Optical measuring device

Abstract

The invention discloses an optical measuring device. A sheath flow assembly of the optical measuring device is used for testing a to-be-tested sample flow; the laser assembly comprises at least two lasers with different wavelength ranges and is used for emitting laser to the sheath flow assembly, so that the laser irradiates a to-be-detected sample flow and excites the to-be-detected sample flow to generate a plurality of fluorescent light beams with different wavelengths; the fluorescence detection assembly is used for receiving fluorescence beams; the first shaping assembly is located between the fluorescent light receiving assembly and the sheath flow assembly and used for homogenizing and shaping the fluorescent light beams. According to the mode, the first shaping assembly is arranged between the fluorescence receiving assembly and the sheath flow assembly, and the first shaping assembly is used for homogenizing and shaping the fluorescence light beams excited by the sheath flow assembly, so that the chromatic aberration between the fluorescence light beams with different wavelengths excited by at least two lasers with different wavelength ranges is reduced; the signal-to-noise ratio of the fluorescence receiving assembly is improved, and the accuracy of optical measurement is further improved.

Description

Technical Field

[0001] This application relates to the field of optical measurement technologies, and particularly to an optical measurement device. Background Art

[0002] Existing optical measurement devices can classify and quantitatively analyze and detect cell apoptosis, drug-resistant proteins, oncogene protein products, cell surface markers, cell proliferation markers, etc. through flow cytometry, and are widely used with great development prospects.

[0003] However, the existing optical measurement devices have limited numbers of optical paths for fluorescence detection. When using laser beams with multiple different wavelength ranges for fluorescence detection, the laser beams with different wavelength ranges cannot be accurately irradiated to designated positions, and the detection accuracy of the optical measurement device is low. Summary of the Invention

[0004] To solve the problem of low detection accuracy of the optical measurement device in the existing technology, this application provides an optical measurement device.

[0005] To solve the above problems, this application provides an optical measurement device, including a sheath flow assembly, a laser assembly, a fluorescence receiving assembly, and a first shaping assembly. The above sheath flow assembly is used to test a sample flow to be measured; the above laser assembly is disposed on one side of the above sheath flow assembly and includes at least two lasers with different wavelength ranges, and is used to emit laser to the above sheath flow assembly, so that the laser irradiates the above sample flow to be measured and excites the above sample flow to be measured to generate multiple fluorescence beams with different wavelengths; the above fluorescence receiving assembly is disposed on the side of the above sheath flow assembly away from the above laser assembly, and the above fluorescence detection assembly is used to receive the above fluorescence beams; the above first shaping assembly is located between the above fluorescence receiving assembly and the above sheath flow assembly, and is used to homogenize and shape the above fluorescence beams to reduce the chromatic aberration between the above fluorescence beams with different wavelengths excited by at least two lasers with different wavelength ranges.

[0006] Optionally, the above first shaping assembly includes at least one microlens array and a collecting lens, the above fluorescence receiving assembly includes a first beam splitter, the above fluorescence beams excited by the above sheath flow assembly sequentially pass through the above microlens array and the above collecting lens and enter the above first beam splitter, the above microlens array homogenizes and shapes the above fluorescence beams through a plurality of microlens sub-units, the above collecting lens is used to collect the shaped above fluorescence beams, and the above first beam splitter is used to split the above fluorescence beams with different wavelengths.

[0007] Optionally, the distance between the above microlens array and the light-emitting surface of the above sheath flow assembly is 4 - 8 mm, and the diameter of the above microlens sub-units is 2.5 - 2.8 mm.

[0008] Optionally, the above optical measurement device further includes a collection component, which is disposed between the first shaping component and the sheath flow component. The collection component is used to focus the fluorescence beam excited by the sheath flow component and transmit the focused fluorescence beam to the first shaping component. Wherein, the distance between the microlens array and the light-emitting surface of the collection component is 4-8 mm, and the diameter of the microlens sub-unit is 2.5-2.8 mm.

[0009] Optionally, the laser beam emitted by the laser component is incident on the sheath flow component along a first direction and excites a sample flow to be measured to generate a fluorescence beam and a scattered beam. The fluorescence receiving component includes a first filter, a first receiver, a second filter, and a second receiver. The first receiver is used to receive the fluorescence beam filtered by the first filter, and the second receiver is used to receive the fluorescence beam filtered by the second filter. The optical measurement device further includes a third filter and a scattered light receiver, and the scattered light receiver is used to receive the scattered beam filtered by the third filter. Wherein, at least one of the first filter, the second filter, and the third filter has a first distance from the adjacent receiver of the filter close to the sheath flow component in the first direction, and / or the scattered light receiver has a second distance from the first receiver and the second receiver respectively in a second direction, and the second direction is the flowing direction of the sample flow to be measured in the sheath flow component.

[0010] Optionally, the distance between the scattered light receiver and the first filter in the first direction is the first distance; and / or the distance between the first receiver and the second filter in the first direction is the first distance, and the first distance is 30-80 mm.

[0011] Optionally, the wavelength range of the first fluorescence beam received by the first receiver is smaller than the wavelength range of the second fluorescence beam received by the second receiver, and / or the scattered light receiver is used to receive the scattered beam and obtain the forward scattered light intensity of the scattered beam.

[0012] Optionally, the fluorescence receiving component is disposed on one side of the sheath flow component along the second direction. The fluorescence beam includes a first fluorescence beam and a second fluorescence beam with different wavelengths. The first filter is used to filter the first fluorescence beam so that the filtered first fluorescence beam is incident on the first receiver along the first direction, and the second filter is used to filter the second fluorescence beam so that the filtered second fluorescence beam is incident on the second receiver along the first direction. Wherein, the laser component, the sheath flow component, the third filter, and the scattered light receiver are sequentially disposed along the first direction, the projection positions of the third filter, the first filter, and the second filter are arranged in sequence in the first direction, and the scattered light receiver, the first receiver, and the second receiver are spaced apart in the second direction.

[0013] Optionally, the second distance is 10-30 mm.

[0014] Optionally, the above-mentioned fluorescence receiving component includes a plurality of first beam splitters, and the plurality of first beam splitters are used to separate the above-mentioned fluorescence beam at different wavelengths. The plurality of first beam splitters are arranged in sequence along the optical path of the above-mentioned fluorescence beam, so that the above-mentioned fluorescence beams of different wavelengths are used for classification testing and / or quantitative testing of the above-mentioned sample stream to be measured.

[0015] The present application provides an optical measurement device. The sheath flow component of the optical measurement device is used to test the sample stream to be measured; the laser component includes at least two lasers with different wavelength ranges, which are used to emit laser to the sheath flow component, so that the laser irradiates the sample stream to be measured and excites the sample stream to be measured to generate a plurality of fluorescence beams with different wavelengths; the fluorescence detection component is used to receive the fluorescence beams; the first shaping component is located between the fluorescence receiving component and the sheath flow component and is used to homogenize and shape the fluorescence beams. In the above manner, the first shaping component is arranged between the fluorescence receiving component and the sheath flow component, and the fluorescence beams excited by the sheath flow component are homogenized and shaped by the first shaping component, so as to reduce the chromatic aberration between the fluorescence beams of different wavelengths excited by at least two lasers with different wavelength ranges, improve the signal-to-noise ratio of the fluorescence receiving component, and further improve the accuracy of optical measurement. Description of the Drawings

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present application. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings. Among them:

[0017] Figure 1 is a schematic structural diagram of the first embodiment of the optical measurement device provided by the present application;

[0018] Figure 2 is Figure 1 a schematic structural diagram of an embodiment of the first shaping component in

[0019] Figure 3 is a schematic structural diagram of the second embodiment of the optical measurement device provided by the present application;

[0020] Figure 4 is a schematic structural diagram of the third embodiment of the optical measurement device provided by the present application;

[0021] Figure 5 is a schematic structural diagram of the fourth embodiment of the optical measurement device provided by the present application. Detailed Embodiments

[0022] Next, the technical solutions in the embodiments of the present application will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present application.

[0023] It should be noted that if there are directional indications (such as up, down, left, right, front, back...) involved in the embodiments of the present application, the directional indications are only used to explain the relative positional relationship and movement conditions between components in a specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indications will also change accordingly.

[0024] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present application, the descriptions of "first", "second", etc. are only for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include at least one such feature. In addition, the technical solutions between various embodiments can be combined with each other, but it must be based on the fact that those of ordinary skill in the art can implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the protection scope required by the present application.

[0025] Please refer to Figure 1 , Figure 1 which is a schematic structural diagram of the first embodiment of the optical measurement device provided by the present application. As Figure 1 shown, in this embodiment, the optical measurement device includes a sheath flow assembly 10, a laser assembly 20, a fluorescence receiving assembly (not shown in the figure), and a first shaping assembly 510.

[0026] The sheath flow assembly 10 is used to test the sample flow to be measured; the laser assembly 20 is disposed on one side of the sheath flow assembly 10 and includes at least two lasers with different wavelength ranges, which are used to emit laser light to the sheath flow assembly 10 so that the laser light irradiates the sample flow to be measured and excites the sample flow to be measured to generate a plurality of fluorescence beams with different wavelengths; the fluorescence receiving assembly is disposed on the side of the sheath flow assembly 10 away from the laser assembly 20, and the fluorescence detection assembly is used to receive the fluorescence beams; the first shaping assembly 510 is located between the fluorescence receiving assembly and the sheath flow assembly 10 and is used to homogenize and shape the fluorescence beams to reduce the color difference between the fluorescence beams with different wavelengths excited by at least two lasers with different wavelength ranges.

[0027] Specifically, the sheath flow assembly 10 may include a flow chamber, an inner sheath inlet, and an outer sheath inlet. The inner sheath inlet is used to inject the sample flow to be measured, and the outer sheath inlet is used to inject the sheath fluid. A detection channel for a single particle to be measured is provided inside the flow chamber. The sheath fluid wraps the sample flow to be measured and causes the sample flow to be measured to converge and then pass through the detection channel in sequence. The laser assembly 20 emits laser light to a certain position of the detection channel so that the particles to be measured in the sample flow to be measured emit fluorescence and scattered light under the irradiation of the laser. The optical measurement device collects the excited fluorescence through the fluorescence receiving assembly to obtain the detection result of the sample flow to be measured.

[0028] Among them, the laser assembly 20 includes at least two lasers with different wavelength ranges. The multiple lasers are arranged on the same side of the sheath flow assembly. The laser light emitted by different lasers is incident on the detection channel at the same angle, so that the laser light with different wavelength ranges can irradiate the particles to be measured at different positions of the detection channel. The laser irradiation direction of the laser assembly 20 in this embodiment is unified to reduce the restriction on the flow direction of the sample flow to be measured and reduce the internal space occupied by the laser optical path of the laser assembly 20. Moreover, the laser assembly 20 in this embodiment is provided with multiple lasers with different wavelength ranges to enable laser beams with different wavelengths to be used for quantitative analysis of different types of particles to be measured in the sample flow to be measured. The user can perform tests on multiple fluorescence channels based on a single test, improving the sample test efficiency.

[0029] The laser assembly 20 causes the particles to be measured to emit at least a first fluorescence beam and a second fluorescence beam with different wavelengths through at least two lasers with different wavelength ranges. When the sample flow to be measured is a sheath fluid sample prepared from a blood sample, the particles to be measured are substances such as antigens and antibodies in the blood. The fluorescence receiving assembly includes a first receiver 310 and a second receiver 320. The first receiver 310 is used to receive the first fluorescence beam, and the second receiver 320 is used to receive the second fluorescence beam. The optical measurement device is used to obtain the detection result of the sample flow to be measured through the intensity parameters of the first fluorescence beam and the second fluorescence beam. Further, the optical measurement device can perform classification analysis on the sample flow to be measured through at least one of the first fluorescence beam and the second fluorescence beam to divide the particles to be measured in the sample flow to be measured into classifications of multiple detection items; when the optical measurement device performs classification analysis through the first fluorescence beam, the optical measurement device can also perform quantitative analysis through the second fluorescence beam to obtain the concentration information or particle number information of the sample flow to be measured under the classification of this detection item.

[0030] In an alternative embodiment, the laser assembly 20 may include a first laser 210 and a second laser 220. The first laser 210 is configured to emit a first laser, and the second laser 220 is configured to emit a second laser. The wavelengths of the first laser and the second laser do not overlap. Exemplarily, the first laser 210 may provide a first laser with any wavelength within the range of 633 - 643 nm, such as 638 nm; the second laser 220 may provide a second laser with any wavelength within the range of 527 nm - 537 nm, such as 532 nm. Further, the laser assembly 20 may further include a second beam splitter 230 and a reflector. The first laser 210 is arranged at a 45° angle along the emission direction of the first laser, and the center of the reflector is aligned with the beam center of the first laser; the second laser 220 is arranged at a 45° angle along the emission direction of the second laser, and the center of the second beam splitter 230 is aligned with the beam center. The reflector is configured to reflect almost all of the first laser emitted by the first laser 210 to the second beam splitter 230, and the second beam splitter 230 is configured to allow a beam with a specific wavelength to pass through and reflect a beam with a wavelength other than the specific wavelength. Exemplarily, the second beam splitter 230 may allow a beam with a wavelength above 600 nm to pass through and reflect a beam with a wavelength below 600 nm, so that the first laser emitted by the first laser 210 and the second laser emitted by the second laser 220 are combined and projected into the sheath flow assembly 10 after passing through the second beam splitter 230.

[0031] Since the optical measurement device in this embodiment emits lasers through two lasers with different wavelength ranges, the degree of light beam offset after refraction of laser beams with different wavelengths through the same lens is relatively large. For example, since the wavelength of the first laser is 638 nm and the wavelength of the second laser is 532 nm, the difference in the laser wavelengths of the two is relatively large, resulting in a relatively large degree of offset between the light beams of the first laser and the second laser when they pass through the same lens for light transmission. The positions where the first laser and the second laser irradiate on the sheath flow assembly 10 do not coincide, resulting in a relatively large color difference between the first fluorescence beam and the second fluorescence beam. Therefore, in this embodiment, the fluorescence receiving assembly is arranged in the fluorescence emission direction of the sheath flow assembly 10, and the first shaping assembly 510 is arranged between the fluorescence receiving assembly and the sheath flow assembly 10. The first shaping assembly 510 is configured to perform micro - shaping on the fluorescence beams emitted by the sheath flow assembly 10 to adjust the angles of fluorescence beams with different wavelengths, etc., so that the shaped fluorescence beams are superimposed on each other and cancel the non - uniformity between the beams, thereby reducing the color difference between fluorescence beams with different wavelengths and further reducing the crosstalk between fluorescence beams with different wavelengths.

[0032] In an embodiment of the present application, the sheath flow assembly 10 of the optical measurement device is used to test a sample flow to be measured; the laser assembly 20 is disposed on one side of the sheath flow assembly 10 and includes at least two lasers with different wavelength ranges, which are used to emit laser light to the sheath flow assembly 10 so that the laser light irradiates the sample flow to be measured and excites the sample flow to be measured to generate a plurality of fluorescence beams with different wavelengths; the fluorescence receiving assembly is disposed on the side of the sheath flow assembly 10 away from the laser assembly 20, and the fluorescence detection assembly is used to receive the fluorescence beams; the first shaping assembly 510 is located between the fluorescence receiving assembly and the sheath flow assembly 10 and is used to homogenize and shape the fluorescence beams. In the above manner, the first shaping assembly 510 is disposed between the fluorescence receiving assembly and the sheath flow assembly 10, and the fluorescence beams excited by the sheath flow assembly 10 are homogenized and shaped by the first shaping assembly 510 to reduce the color difference between the fluorescence beams with different wavelengths excited by at least two lasers with different wavelength ranges, improve the signal-to-noise ratio of the fluorescence receiving assembly, and further improve the accuracy of optical measurement.

[0033] In one embodiment, the first shaping assembly 510 includes at least one microlens array 511 and a collection lens 512. The fluorescence receiving assembly includes a first beam splitter 360. The fluorescence beams excited by the sheath flow assembly 10 sequentially pass through the microlens array 511 and the collection lens 512 and enter the first beam splitter 360. The microlens array 511 homogenizes and shapes the fluorescence beams through a plurality of microlens sub-units. The collection lens 512 is used to collect the shaped fluorescence beams, and the first beam splitter 360 is used to split the fluorescence beams with different wavelengths.

[0034] Specifically, the first shaping assembly 510 may include at least one microlens array 511 and a collection lens 512. The laser beam emitted by the laser assembly 20 is incident on the sheath flow assembly 10 in a first direction, and the sample flow to be measured flows in a second direction in the detection area of the sheath flow assembly 10. The sheath flow assembly 10, the microlens array 511, the collection lens 512, the first beam splitter 360, and the fluorescence collection assembly 530 are sequentially arranged in the second direction, and the first direction is perpendicular to the second direction.

[0035] After the laser emitted by the laser assembly 20 irradiates the sample flow to be measured of the sheath flow assembly 10, the particles to be measured in the sample flow to be measured are excited to emit a fluorescence beam, and the fluorescence beam is emitted along the second direction and hits the microlens array 511. The microlens array 511 can be provided with a plurality of microlens sub-units, and the plurality of microlens sub-units are arranged in a periodic pattern or a random dot pattern. Each microlens sub-unit is used to focus and shape a part of the fluorescence beam so that the fluorescence beam is divided into a plurality of small beams. The small beams are refocused through each microlens sub-unit to form a focal point arranged in an array. The refocused plurality of small beams are superimposed on each other to cancel the non-uniformity between the beams, and the achromatic effect is good. The collection lens 512 is used to collect the shaped fluorescence beam and enhance it to improve the signal-to-noise ratio of the fluorescence receiving assembly.

[0036] In an alternative embodiment, please refer to Figure 2 , Figure 2 is Figure 1 a schematic structural diagram of an embodiment of the first shaping component in Figure 2 As shown, the first shaping component 510 may include two microlens arrays 511. The first microlens array 511 receives the incident fluorescence beam. The center of the second microlens array 511 is aligned with the center of the first microlens array 511. The microlens sub-units of the second microlens array 511 and the second microlens array 511 are symmetrically arranged or approximately symmetric, so that when the fluorescence beam is incident on the first microlens array 511, it can be regarded as a beam cluster array corresponding to the microlens array 511. The fluorescence beams emitted by the second microlens array 511 are superimposed on each other based on the symmetry of the array arrangement to cancel the non-uniformity of the fluorescence beams between different wavelengths, and the chromatic aberration elimination effect is good to avoid crosstalk between the fluorescence beams of different wavelengths.

[0037] Optionally, the distance between the microlens array 511 and the light-emitting surface of the sheath flow assembly 10 is 4 to 8 mm, and the diameter of the microlens sub-unit is 2.5 to 2.8 mm.

[0038] Specifically, the fluorescence beam directly enters the microlens array 511 through the light-emitting surface of the sheath flow assembly 10, and the distance between the light-emitting surface of the microlens array 511 and the microlens array 511 is 4 to 8 mm. For example, the distance between the light-emitting surface of the microlens array 511 and the microlens array 511 can be 4, 5, 6, 7 or 8 mm.

[0039] Understandably, when the distance between the microlens array 511 and the light-emitting surface of the sheath flow assembly 10 is less than 4 mm, the distance between the microlens array 511 and the light-emitting surface of the sheath flow assembly 10 is too close, or when the distance between the microlens array 511 and the light-emitting surface of the sheath flow assembly 10 is greater than 8 mm, the distance between the microlens array 511 and the light-emitting surface of the sheath flow assembly 10 is too far, both of which will result in a decrease in the stability of the fluorescence beam formed by the sheath flow assembly 10 at the light-emitting surface, a relatively large angular deviation of the fluorescence beam emitted by the microlens array 511, and poor achromatic aberration effect of the first shaping assembly 510. When the distance between the microlens array 511 and the light-emitting surface of the sheath flow assembly 10 is between 4 and 8 mm, the fluorescence beam formed by the light-emitting surface of the sheath flow assembly 10 can be projected onto the microlens array 511 at an appropriate incident angle to ensure the optical path adjustment accuracy and stability and ensure the achromatic aberration effect of the first shaping assembly 510.

[0040] Among them, the size of the microlens array 511 can be but is not limited to 10 mm * 9.8 mm * 1.6 mm, and the diameter or aperture of each microlens sub-unit in the microlens array 511 can be 2.5, 2.6, 2.7 or 2.8 mm. At this size of the microlens array 511, the microlens array 511 can allow as many fluorescence beams as possible to pass through, and at this diameter of the microlens sub-unit, the beam transmission effect between each lens is good, and the achromatic aberration effect of the fluorescence beams of different wavelengths is good.

[0041] Optionally, please refer to Figure 3 , Figure 3 which is a schematic structural diagram of the second embodiment of the optical measurement device provided by this application. As Figure 3 shown, the optical measurement device further includes a collection assembly 530. The collection assembly 530 is disposed between the first shaping assembly 510 and the sheath flow assembly 10. The collection assembly 530 is used to focus the fluorescence beam excited by the sheath flow assembly 10 and transmit the focused fluorescence beam to the first shaping assembly 510; among them, the distance between the microlens array 511 and the light-emitting surface of the collection assembly 530 is 4 - 8 mm, and the diameter of the microlens sub-unit is 2.5 - 2.8 mm.

[0042] In one embodiment, the collection component 530 consists of a focusing lens, which is used to collect the fluorescence beam excited by the sheath flow component 10 in a focused manner and transmit the optical information of the fluorescence beam to the first shaping component 510, so that as many fluorescence beams as possible are restored and transmitted to the rear optical path. In another embodiment, the collection component 530 includes a lens group composed of multiple focusing lenses, which is used to focus the fluorescence beam excited by the sheath flow component 10 and collimate the focused fluorescence beam, so as to transmit the collimated fluorescence beam to the first shaping component 510. Therefore, by setting the collection component 530, the optical measurement device of this embodiment can collect and restore as many fluorescence beams excited by the sheath flow component 10 as possible, and transmit the fluorescence beam to the first shaping component 510 to enhance the fluorescence beam of the sheath flow component 10, improve the signal-to-noise ratio of the optical measurement device, and further improve the anti-interference performance of the optical measurement device.

[0043] Specifically, in this embodiment, the optical measurement device sets the collection component 530 between the first shaping component 510 and the sheath flow component 10 to enhance the fluorescence beam. Similarly, the distance between the microlens array 511 and the light-emitting surface of the collection component 530 is 4-8 mm. For example, the distance between the microlens array 511 and the light-emitting surface of the collection component 530 can be 4, 5, 6, 7, or 8 mm. The fluorescence beam formed by the light-emitting surface of the sheath flow component 10 can be projected onto the microlens array 511 at an appropriate incident angle to ensure the achromatic aberration effect between the fluorescence beams.

[0044] In one embodiment, the laser beam emitted by the laser component 20 is incident on the sheath flow component 10 along the first direction and excites the sample flow to be measured to generate a fluorescence beam and a scattered beam. The fluorescence receiving component includes a first filter 340, a first receiver 310, a second filter 350, and a second receiver 320. The first receiver 310 is used to receive the fluorescence beam filtered by the first filter 340, and the second receiver 320 is used to receive the fluorescence beam filtered by the second filter 350. The optical measurement device further includes a third filter 420 and a scattered light receiver 410, and the scattered light receiver 410 is used to receive the scattered beam filtered by the third filter 420.

[0045] Specifically, the laser beam emitted by the laser assembly 20 is incident on the sheath flow assembly 10 in the first direction, and the sample flow to be measured flows in the second direction in the detection area of the sheath flow assembly 10, so that the sample flow to be measured is excited to generate a fluorescence beam and a scattered beam. The scattered beam generated by the sheath flow assembly 10 continues to be incident on the third filter 420 in the first direction. The third filter 420 is used to filter out the light beam with a specific wavelength and cut off the light beam outside the specific wavelength, so that the scattered light receiver 410 can receive the scattered beam with a specific wavelength and perform a counting test on the particles to be measured based on the intensity parameter of the scattered beam.

[0046] The fluorescence receiving assembly further includes at least one first beam splitter 360. The fluorescence beam generated by the sheath flow assembly 10 is incident on the first beam splitter 360 in the second direction. The first beam splitter 360 is used to transmit the fluorescence beam in a certain wavelength range and reflect the fluorescence beam in other wavelength ranges. In an optional implementation manner, when the fluorescence receiving assembly is only provided with one first beam splitter 360, a first receiver 310, and a second receiver 320, the first receiver 310 is arranged in the reflection direction of the first beam splitter 360, and the second receiver 320 is arranged in the transmission direction of the first beam splitter 360. The first receiver 310 is used to receive the fluorescence beam reflected by the first beam splitter 360, and the second receiver 320 is used to receive the fluorescence beam transmitted by the first beam splitter 360.

[0047] In other embodiments, please refer to Figure 4 , Figure 4 which is a schematic structural diagram of the third embodiment of the optical measurement device provided by the present application. As Figure 4 shown, the fluorescence receiving assembly may be provided with n receivers and n - 1 first beam splitters 360. The first first beam splitter 360 is used to receive the fluorescence beam incident in the second direction. The first receiver 310 is arranged in the reflection direction of the first beam splitter 360. The second first beam splitter 360 is arranged in the transmission direction of the first first beam splitter 360. The second receiver 320 is arranged in the reflection direction of the second first beam splitter 360, and so on, until the nth receiver is arranged in the transmission direction of the (n - 1)th first beam splitter 360, so that the optical measurement device can realize the measurement of multiple fluorescence detection channels, which will not be elaborated here.

[0048] Further, please refer to Figure 5 , Figure 5 which is a schematic structural diagram of the fourth embodiment of the optical measurement device provided by the present application. As Figure 5As shown, the beam splitter is used to change the outgoing direction of the fluorescence beam, so that the first filter 340, the second filter 350, and the third filter 420 are all used to receive the fluorescence beam outgoing in the first direction. At this time, in order to reduce the mutual crosstalk between the fluorescence beams of different wavelengths in the same direction, at least one of the first filter 340, the second filter 350, and the third filter 420 has a first distance from the adjacent receiver of the filter close to the sheath flow assembly 10 in the first direction. It can be understood that the adjacent receiver of the filter is the receiver with the closest distance in the opposite direction of the filter in the first direction, and the first distance is not 0, that is, at least one of the first filter 340, the second filter 350, and the third filter 420 is arranged at intervals from its adjacent receiver. Among them, a projection plane can be set in the first direction, and the first distance can be the distance between the filter and the receiver when they are projected onto this projection plane.

[0049] When the sample flow to be measured in the sheath flow assembly 10 is excited to generate a fluorescence beam, scattered light will also be generated in other directions except the second direction; moreover, since there are multiple optical devices inside the optical measurement device, the fluorescence (stray light) that is not required by the receiver inside may also be projected onto the receiver, resulting in excessive noise in the fluorescence measurement. In this embodiment, by arranging the filter at intervals from its adjacent receiver, the filter can be used to filter the stray light that may appear in the first direction, reduce the crosstalk between the fluorescence beams of different wavelengths, and improve the signal-to-noise ratio of the optical measurement device. And / or, there is a second distance between the scattered light receiver and the first receiver, and between the scattered light receiver and the second receiver in the second direction. The second direction is the flow direction of the sample flow to be measured in the sheath flow assembly 10, and the second direction is perpendicular to the first direction. The second distance is not 0, so that the stray light of the scattered light receiver in the second direction will not crosstalk with the fluorescence of the first receiver and the second receiver, and improve the signal-to-noise ratio of the fluorescence receiving assembly.

[0050] Optionally, the distance between the scattered light receiver 410 and the first filter 340 in the first direction is the first distance; and / or, the distance between the first receiver 310 and the second filter 350 in the first direction is the first distance, and the first distance is 30 - 80 mm.

[0051] Specifically, when the fluorescence receiving component includes three receivers (a first receiver 310, a second receiver 320, and a third receiver 330) and two filters, both the first receiver 310 and the second receiver 320 are used to receive the fluorescence beam emitted in the first direction, and the third receiver 330 is used to receive the fluorescence beam emitted in the second direction. At this time, when the sheath flow component 10, the first filter 340, the second filter 350, the third filter 420, the first receiver 310, the second receiver 320, and the scattered light receiver 410 are projected onto the projection plane, the positional relationship among them is: the sheath flow component 10, the third filter 420, the scattered light receiver 410, the first filter 340, the first receiver 310, the second filter 350, and the second receiver 320.

[0052] It can be understood that since the scattered light receiver 410, the first receiver 310, and the second receiver 320 are all used to receive the fluorescence beam transmitted in the first direction, setting the scattered light receiver 410 before the first filter 340 can ensure that the scattered light receiver 410 completely receives the scattered light beam, and the first filter 340 can filter the forward-scattered stray light to reduce the crosstalk degree of the fluorescence beam of the first receiver 310. Similarly, setting the first receiver 310 on the front side of the second filter 350 can filter the forward-scattered stray light through the first filter 340 and filter the stray fluorescence before entering the first receiver 310 through the second filter 350, reducing the crosstalk degree of the fluorescence beam of the second receiver 320 and improving the signal-to-noise ratio of the fluorescence receiving component.

[0053] Specifically, the first distance can be related to the fluorescence transmission optical path and the receiving target surface of the receiver. The first distance can be but is not limited to 30 - 80 mm. For example, the first distance can be 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 mm. In the optical path of this embodiment, by limiting the first distance between the filter and its adjacent receiver to 30 - 80 mm, the filter can effectively filter the stray light before entering the adjacent receiver, with good filtering effect and low crosstalk degree, further improving the signal-to-noise ratio of the fluorescence receiving component.

[0054] Optionally, the wavelength range of the first fluorescence beam received by the first receiver 310 is less than the wavelength range of the second fluorescence beam received by the second receiver 320, and / or the scattered light receiver 410 is used to receive the scattered light beam and obtain the forward-scattered light intensity of the scattered light beam.

[0055] Specifically, the wavelength ranges of the first fluorescence beam received by the first receiver 310 and the second fluorescence beam received by the second receiver 320 do not cross, and the wavelength ranges that can pass through the first filter 340 and the second filter 350 also do not cross. The wavelength range of the first fluorescence beam received by the first receiver 310 is less than the wavelength range of the second fluorescence beam received by the second receiver 320, such that the distance between the first receiver 310 and the sheath flow assembly 10 is less than the distance between the second receiver 320 and the sheath flow assembly 10. Since the signal of the fluorescence beam is weak and is prone to loss during the optical path transmission, therefore, the first receiver 310 with a shorter transmission process and a closer distance is set to receive the fluorescence beam with a shorter wavelength, which can reduce the loss degree of the fluorescence beam and enable the fluorescence beam to accurately reflect the characteristics of the particles to be measured. The scattered light receiver 410 is used to receive the scattered beam and obtain the forward scattered light intensity of the scattered beam, such that the optical measurement device can perform counting analysis on the particles to be measured in the sample flow to be measured through the forward scattered light intensity of the sample flow to be measured.

[0056] Optionally, the fluorescence receiving assembly is disposed on one side of the sheath flow assembly 10 along the second direction. The fluorescence beam includes a first fluorescence beam and a second fluorescence beam with different wavelengths. The first filter 340 is used to filter the first fluorescence beam so that the filtered first fluorescence beam is incident on the first receiver 310 along the first direction, and the second filter 350 is used to filter the second fluorescence beam so that the filtered second fluorescence beam is incident on the second receiver 320 along the first direction. That is, the first receiver 310, the second receiver 320, and the scattered light receiver 410 are all used to receive the beam incident along the first direction.

[0057] The laser assembly 20, the sheath flow assembly 10, the third filter 420, and the scattered light receiver 410 are sequentially disposed along the first direction; when the third filter 420, the first filter 340, and the second filter 350 are projected in the first direction, the projection positions of the third filter 420, the first filter 340, and the second filter 350 are arranged in sequence; the scattered light receiver 410, the first receiver 310, and the second receiver 320 are spaced apart along the second direction. Specifically, along the second direction, the third filter 420 and the scattered light receiver 410 are disposed on the first plane, the second filter 350 and the second receiver 320 are disposed on the second plane, the first filter 340 and the first receiver 310 are disposed on the third plane, the height of the first plane is less than the height of the third plane, and the height of the third plane is less than the height of the second plane. In this embodiment, multiple receivers and filters of the optical measurement device are arranged in an orderly manner inside the device, such that the multiple optical paths of the optical measurement device are reasonably arranged, improving the stability of the optical measurement device.

[0058] Optionally, the second distance is 10 to 30 mm; for example, the second distance can be 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, or 30 mm.

[0059] Wherein, in the second direction, there is a first height difference between the scattered light receiver 410 and the first receiver 310, and a second height difference between the scattered light receiver 410 and the second receiver 320. The first height difference and the second height difference may refer to the height difference between the center points of the light-receiving surfaces of the corresponding receivers, and the second height difference is greater than the first height difference.

[0060] In this embodiment, by defining the second distance between the scattered light receiver 410, the first receiver 310, and the second receiver 320, the crosstalk of the scattered light in the second direction before being received by the scattered light receiver 410 on the received fluorescence beam is reduced, the signal-to-noise ratio of the fluorescence receiving component is improved, and thus the accuracy of optical measurement is improved.

[0061] In one embodiment, the fluorescence receiving component includes a plurality of first beam splitters 360. The plurality of first beam splitters 360 are used to separate the fluorescence beam at different wavelengths. The plurality of first beam splitters 360 are arranged in sequence along the optical path of the fluorescence beam, so that the fluorescence beams of different wavelengths are used for classification testing and / or quantitative testing of the sample flow to be measured.

[0062] Specifically, the first beam splitter 360 is used to change the transmission direction and position of the fluorescence beams of different wavelengths to separate the fluorescence beams of different wavelengths. In an optional embodiment, the fluorescence receiving component includes 2 first beam splitters 360, a first receiver 310, a second receiver 320, and a third receiver 330. The sheath flow component 10 emits a fluorescence beam in the second direction to the first first beam splitter 360. The first first beam splitter 360 is used to reflect the fluorescence beam of the first wavelength, so that the fluorescence beam of the first wavelength is transmitted in the first direction into the first receiver 310; the first first beam splitter 360 is also used to transmit the fluorescence beam other than the first wavelength to the second first beam splitter 360. The second first beam splitter 360 is used to reflect the fluorescence beam of the second wavelength, so that the fluorescence beam of the second wavelength is transmitted in the first direction into the second receiver 320, and the fluorescence beam other than the second wavelength continues to be transmitted in the transmission direction of the second first beam splitter 360 to the third receiver 330.

[0063] Each particle to be measured is conjugated with at least one substance to be measured. Quantitative analysis can be performed by analyzing the concentration of the substance to be measured in the sample flow to be measured through one of the receivers. The substance to be measured can be various antigens or antibodies in a blood sample, such as antigen A / B / C / D / etc., or other substances that need to be classified and counted. Each substance to be measured corresponds to a classification detection item. Classification analysis can be performed by analyzing the categories of different substances to be measured in the sample flow to be measured through receivers other than the receiver used for quantitative analysis. The detection results output after the optical measurement device measures the sample to be measured include the quantitative (concentration) information corresponding to each classification detection item respectively.

[0064] Exemplarily, when the fluorescence receiving component includes a first receiver, a second receiver, and a third receiver, each particle to be measured is conjugated with at least one substance to be measured and magnetic beads. The magnetic beads are coated with a second fluorescent substance and a third fluorescent substance with intensity levels corresponding to the substance. The substance to be measured can be a substance labeled with a first fluorescent substance. The first receiver is used to quantitatively analyze the concentration of the substance to be measured based on the fluorescence intensity of the first fluorescent substance. The second receiver is used to analyze the categories of different substances to be measured based on the second fluorescent substances with different intensity levels. The third receiver is used to analyze the categories of different substances to be measured based on the third fluorescent substances with different intensity levels.

[0065] In other embodiments, the optical measurement device can also be used for joint detection of different detection items through fluorescence beams of multiple wavelengths by continuously adding a first beam splitter 360 and receivers. Among them, the number of joint detections corresponds to the number of intensity levels of the second fluorescent substance and the third fluorescent substance coated on the magnetic beads; the second fluorescent substance with N intensity levels and the third fluorescent substance with M intensity levels can achieve joint detection of N*M substances to be measured. The optical measurement device adjusts the intensity parameters of different substances to output the quantitative information corresponding to at most N*M classification detection items of the sample to be measured.

[0066] In other embodiments, the fluorescence receiving component can also replace the first beam splitter 360 with a beam splitting optical fiber to achieve beam splitting of the fluorescence beam, reduce the complexity of the debugging process, and reduce problems such as possible errors that may occur during the assembly process for the user.

[0067] In one embodiment, the optical measurement device further includes a second shaping component 520. The second shaping component 520 is disposed between the laser component 20 and the sheath flow component 10. The second shaping component 520 is used to receive the laser beam emitted by the laser component 20 and collect and collimate the laser beam to perform flat-topping processing on the laser beam.

[0068] Specifically, by disposing a second shaping component 520 at the front end of the sheath flow component 10, the laser beam emitted by the laser component 20 is collected and collimated, and the laser beam is subjected to spot diffusion and color mixing to achieve the flat-topping processing of the laser beam, so that the laser beam emitted by the second shaping component 520 has uniform light output, the first laser and the second laser of the laser component 20 coincide at the spot position of the sheath flow component 10, reduce the crosstalk between the fluorescence beams of different wavelengths acquired by the fluorescence receiving component, improve the consistency between different fluorescence beams, and further improve the signal-to-noise ratio of the optical measurement system.

[0069] The above are only the embodiments of the present application, and do not limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made by using the content of the specification and drawings of the present application, or directly or indirectly applied in other related technical fields, shall be equally included in the patent protection scope of the present application.

Claims

1. An optical measurement device, characterized in that, Comprising: A sheath flow component for testing a sample flow to be tested; A laser component disposed on one side of the sheath flow component, including at least two lasers with different wavelength ranges, for emitting laser light to the sheath flow component, so that the laser light irradiates the sample flow to be tested and excites the sample flow to be tested to generate a plurality of fluorescent beams with different wavelengths; A fluorescence receiving component disposed on the side of the sheath flow component away from the laser component, and the fluorescence detection component is used to receive the fluorescent beams; A first shaping component located between the fluorescence receiving component and the sheath flow component, for homogenizing and shaping the fluorescent beams to reduce the color difference between the fluorescent beams excited by the at least two lasers with different wavelength ranges.

2. The optical measurement device according to claim 1, characterized in that, The first shaping component includes at least one microlens array and a collecting lens. The fluorescence receiving component includes a first beam splitter. The fluorescent beams excited by the sheath flow component sequentially pass through the microlens array and the collecting lens and enter the first beam splitter. The microlens array homogenizes and shapes the fluorescent beams through a plurality of microlens sub-units. The collecting lens is used to collect the shaped fluorescent beams, and the first beam splitter is used to split the fluorescent beams with different wavelengths.

3. The optical measurement device according to claim 2, wherein The distance between the microlens array and the light-emitting surface of the sheath flow component is 4 to 8 mm, and the diameter of the microlens sub-unit is 2.5 to 2.8 mm.

4. The optical measurement device according to claim 2, characterized in that The optical measurement device further includes a collecting component disposed between the first shaping component and the sheath flow component. The collecting component is used to focus the fluorescent beams excited by the sheath flow component and transmit the focused fluorescent beams to the first shaping component; wherein, the distance between the microlens array and the light-emitting surface of the collecting component is 4 to 8 mm, and the diameter of the microlens sub-unit is 2.5 to 2.8 mm.

5. The optical measurement device according to claim 1, characterized in that, The laser beam emitted by the laser component is incident on the sheath flow component along a first direction and excites the sample flow to be tested to generate the fluorescent beams and scattered beams. The fluorescence receiving component includes a first filter, a first receiver, a second filter, and a second receiver. The first receiver is used to receive the fluorescent beams filtered by the first filter, and the second receiver is used to receive the fluorescent beams filtered by the second filter. The optical measurement device further includes a third filter and a scattered light receiver, and the scattered light receiver is used to receive the scattered beams filtered by the third filter; Wherein, at least one of the first filter, the second filter, and the third filter has a first distance from the adjacent receiver of the filter close to the sheath flow component in the first direction, and / or, the scattered light receiver has a second distance from the first receiver and the second receiver respectively in a second direction, and the second direction is the flowing direction of the sample flow to be tested of the sheath flow component.

6. The optical measurement device according to claim 5, characterized in that The distance between the scattered light receiver and the first filter in the first direction is the first distance; and / or, the distance between the first receiver and the second filter in the first direction is the first distance, and the first distance is 30 to 80 mm.

7. The optical measurement device according to claim 5, characterized in that, The wavelength range of the first fluorescence beam received by the first receiver is less than the wavelength range of the second fluorescence beam received by the second receiver, and / or, the scattered light receiver is configured to receive the scattered beam and obtain the forward scattered light intensity of the scattered beam.

8. The optical measurement device according to claim 5, characterized in that, The fluorescence receiving assembly is disposed on one side of the sheath flow assembly along the second direction. The fluorescence beam includes a first fluorescence beam and a second fluorescence beam with different wavelengths. The first filter is configured to filter the first fluorescence beam so that the filtered first fluorescence beam is incident on the first receiver along the first direction. The second filter is configured to filter the second fluorescence beam so that the filtered second fluorescence beam is incident on the second receiver along the first direction. Wherein, the laser assembly, the sheath flow assembly, the third filter, and the scattered light receiver are sequentially arranged along the first direction. The projection positions of the third filter, the first filter, and the second filter are sequentially arranged in the first direction. The scattered light receiver, the first receiver, and the second receiver are spaced apart in the second direction.

9. The optical measurement device according to claim 5, characterized in that The second distance is 10 to 30 mm.

10. The optical measurement device according to claim 1, characterized in that, The fluorescence receiving assembly includes a plurality of first beam splitters. The plurality of first beam splitters are configured to separate the fluorescence beam at different wavelengths. The plurality of first beam splitters are sequentially arranged along the optical path of the fluorescence beam so that the fluorescence beams with different wavelengths are used for classification testing and / or quantitative testing of the sample flow to be measured.

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