allows the wavelength band of the first detection light L1 to pass through the filter, filters out the excitation light La or the light of other wavelength bands, and allows the radiation light Lb to enter the photomultiplier tube 42H again; the objective lens 32 allows the second detection light L2 to enter a second filter group 37H. The second filter group 37H allows the wavelength band of the second detection light L2 to pass through the filter, filters out the excitation light La or the light of other wavelength bands, and allows the radiation light Lb to enter the charge coupled device 44H again.

[0163] Step D: The photomultiplier tube 42H and the detection stage 10H move relative to each other to scan multiple regional positions on the detection stage 10H. When the photomultiplier tube 42H receives the first detection light L1 at any regional position of the detection stage 10H, the photomultiplier tube 42H will generate a regional positioning signal, which includes the regional position and fluorescence signal intensity of the at least one target biological particle on the detection stage 10H, and is transmitted to the processor until all regional positions are scanned.

[0164] Step E: The processor determines that at least one of the regional positions on the detection carrier 10H has at least one target biological particle based on at least one of the regional positioning signals, and transmits at least one of the regional positioning signals to the charge coupled device 44H.

[0165] Step F: The charge-coupled device 44 moves relative to the detection platform 10 according to at least one of the regional positioning signals, and receives the second detection light L2 at each of the regional positions to sense the image position of at least one target biological particle on the detection platform 10, thereby generating an image signal respectively.

[0166] Step G: The processor obtains the precise position of the at least one target biological particle in at least one of the regional positions on the detection stage 10H according to the image signal.

[0167] The charge coupled device 44H of the detection device 100H of the ninth embodiment of the present invention is independent of the photomultiplier tube 42H, so it can achieve a faster detection effect of the multiple biological particles on the carrier 10H compared to the existing charge coupled device detection device, thereby accelerating the operation speed of the entire device.

[0168] Please refer to Fig.13As shown, the detection device 100I of the tenth preferred embodiment of the present invention is different from the detection device 100I of the tenth preferred embodiment and the detection device 100H of the ninth embodiment in that the filter spectrometer group of the detection device 100I of the tenth preferred embodiment further includes another objective lens 32I, and the objective lens 32I is arranged above the detection carrier 10I and below the color separation mirror 34I. In this way, when the photomultiplier tube 42I scans and senses the regional position of the at least one target biological particle and receives the first detection light L1, the regional range obtained by the photomultiplier tube 42I is smaller, so that the scanning accuracy is improved.

[0169] In addition, in order to avoid the height of the whole device being too high, the excitation light source 20I in this embodiment is arranged on one side of the dichroic mirror 34I, so that the difference between the detection device 100I of the tenth preferred embodiment and the detection device 100H of the ninth embodiment is that: in the detection device 100H of the ninth embodiment, the excitation light La of the excitation light source 20H penetrates the dichroic mirror 34H to the detection stage 10H, and the dichroic mirror 34H is located on the optical path of the first detection light L1 and is used to reflect the first detection light L1 into the first filter group 36H. In the detection device 100I of the tenth preferred embodiment, the excitation light La of the excitation light source 20I is reflected by the dichroic mirror 34I, and the excitation light La is focused by the objective lens 32I to irradiate the multiple biological particles on the detection stage 10I, and the dichroic mirror 34I is located on the optical path of the first detection light L1, and the first detection light L1 penetrates the objective lens 32I and the dichroic mirror 34I and enters the first filter group 36I.

[0170] The positions of the photomultiplier tube 42I and the first filter group 36I for receiving the first detection light L1 are the same as those of the components in the ninth embodiment, and the positions of the objective lens 32I, the second filter group 37I and the charge-coupled device 44I for receiving the second detection light L2 located below the detection platform 10I are also the same as those of the components in the ninth embodiment, so they are not repeated here.

[0171] Please refer to Fig.14As shown, the detection device 100J of the eleventh preferred embodiment of the present invention is different from the detection device 100I of the tenth preferred embodiment in that no objective lens is arranged between the detection stage 10J and the dichroic mirror 34J of the detection device 100J. Thus, compared with the mechanism of the detection device 100I of the tenth preferred embodiment, the overall volume is smaller and the weight is lighter. In the detection device 100J of the eleventh preferred embodiment, the excitation light La of the excitation light source 20J is reflected to the detection stage 10J by the dichroic mirror 34J, and the dichroic mirror 34J is located on the optical path of the first detection light L1, and the first detection light L1 enters the first filter group 36J after penetrating the dichroic mirror 34J, and the first detection light L1 is then received by the photomultiplier tube 42J. The remaining components of the detection device 100J of the eleventh preferred embodiment are the same as those of the detection device 100I of the tenth preferred embodiment, and are not described in detail.

[0172] In summary, through the above structural design, the detection device only needs one excitation light source and a set of filter spectroscopes to achieve the purpose of simultaneously locating the precise position of at least one target biological particle by scanning the photomultiplier tube and charge-coupled device of the sensor group. Compared with the existing mechanism design that each sensor must be equipped with a set of independent excitation light sources and filter spectroscopes, the overall volume of the detection device of the present invention is smaller, the weight is reduced, and the cost is relatively low. And the charge-coupled device and the photomultiplier tube are integrated into a mechanical structure of a scanning sensor group to make its movement during scanning and detection more accurate, and will not produce interference in mechanical movement. In addition, the photomultiplier tube in the present invention first quickly scans to determine whether there is at least one target biological particle, and locates the preliminary position of at least one target biological particle, and the charge-coupled device then finely locates the precise position of at least one target biological particle, thereby achieving the effect of high-efficiency detection and high detection accuracy.

[0173] The above description is only a preferred feasible embodiment of the present invention. Any equivalent changes made by applying the present invention specification and the scope of patent application should be included in the patent scope of the present invention.

[0174] Description of Reference Numerals

[0175] [First embodiment of the present invention]

[0176] 100: Detection equipment 10: Detection stage 20: Excitation light source

[0177] 30: Filter beam splitter group 32: Objective lens 34: Color dichroic mirror

[0178] 36: Filter set 38: Spectrometer 40: Scanning sensor set

[0179] 42: Photomultiplier tube 44: Charge coupled device L1: First detection light

[0180] L2: Second detection light La: Excitation light Lb: Radiation light

[0181] S1: First light path S2: Second light path

[0182] [Second embodiment of the present invention]

[0183] 100A: Detection equipment 10A: Detection stage 20A: Excitation light source

[0184] 30A: Filter beam splitter group 32A: Objective lens 34A: Color dichroic mirror

[0185] 36A: First filter group 37A: Second filter group

[0186] 38A: Spectroscope 42A: Photomultiplier tube 44A: Charge coupled device

[0187] [Third Embodiment of the Invention]

[0188] 100B: Detection equipment 10B: Detection stage 20B: Excitation light source

[0189] 32B: Objective lens 34B: Color separation mirror 36B: Filter set

[0190] 38B: Spectroscope 42B: Photomultiplier tube 44B: Charge coupled device

[0191] [Fourth embodiment of the present invention]

[0192] 100C: Detection equipment 10C: Detection stage 20C: Excitation light source

[0193] 32C: Objective lens 34C: Dichroic mirror 36C: First filter group

[0194] 37C: Second filter set 38C: Spectroscope 42C: Photomultiplier tube

[0195] 44C: Charge-coupled device

[0196] [Fifth Embodiment of the Invention]

[0197] 100D: Detection equipment 10D: Detection stage 20D: Excitation light source

[0198] 30D: filter spectroscope group 32D: objective lens 36D: filter group

[0199] 38D: Spectroscope 42D: Photomultiplier tube 44D: Charge coupled device

[0200] [Sixth embodiment of the present invention]

[0201] 100E: Detection equipment 10E: Detection stage 20E: Excitation light source

[0202] 32E: Objective lens 34E: Color dichroic mirror 36E: Filter set

[0203] 38E: Spectroscope 42E: Photomultiplier tube 44E: Charge coupled device

[0204] L11: First detection light band

[0205] [Seventh embodiment of the present invention]

[0206] 100F: Detection equipment 10F: Detection stage 20F: Excitation light source

[0207] 32F: Objective lens 34F: Color dichroic mirror 36F: Filter set

[0208] 38F: beam splitter 42F: photomultiplier tube 44F: charge coupled device

[0209] L11: First detection light band

[0210] [Eighth Embodiment of the Present Invention]

[0211] 100G: Detection equipment 10G: Detection platform 20G: Excitation light source

[0212] 32G: Objective lens 34G: Color splitter 38G: Beam splitter

[0213] 42G: Photomultiplier tube 44G: Charge coupled device L11: First detection light band

[0214] L21: Second detection light band

[0215] [Ninth Embodiment of the Invention]

[0216] 100H: Detection equipment 10H: Detection stage 20H: Excitation light source

[0217] 32H: Objective lens 34H: Color separation mirror 36H: First filter group

[0218] 37H: Second filter group 42H: Photomultiplier tube 44H: Charge coupled device

[0219] [Tenth embodiment of the present invention]

[0220] 100I: Detection equipment 10I: Detection stage 20I: Excitation light source

[0221] 32I: Objective lens 34I: Color separation mirror 36I: First filter group

[0222] 37I: Second filter group 42I: Photomultiplier tube 44I: Charge coupled device

[0223] [Eleventh Embodiment of the Present Invention]

[0224] 100J: Detection equipment 10J: Detection stage 20J: Excitation light source

[0225] 34J: Color dichroic mirror 36J: First filter group 42J: Photomultiplier tube

Claims

1. A detection device for detecting biological particles, comprising: a detection carrier for placing a plurality of biological particles; wherein the plurality of biological particles include at least one target biological particle; and An optical system comprising: an excitation light source, for providing an excitation light to illuminate the plurality of biological particles, and the at least one target biological particle among the plurality of biological particles is illuminated by the excitation light and excited to generate a radiation light; a filter spectroscope assembly, comprising a spectroscope, wherein the spectroscope is located on the optical path of the radiation light and is used to split the incident radiation light into a first detection light and a second detection light that account for different proportions of the radiation light, wherein the ratio of the first detection light to the radiation light is between 0.1 and 0.5, and the ratio of the second detection light to the radiation light is between 0.5 and 0.9; at least one photomultiplier tube is used to receive the first detection light; when receiving the first detection light, the at least one photomultiplier tube generates a regional positioning signal, wherein the regional positioning signal includes the regional position of the at least one target biological particle on the detection carrier; and At least one charge-coupled device is used to receive the second detection light. When receiving the second detection light, the at least one charge-coupled device generates an image signal, and the image signal includes the image position of the at least one target biological particle on the detection carrier.

2. The detection device for detecting biological particles according to claim 1, in, It includes a processor, which is electrically connected to the optical system and is used to receive the regional positioning signal and the image signal; when the processor receives the regional positioning signal, the processor transmits the regional positioning signal to the at least one charge-coupled device, so that the at least one charge-coupled device moves relative to the detection carrier according to the regional positioning signal, and the charge-coupled device generates the image signal.

3. The detection device for detecting biological particles according to claim 1, in, The filter spectroscope assembly includes an objective lens, which is located on the optical path of the radiated light and between the detection stage and the at least one charge coupled device.

4. The detection device for detecting biological particles according to claim 1, in, The detection platform is located on the light path of the excitation light, so that the excitation light directly irradiates the plurality of biological particles.

5. The detection device for detecting biological particles according to claim 1, in, The filter spectroscope group includes at least one color splitter, which is located on the light path of the excitation light and is used to reflect the excitation light so that the excitation light irradiates the plurality of biological particles after being reflected.

6. The detection device for detecting biological particles according to claim 5, in, The number of the at least one color separation mirror is multiple, the number of the at least one photomultiplier tube is multiple, and the number of the at least one type of target biological particles is multiple; wherein at least one of the multiple color separation mirrors is located on the light path of the first detection light, and is used to separate the incident first detection light into multiple first detection light bands of different bands for emission, and the multiple first detection light bands are respectively received by at least two of the multiple photomultiplier tubes, thereby the multiple photomultiplier tubes can respectively generate a regional positioning signal, and the multiple regional positioning signals respectively include the regional positions of the multiple target biological particles on the detection carrier.

7. The detection device for detecting biological particles according to claim 5, in, The number of the at least one color separation mirror is multiple, the number of the at least one charge-coupled device is multiple, and the number of types of the at least one target biological particle is multiple; wherein at least one of the multiple color separation mirrors is located on the light path of the second detection light, and is used to divide the incident second detection light into multiple second detection light bands of different bands for emission, and the multiple second detection light bands are respectively received by at least two of the multiple charge-coupled devices, thereby the multiple charge-coupled devices can respectively generate an image signal, and the multiple image signals include the image position of at least one of the multiple target biological particles on the detection carrier.

8. The detection device for detecting biological particles according to claim 1, in, The filter spectroscope group includes at least one filter group, and the at least one filter group is located on the optical path of the radiated light, the optical path of the first detection light, the optical path of the second detection light, or the optical path of a combination thereof.

9. A detection method for a detection device, used for a detection device as claimed in claim 1; the detection device further comprises a processor electrically connected to the optical system; in, The detection method comprises the following steps: Step A: the excitation light source emits the excitation light to illuminate the plurality of biological particles; Step B: the at least one target biological particle among the plurality of biological particles absorbs the excitation light to generate the radiated light, and the radiated light is incident on the filter spectroscope group; Step C: the filter spectroscope group separates the radiated light into the first detection light and the second detection light, and the first detection light and the second detection light are incident on the at least one photomultiplier tube and the at least one charge coupled device respectively; Step D: the at least one photomultiplier tube receives the first detection light and generates a regional positioning signal; Step E: the processor determines that at least one area position on the detection stage has at least one target biological particle according to the area positioning signal; Step F: the at least one charge coupled device receives the second detection light and generates an image signal; Step G: The processor obtains the precise position of the at least one target biological particle in the at least one regional position on the detection stage according to the image signal.

10. The detection method of the detection device according to claim 9, in, Step D includes the following steps: The at least one photomultiplier tube moves relative to the detection platform to scan multiple regional positions on the detection platform; when the photomultiplier tube receives the first detection light at at least one regional position of the detection platform, the photomultiplier tube generates at least one regional positioning signal.

11. The detection method of the detection device according to claim 10, in, Step E includes the following steps: The processor transmits at least one of the region positioning signals to the at least one charge coupled device.

12. The detection method of the detection device according to claim 11, in, Step F includes the following steps: The at least one charge-coupled device moves relative to the detection platform according to at least one of the regional positioning signals, and receives the second detection light to sense the image position of the at least one target biological particle on the detection platform, thereby generating at least one of the image signals.

13. The detection method of the detection device according to claim 11, in, After the at least one photomultiplier tube in step D scans all the regional positions on the detection carrier, step E will be executed continuously. The processor in step E determines which regional positions on the detection carrier generate the regional positioning signal based on at least one of the regional positioning signals, and then step F will be executed continuously. The at least one charge-coupled device in step F moves relative to the detection carrier to sequentially generate an image signal of at least one regional position on the detection carrier having at least one target biological particle.

14. The detection method of the detection device according to claim 9, in, The optical system's filter spectroscope assembly includes the spectroscope and an objective lens, wherein the spectroscope separates the radiated light into the first detection light and the second detection light, and the objective lens is located on the optical path of the radiated light and between the detection stage and the at least one charge-coupled device; the detection method includes the following steps between step B and step C: The objective lens causes the emitted light to enter the beam splitter.

15. The detection method of the detection device according to claim 9, in, The optical system's filter spectroscope assembly includes a color splitter, and the color splitter is located on the light path of the excitation light. The detection method includes the following steps in step A: The dichroic mirror reflects the excitation light and then irradiates the plurality of biological particles.

16. The detection method of the detection device according to claim 9, in, The optical system's filter spectroscope group includes a filter group and the spectroscope, the filter group and the spectroscope are located on the light path of the radiated light, wherein the spectroscope splits the radiated light into the first detection light and the second detection light for emission; the detection method includes the following steps between step B and step C: The filter set can only allow the wavelength band of the radiated light to pass through, so that the radiated light can be incident on the spectroscope after passing through.

17. The detection method of the detection device according to claim 9, in, The optical system's filter spectroscope group includes a first filter group, a second filter group, and the spectroscope, wherein the first filter group and the second filter group are respectively located on the optical paths of the first detection light and the second detection light, and the spectroscope is located on the optical path of the radiated light, wherein the spectroscope splits the radiated light into the first detection light and the second detection light for emission; the detection method includes the following steps between step C and step D: The first filter group can only allow the wavelength band of the first detection light to pass through, so that the first detection light is incident on the at least one photomultiplier tube after passing through; The second filter group can only allow the wavelength band of the second detection light to pass through, so that the second detection light is incident on the at least one charge coupled device after passing through.

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