High-throughput wide-area Raman spectrum system for bacterial colony detection and implementation method

Abstract

The invention provides a high-throughput wide-area Raman spectrum system for bacterial colony detection and an implementation method. The high-throughput wide-area Raman spectrum system comprises a laser and light spot adjusting module, a Raman imaging module and a detection module, the spatial heterogeneity problem of a bacterial colony sample is solved by setting a light spot adjustment scheme, a bacterial colony detection scheme and a high-throughput wide-area Raman scheme, so that the spectral repeatability of the bacterial colony sample is better, and establishment of a spectral database and identification of the bacterial colony sample are facilitated; the system is different from a common wide-area Raman system, has high throughput and high resolution, is more suitable for analysis and identification of biological samples, meets the resolution requirements of analysis and identification of various biological samples, and is scientific and reasonable in overall structural design, safe and reliable in implementation process and suitable for popularization and application.

Description

Technical Field

[0001] The invention belongs to the technical field of spectral analysis, and in particular to a high-throughput wide-area Raman spectroscopy system for colony detection and an implementation method thereof. Background Art

[0002] At present, most detection methods for colonies are based on single-point detection of confocal Raman spectroscopy systems. The diameter of the focused light spot under the objective lens of a traditional confocal Raman spectroscopy system is about 1-2μm, while the diameter of the colony is often between 0.2-2mm. Studies have shown that there is heterogeneity in different positions of the colony, and the spectrum collected at a single point is difficult to characterize the information of the entire colony.

[0003] Therefore, under the existing detection methods, the detection results of Raman spectra of the same colony are often different; the spatial heterogeneity of different positions of the colonies has greatly hindered the repeatability and stability of the colony Raman spectra, which has become a technical problem that needs to be solved urgently. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a high-throughput wide-area Raman spectroscopy system for colony detection and an implementation method thereof. The system can not only adjust the Raman signal collection range of the colony sample under the objective lens through the laser adjustment module, but also easily realize the confocality of the large light spot under the objective lens and the spectrometer entrance through the use of multi-core optical fiber, thereby ensuring that the Raman signals in the laser illumination area are fully received by the spectrometer; the circular-to-linear optical fiber structure can improve the system throughput while ensuring that the resolution of the Raman spectrum is not reduced, thereby ensuring the need for a certain resolution of the Raman spectrum of biological samples. The overall structural design of the system is scientific and reasonable, and has a broad scientific research application market.

[0005] A high-throughput wide-area Raman spectroscopy system and implementation method for bacterial colony detection, wherein:

[0006] A high-throughput wide-area Raman spectroscopy system for bacterial colony detection, comprising:

[0007] Laser and spot adjustment module, Raman imaging module, and detection module;

[0008] The laser and light spot adjustment module includes: a laser, a first collimating mirror, a first reflecting mirror, and a first focusing mirror;

[0009] The first collimating mirror is arranged on the upper side of the laser; the first reflecting mirror is arranged on the upper side of the first collimating mirror; the first focusing mirror is arranged on the right side of the first reflecting mirror;

[0010] As an example, the laser used is a diode laser with a wavelength of 785 nm, which can effectively suppress the high fluorescence of the bacterial colony sample.

[0011] As an example, the first collimator is disposed on the upper side of the laser to control the divergence angle of the laser beam so that the laser is collimated or has a certain divergence angle.

[0012] The Raman imaging module includes: an imaging camera, an objective lens, an LED, a second collimating lens, a second focusing lens, a first beam splitter, a second beam splitter, a second reflecting mirror, a first filter, a second filter, a third focusing lens, and a third filter;

[0013] Wherein: the objective lens is directly above the sample; the second beam splitter is arranged above the objective lens; the first beam splitter is arranged on the right side of the second beam splitter; the left side of the first beam splitter is the second focusing lens, and the right side is the second collimating lens; the imaging camera is arranged on the lower side of the first beam splitter; the LED is arranged on the right side of the second collimating lens; the first filter and the second reflector are arranged above the second beam splitter in sequence; the second filter, the third focusing lens and the third filter are arranged on the right side of the second reflector in sequence;

[0014] As an example, the first beam splitter and the second beam splitter are both 50% beam splitters.

[0015] The detection module 103 includes: a circular-to-linear multi-core optical fiber, a spectrometer, and a spectral camera;

[0016] The circular end of the circular-to-linear multi-core optical fiber serves as a Raman signal receiving end and is located at the focus of the third focusing lens; the linear end of the circular-to-linear multi-core optical fiber is connected to the entrance of the spectrometer; the spectroscopic camera is arranged below the spectrometer; the NA of the spectrometer is 0.25, and the NA of the circular-to-linear multi-core optical fiber is 0.22. This arrangement can ensure that the Raman signal transmitted by the circular-to-linear multi-core optical fiber can be fully received by the spectroscopic camera.

[0017] As an example, the circular-to-linear transition means that: a fiber optic connector extends from one side of the entrance end of the multi-core optical fiber, and the multi-core optical fibers in the fiber optic connector are circular ends arranged in a circular manner; a fiber optic connector also extends from one side of the exit end, and the multi-core optical fibers in the fiber optic connector are linear ends arranged linearly.

[0018] As an example, the spectral camera is a CCD spectral camera.

[0019] As an example, the number of cores of the circular-to-linear multi-core optical fiber is 7 cores, specifically: one core is in the middle of the circular end, and the other six cores surround the middle core; one side of the outlet end has seven cores arranged vertically in a row; when the number of cores is 7, the core diameter of a single optical fiber is 105μm; the corresponding circular end face diameter is 380μm.

[0020] As an application example, the number of cores of the circular-to-linear multi-core optical fiber can also be: other core numbers greater than 1 core; as the number of optical fiber cores increases, the illumination range will be wider, and the number of optical fiber cores is limited by the target height of the spectral camera.

[0021] As an example, the wavelength range of the circular-to-linear multi-core optical fiber is 200-1000 nm.

[0022] A method for implementing high-throughput wide-area Raman spectroscopy for bacterial colony detection, comprising:

[0023] Step 1: Setting the light spot adjustment scheme;

[0024] When the first focusing mirror and / or the objective lens have different focal lengths, the light spot under the objective lens can be modulated to a certain size; when the first focusing mirror is removed from the optical path, the system is the same as the traditional confocal Raman.

[0025] Therefore, by replacing the first focusing mirror and / or the objective lens with different focal lengths to adjust the spot size, a method of quickly switching between the traditional confocal Raman system and the wide-area Raman spectroscopy system can be achieved;

[0026] As an example, the divergence angle of the light beam can also be controlled by controlling the distance between the laser and the first collimator, so that the light spot under the objective lens can be continuously adjusted. When the traditional confocal system is to be restored, the distance between the laser and the first collimator is initially reset.

[0027] Step 2: Colony detection setting;

[0028] The laser emitted by the laser passes through the first collimator, the first reflector, the first focusing mirror, and the first filter, and then acts on the sample through the objective lens to stimulate Raman light of the sample, that is, Raman signal;

[0029] The Raman signal return optical path passes through the first filter, the second reflector, and the second filter to filter out stray light;

[0030] Step 3: High-throughput wide-area Raman solution setup;

[0031] Then, it is focused by a third focusing lens and connected to the circular end of the circular-to-linear multi-core optical fiber; the third optical filter is located on the left side of the circular end of the circular-to-linear multi-core optical fiber, and is used to filter out stray light from the external environment;

[0032] After the Raman signal enters the spectrometer from the linear end of the circular-to-linear multi-core optical fiber, the Raman signal is separated and expanded into a spectral signal, which is then received by a spectral camera, thereby finally realizing the collection of high-throughput wide-area Raman signals;

[0033] In order to achieve the conjugation of the laser illumination area with the circular end of the circular-to-linear multi-core optical fiber, the diameter D of the laser illumination areal The circular end diameter D of the circular-to-linear multi-core optical fiber f There is a corresponding relationship.

[0034] As an example, the corresponding relationship is determined according to the focal length f1 of the objective lens and the focal length f2 of the third focusing lens, and the calculation formula is as follows:

[0035]

[0036] Thus, according to the circular end diameter of the circular-to-linear multi-core optical fiber, the size of the required laser illumination area can be calculated according to Formula 1.

[0037] As an example of application, when the focal length f2 of the third focusing lens is 19 mm, the diameter D of the circular end of the circular-to-linear multi-core optical fiber is f =380μm, then:

[0038] When the focal length of the objective lens is f1 = 3.6 mm, the required diameter of the laser illumination area is D l =72μm;

[0039] When the focal length of the objective lens is f1 = 9 mm, the required diameter D of the laser illumination area is l =180μm;

[0040] As an example, when the number of cores of the circular-to-linear multi-core optical fiber is 7, the corresponding laser illumination area can be calculated based on the signal receiving area of ​​the 7-core optical fiber. As the number of optical fiber cores increases, the sample detection range will increase accordingly.

[0041] As an example, the laser illumination area is the sample detection range.

[0042] Beneficial effects of the present invention:

[0043] The wide-area Raman system of the present invention is different from the conventional confocal Raman system. The laser illumination area can be flexibly adjusted through the laser adjustment module to solve the spatial heterogeneity problem of the colony sample, thereby making the spectrum repeatability of the colony sample better and facilitating the establishment of the spectrum database and the identification of the colony sample.

[0044] The present invention is different from the common wide-area Raman system, taking into account both high throughput and high resolution, and is more suitable for the analysis and identification of biological samples. By using multi-core optical fiber to couple the Raman signal into the spectrometer module, a simple confocusing of the light spot under the mirror and the spectrometer entrance is achieved, that is, the full reception of the Raman signal in the laser illumination area is achieved. At the same time, the circular-to-linear optical fiber structure can ensure that the resolution of the Raman spectrum is less than 9.7 cm -1 , meeting the resolution requirements for analysis and identification of biological samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 The overall structural design diagram of a high-throughput wide-area Raman spectroscopy system for colony detection of the present invention.

[0046] Figure 2 This is a reference diagram of measured data results of Example 1 of a high-throughput wide-area Raman spectroscopy implementation method for colony detection of the present invention.

[0047] Figure 3 The present invention is a statistical diagram of the spot diameter data under the objective lens when the first focusing lens and the objective lens with different focal lengths are replaced in the spot adjustment scheme setting of the high-throughput wide-area Raman spectroscopy system for colony detection of the present invention.

[0048] Figure 4 This is a test data diagram comparing the Raman signal intensity of a next-order silicon peak with different spot sizes in a high-throughput wide-area Raman spectroscopy system for colony detection of the present invention.

[0049] Figure 5 This is a comparison diagram of the average Raman spectra of the measured results of 6 bacterial colony samples in Example 2 of a high-throughput wide-area Raman spectroscopy implementation method for colony detection of the present invention. DETAILED DESCRIPTION

[0050] Below, reference Figures 1 to 5 As shown, a high-throughput wide-area Raman spectroscopy system and implementation method for colony detection, wherein:

[0051] A high-throughput wide-area Raman spectroscopy system for bacterial colony detection, referring to Figure 1 As shown, it includes: a laser and light spot adjustment module 101, a Raman imaging module 102, and a detection module 103;

[0052] The laser and light spot adjustment module 101 includes: a laser 104, a first collimating mirror 105, a first reflecting mirror 106, and a first focusing mirror 107;

[0053] The first collimating mirror 105 is disposed on the upper side of the laser 104; the first reflecting mirror 106 is disposed on the upper side of the first collimating mirror 105; the first focusing mirror 107 is disposed on the right side of the first reflecting mirror 106;

[0054] As an example, the laser used is a diode laser with a wavelength of 785 nm, which can effectively suppress the high fluorescence of the bacterial colony sample.

[0055] As an example, the first collimator 105 is disposed on the upper side of the laser 104 to control the divergence angle of the laser beam so that the laser is collimated or has a certain divergence angle.

[0056] The Raman imaging module 102 includes: an imaging camera 108, an objective lens 109, an LED 110, a second collimating lens 111, a second focusing lens 112, a first beam splitter 113, a second beam splitter 114, a second reflecting mirror 115, a first filter 116, a second filter 117, a third focusing lens 118, and a third filter 119;

[0057] Wherein: the objective lens 109 is directly above the sample; the second beam splitter 114 is arranged above the objective lens 109; the first beam splitter 113 is arranged on the right side of the second beam splitter 114; the left side of the first beam splitter 113 is the second focusing mirror 112, and the right side is the second collimating mirror 111; the imaging camera 108 is arranged on the lower side of the first beam splitter 113; the LED 110 is arranged on the right side of the second collimating mirror 111; the first filter 116 and the second reflector 115 are arranged above the second beam splitter 114 in sequence; the right side of the second reflector 115 is arranged on the second filter 117, the third focusing mirror 118 and the third filter 119 in sequence;

[0058] As an example, the first beam splitter 113 and the second beam splitter 114 are both 50% beam splitters.

[0059] The detection module 103 includes: a circular-to-linear multi-core optical fiber 120, a spectrometer 121, and a spectral camera 122;

[0060] The circular end of the circular-to-linear multi-core optical fiber 120 serves as a Raman signal receiving end and is located at the focus of the third focusing lens 118; the linear end of the circular-to-linear multi-core optical fiber 120 is connected to the entrance of the spectrometer 121; the spectral camera 122 is arranged below the spectrometer 121; the NA of the spectrometer 121 is 0.25, and the NA of the circular-to-linear multi-core optical fiber is 0.22. This arrangement can ensure that the Raman signal transmitted by the circular-to-linear multi-core optical fiber 120 can be fully received by the spectral camera 122.

[0061] As an example, the circular-to-linear transition means that: a fiber optic connector extends from one side of the entrance end of the multi-core optical fiber, and the multi-core optical fibers in the fiber optic connector are circular ends arranged in a circular manner; a fiber optic connector also extends from one side of the exit end, and the multi-core optical fibers in the fiber optic connector are linear ends arranged linearly.

[0062] As an example, the spectral camera is a CCD spectral camera.

[0063] As an example, the number of cores of the circular-to-linear multi-core optical fiber 120 is 7 cores, specifically: one core is in the middle of the circular end, and the other six cores surround the middle core; one side of the outlet end has seven cores arranged vertically in a row; when the number of cores is 7, the core diameter of a single optical fiber is 105μm; the corresponding circular end face diameter is 380μm.

[0064] As an application example, the number of cores of the circular-to-linear multi-core optical fiber 120 can also be: other core numbers greater than 1 core; as the number of optical fiber cores increases, the illumination range will be wider, and the number of optical fiber cores is limited by the target surface height of the spectral camera 122.

[0065] As an example, the wavelength range of the circular-to-linear multi-core optical fiber 120 is 200-1000 nm.

[0066] A method for implementing high-throughput wide-area Raman spectroscopy for bacterial colony detection, comprising:

[0067] Step 1: Set up the light spot adjustment scheme; see Figure 3 As shown;

[0068] When the first focusing mirror 107 and / or the objective lens 109 are selected with different focal lengths, the light spot under the objective lens can be modulated to a certain size; when the first focusing mirror 107 is removed from the optical path, the system is the same as the conventional confocal Raman.

[0069] Therefore, by replacing the first focusing mirror 107 and / or the objective lens 109 with different focal lengths to adjust the spot size, a fast switch between the conventional confocal Raman system and the wide-area Raman spectroscopy system can be achieved;

[0070] As an example, the divergence angle of the light beam can also be controlled by controlling the distance between the laser 104 and the first collimator 105, so that the continuous adjustment of the light spot under the objective lens can be achieved. When the traditional confocal system is to be restored, the distance between the laser 104 and the first collimator 105 is initially reset.

[0071] Step 2: Colony detection setting;

[0072] The laser light emitted by the laser passes through the first collimator 105, the first reflector 106, the first focusing lens 107, and the first filter 116, and then acts on the sample through the objective lens 109 to stimulate the Raman light of the sample, that is, the Raman signal;

[0073] The Raman signal return optical path passes through the first filter 116, the second reflector 115, and the second filter 117 to filter out stray light;

[0074] Step 3: High-throughput wide-area Raman solution setup;

[0075] Afterwards, it is focused by the third focusing lens 118 and connected to the circular end of the circular-to-linear multi-core optical fiber 120; the third optical filter 119 is located on the left side of the circular end of the circular-to-linear multi-core optical fiber 120, and is used to filter out stray light from the external environment;

[0076] After the Raman signal enters the spectrometer 121 from the linear end of the circular-to-linear multi-core optical fiber 120, the Raman signal is separated and expanded into a spectral signal, which is then received by the spectral camera 122, thereby finally realizing the collection of high-throughput wide-area Raman signals;

[0077] In order to achieve the conjugation of the laser illumination area with the circular end of the circular-to-linear multi-core optical fiber, the diameter D of the laser illumination area l The circular end diameter D of the circular-to-linear multi-core optical fiber 120 f There is a corresponding relationship.

[0078] As an example, the corresponding relationship is determined according to the focal length f1 of the objective lens 109 and the focal length f2 of the third focusing lens 118, and the calculation formula is as follows:

[0079]

[0080] Thus, according to the circular end diameter of the circular-to-linear multi-core optical fiber 120 , the size of the required laser illumination area can be calculated according to Formula 1.

[0081] As an example of application, when the focal length f2 of the third focusing lens 118 is 19 mm, the diameter D of the circular end of the circular-to-linear multi-core optical fiber 120 is f =380μm, then:

[0082] When the focal length f1 of the objective lens 109 is 3.6 mm, the diameter D of the required laser illumination area is l =72μm;

[0083] When the focal length f1 of the objective lens 109 is 9 mm, the diameter D of the required laser illumination area is l =180μm;

[0084] As an example, when the number of cores of the circular-to-linear multi-core optical fiber 120 is 7, the corresponding laser illumination area can be calculated based on the signal receiving area of ​​the 7-core optical fiber. As the number of optical fiber cores increases, the sample detection range will increase accordingly.

[0085] As an example, the laser illumination area is the sample detection range.

[0086] In order to better illustrate the advantages of the present invention, specific experiments are now used to illustrate the actual measured data results during operation.

[0087] Embodiment 1:

[0088] Reference Figure 2 As shown, the spectrum of the argon lamp is shown in Figure 1. The horizontal axis is the Raman shift after wave number conversion, showing that the half-peak width of the rightmost peak is 9.3 cm -1 The calculated average spectral resolution is about 9.7 cm -1 ;

[0089] The average spectral resolution is Figure 2 The average half-width of the eight peaks.

[0090] When the number of cores of the circular-to-linear multi-core optical fiber 120 is 7 and the core diameter of a single optical fiber is 105 μm, the collected Raman signal can achieve 9.7 cm -1 Spectral resolution;

[0091] This resolution result is much higher than the combination and splicing of existing technologies;

[0092] This resolution result is not only excellent, but also based on the structural design of adjustable spot, it can also efficiently measure any sample with different spot diameters;

[0093] Reference Figure 4 As shown, it can be seen that within a certain range of spot sizes, the intensity of the silicon peak does not change from small to large, which proves that the Raman signals excited within this range are fully received by the spectrometer 121 and the spectral camera 122, indicating that the circular end diameter of the circular-to-linear multi-core optical fiber 120 set according to Formula 1 is accurately calculated, and once again proves that according to Formula 1, the corresponding relationship can be easily adjusted to complete the excitation and collection of high-throughput wide-area Raman signals.

[0094] Embodiment 2:

[0095] Reference Figure 5 As shown, a high-throughput wide-area Raman spectroscopy implementation method for colony detection of the present invention is used to perform Raman analysis on 6 types of colony samples;

[0096] From the comparison of the average Raman spectra of the six bacterial colony samples, it can be seen that the average Raman spectra of different bacterial colonies are different, which can fully illustrate that the bacterial colony Raman spectra generated by the system of the present invention and the implementation method based on the system can be reliably used for accurate identification of bacterial colonies;

[0097] In addition, the high signal-to-noise ratio of the Raman spectra of the six bacterial colony samples further proves that the system of the present invention and the implementation method based on the system can produce high throughput and high sensitivity.

[0098] The wide-area Raman spectroscopy system proposed in the present invention is different from the conventional Raman spectroscopy system; the light spot under the microscope can be regulated by the original modification of the existing confocal Raman spectroscopy system; at the same time, the use of circular-to-linear multi-core optical fiber can realize simple confocalization of the large light spot under the microscope and the entrance of the spectrometer; without losing resolution, it is ensured that the Raman signals in the laser illumination area are all collected in the Raman spectrometer; this design can obtain a colony Raman spectrum with a high signal-to-noise ratio in a short time, and at the same time solves the spatial heterogeneity problem of the colony to a certain extent, which will be very beneficial to the Raman signal collection of colony samples.

[0099] The above are only preferred embodiments of the present invention. It should be understood that the description of the above embodiments is only used to help understand the method and core ideas of the present invention, and is not used to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, etc. made within the ideas and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A high-throughput wide-area Raman spectroscopy system for bacterial colony detection, characterized in that: include: Laser and spot adjustment module, Raman imaging module, and detection module; The laser and light spot adjustment module includes: a laser, a first collimating mirror, a first reflecting mirror, and a first focusing mirror; The first collimating mirror is arranged on the upper side of the laser; the first reflecting mirror is arranged on the upper side of the first collimating mirror; the first focusing mirror is arranged on the right side of the first reflecting mirror; The Raman imaging module includes: an imaging camera, an objective lens, an LED, a second collimating lens, a second focusing lens, a first beam splitter, a second beam splitter, a second reflecting mirror, a first filter, a second filter, a third focusing lens, and a third filter; Wherein: the objective lens is directly above the sample; the second beam splitter is arranged above the objective lens; the first beam splitter is arranged on the right side of the second beam splitter; the left side of the first beam splitter is the second focusing lens, and the right side is the second collimating lens; the imaging camera is arranged on the lower side of the first beam splitter; the LED is arranged on the right side of the second collimating lens; the first filter and the second reflector are arranged above the second beam splitter in sequence; the second filter, the third focusing lens and the third filter are arranged on the right side of the second reflector in sequence; The detection module includes: a circular-to-linear multi-core optical fiber, a spectrometer, and a spectral camera; The circular end of the circular-to-linear multi-core optical fiber serves as a Raman signal receiving end and is located at the focus of the third focusing lens; the linear end of the circular-to-linear multi-core optical fiber is connected to the entrance of the spectrometer; the spectroscopic camera is arranged below the spectrometer; the NA of the spectrometer is 0.25, and the NA of the circular-to-linear multi-core optical fiber is 0.

22. This arrangement can ensure that the Raman signal transmitted by the circular-to-linear multi-core optical fiber can be fully received by the spectroscopic camera.

2. A high-throughput wide-area Raman spectroscopy system for colony detection according to claim 1, characterized in that: The laser used is a diode laser, and the wavelength of the diode laser is 785nm.

3. A high-throughput wide-area Raman spectroscopy system for colony detection according to claim 1, characterized in that: The first beam splitter and the second beam splitter are both 50% beam splitters.

4. A high-throughput wide-area Raman spectroscopy system for colony detection according to claim 1, characterized in that: The circular-to-linear transition means that: a fiber optic connector extends from one side of the entrance end of the multi-core optical fiber, and the multi-core optical fibers in the fiber optic connector are circular ends arranged in a circular manner; a fiber optic connector also extends from one side of the exit end, and the multi-core optical fibers in the fiber optic connector are linear ends arranged in a linear manner.

5. A high-throughput wide-area Raman spectroscopy system for colony detection according to claim 1, characterized in that: The spectral camera is a CCD spectral camera.

6. A high-throughput wide-area Raman spectroscopy system for colony detection according to claim 1, characterized in that: The number of cores of the circular-to-linear multi-core optical fiber 120 is 7 cores, specifically: one core is in the middle of the circular end, and the other six cores surround the middle core; one side of the outlet end has seven cores arranged vertically in a row; when the number of cores is 7, the core diameter of a single optical fiber is 105μm; the corresponding circular end face diameter is 380μm.

7. A high-throughput wide-area Raman spectroscopy system for colony detection according to claim 1, characterized in that: The wavelength range of the circular-to-linear multi-core optical fiber is 200-1000nm.

8. A method for implementing high-throughput wide-area Raman spectroscopy for colony detection according to any one of claims 1 to 7, characterized in that: include: Step 1: Setting the light spot adjustment scheme; When the first focusing mirror and / or the objective lens have different focal lengths, the light spot under the objective lens can be modulated to a certain size; when the first focusing mirror is removed from the optical path, the system is the same as the traditional confocal Raman. Therefore, by replacing the first focusing mirror and / or the objective lens with different focal lengths to adjust the spot size, a method of quickly switching between the traditional confocal Raman system and the wide-area Raman spectroscopy system can be achieved; Step 2: Colony detection setting; The laser emitted by the laser passes through the first collimator, the first reflector, the first focusing mirror, and the first filter, and then acts on the sample through the objective lens to stimulate Raman light of the sample, that is, Raman signal; The Raman signal return optical path passes through the first filter, the second reflector, and the second filter to filter out stray light; Step 3: High-throughput wide-area Raman solution setup; Then, it is focused by a third focusing lens and connected to the circular end of the circular-to-linear multi-core optical fiber; the third optical filter is located on the left side of the circular end of the circular-to-linear multi-core optical fiber, and is used to filter out stray light from the external environment; After the Raman signal enters the spectrometer from the linear end of the circular-to-linear multi-core optical fiber, the Raman signal is separated and expanded into a spectral signal, which is then received by a spectral camera, thereby finally realizing the collection of high-throughput wide-area Raman signals; In order to achieve the conjugation of the laser illumination area with the circular end of the circular-to-linear multi-core optical fiber, the diameter D of the laser illumination area l The circular end diameter D of the circular-to-linear multi-core optical fiber f There is a corresponding relationship.

9. A method for realizing high-throughput wide-area Raman spectroscopy for colony detection according to claim 8, characterized in that: The divergence angle of the light beam is controlled by controlling the distance between the laser and the first collimator, thereby achieving continuous adjustment of the light spot under the objective lens. When the traditional confocal system is to be restored, the distance between the laser and the first collimator is initially reset.

10. The method for realizing high-throughput wide-area Raman spectroscopy for colony detection according to claim 8, characterized in that: The corresponding relationship is determined according to the focal length f1 of the objective lens and the focal length f2 of the third focusing lens, and the calculation formula is as follows: Thus, according to the circular end diameter of the circular-to-linear multi-core optical fiber, the size of the required laser illumination area can be calculated according to Formula 1.

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