Cuvette assemblies, flow cells equipped with cuvette assemblies, and sample processors containing cuvette assemblies or flow cells.
The cuvette assembly with a truncated reflector and focusing lens, combined with a flow cell having a smooth channel and degassing mechanism, addresses the inefficiency in collecting light signals, improving optical detection and sorting performance.
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Patents
- Current Assignee / Owner
- BECKMAN COULTER BIOTECHNOLOGY (SUZHOU) CO LTD
- Filing Date
- 2021-11-01
- Publication Date
- 2026-04-23
AI Technical Summary
Existing flow cells and cuvette assemblies in sample processors face challenges in efficiently collecting weak side-scattered light and fluorescence signals from biological particles, which hampers optical detection performance.
A cuvette assembly with a reflector having a truncated spherical surface and a focusing lens, along with an aspherical lens, is designed to enhance the collection of scattered and fluorescent light, while a flow cell with a base featuring a smooth channel and degassing mechanism ensures stable fluid flow and bubble removal.
The design significantly improves the efficiency of collecting side-scattered and fluorescent light, enhancing optical detection performance and sorting capabilities of the sample processor.
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Abstract
Description
Technical Field
[0001] The present invention relates to a flow cell and cuvette assembly for a sample processor such as a flow cell sorter / analyzer, and the present invention also relates to a corresponding sample processor.
Background Art
[0002] The content of this section only provides background information related to the present disclosure, which is not necessarily prior art.
[0003] Sample processors are often used to analyze and / or sort samples such as microsomes or cells. A flow cell sorter / analyzer is an instrument for performing multi-parameter and rapid qualitative / quantitative analysis and / or sorting of single cells or other biological particles in a single row in a high-speed linear flow state based on flow cytometry. It usually includes a flow cell and fluid system, a light source and optical system, a sample analysis and / or sorting system, etc. The flow cell includes a sample detection area such as a cuvette with a sample detection channel. Fluorescent staining is performed on the cells or other particles to be tested, and a sample suspension is prepared. The droplets of the sample suspension are encapsulated by the sheath liquid, then pass through the sample detection area of the flow cell, are irradiated by a light source (generally a laser light source) in the sample detection area, and generate side scatter light and fluorescence signals that reflect the information of the sample. The optical signals are collected by the optical system, then converted and amplified, and processed and analyzed by a signal processing device.
[0004] The sample sorting system can, for example, charge the sample droplets flowing out of the flow cell with different positive and negative charges according to the results of signal processing and analysis, so that the sample droplets are deflected under the action of a high-voltage electric field and collected in different collection containers, thus realizing the sorting of the sample.
[0005] The side-scattered light and fluorescence signals emitted by cells or other biological particles within the sample detection area of a flow cell after laser irradiation are relatively weak and difficult to collect. Therefore, it is desirable to design cuvette assemblies and corresponding flow cells that can improve the efficiency of collecting scattered and fluorescent light. [Overview of the project] [Means for solving the problem]
[0006] This section provides a general overview of the disclosure, rather than being a comprehensive disclosure of the entire scope or all features of the disclosure.
[0007] The object of the present invention is to provide a cuvette assembly for a sample processor having high efficiency in collecting scattered and fluorescent light.
[0008] Another object of the present invention is to provide a flow cell for a sample processor that can improve the optical detection performance of the sample processor, in particular, the efficiency of collecting scattered and fluorescent light.
[0009] Another object of the present invention is to provide a sample processor having improved optical detection performance.
[0010] According to one aspect of the present disclosure, a cuvette assembly for a sample processor is provided. The cuvette assembly includes a cuvette body and a reflector. The cuvette body is rectangular in shape and includes a sample detection channel that penetrates the cuvette body perpendicularly. The cuvette body has a long side and a short side in a horizontal cross-section. The reflector has a flat surface attached to a first side extending along one of the long sides of the cuvette body and a spherical surface opposite the flat surface, with the lower half cut off. The reflector is coplanar with the lower surface of the cuvette body, and the center of the sphere of the spherical surface is placed within the sample detection channel, and the reflector is positioned to extend along the long side and beyond the short side.
[0011] Partially cutting off the lower half of the spherical surface of the reflector facilitates subsequent sample sorting and other processing. The length of the reflector exceeds the length of the cuvette body, which expands the area of the reflector that can receive lateral scattered and fluorescent light, thereby significantly improving the efficiency of collecting lateral scattered and fluorescent light.
[0012] In some examples of the present disclosure, the cuvette assembly may further include a focusing lens attached to a second side extending along one of the shorter sides of the cuvette body, the focusing lens focusing incident light to the center of the sphere on the spherical surface of the reflector.
[0013] In some examples of the present disclosure, the cuvette assembly may further include an aspherical lens mounted on a third side opposite a first side of the cuvette body, the aspherical lens shaping a focused spot formed by reflection from a reflector and focusing the focused spot into a signal detection device for detecting an optical signal.
[0014] In some examples of the present disclosure, an aspherical lens includes a molded portion for shaping and an outer frame surrounding the molded portion, wherein the central portion of the molded portion is thicker than the peripheral portion.
[0015] In some examples provided herein, the sample detection channel has a square cross-section.
[0016] According to another aspect of this disclosure, a flow cell for a sample processor is provided. The flow cell comprises a frame, a flow cell body, and a nozzle assembly. The flow cell body is fixed to the frame, and the flow cell body comprises a base and a cuvette assembly according to the above aspect. The cuvette assembly is located below the base. Samples from a sample line and fluids from a fluid line are concentrated in the base and flow into a sample detection channel in the cuvette assembly. The nozzle assembly is located at the outlet of the sample detection channel and has a nozzle for discharging the sample in the sample detection channel in a given mode.
[0017] In some examples of the present disclosure, the base may include a vertical channel, which includes a smoothly transitioning cylindrical section and a tapered section having a smooth inner surface, such that a sample and fluid are concentrated within the channel, and the tapered channel is concentrically aligned with the sample detection channel.
[0018] In some examples of the present disclosure, the base may further include symmetrically arranged fluid ports communicating with fluid lines and channels, the fluid ports being higher than the outlet of the sample line.
[0019] In some examples of the present disclosure, the base may further include a degassing channel, one end of which communicates with the channel and the other end which is attached to a degassing device.
[0020] In some examples provided herein, the degassing channel is higher than the fluid port.
[0021] In some examples provided herein, the degassing channel is located at the top of the channel.
[0022] In some examples of this disclosure, the upper surface for defining the channel is tilted to induce the release of bubbles.
[0023] In some examples of the present disclosure, the flow cell body may further include a cover member located above the base, which causes the sample line to be concentrically aligned with the channel.
[0024] In some examples provided herein, the nozzle assembly is installed within the flow cell in a manner that allows it to be independently removed.
[0025] In some examples of the present disclosure, a cavity for housing a piezoelectric element is located on top of the base.
[0026] In some examples according to the present disclosure, the cavity for arranging the piezoelectric element is an annular cavity surrounding the sample line.
[0027] In some examples according to the present disclosure, the tail end of the sample line includes a rigid elongated member.
[0028] In some examples according to the present disclosure, the sample line extends into the tapered section of the channel.
[0029] In some examples according to the present disclosure, the outlet of the sample line is tapered.
[0030] According to yet another aspect of the present disclosure, there is provided a sample processor including the cuvette assembly and / or flow cell according to the above aspect.
[0031] In some examples according to the present disclosure, such a sample processor is a sample sorter. The present invention provides, for example, the following: (Item 1) A cuvette assembly for a sample processor, wherein the cuvette assembly is A cuvette body having the shape of a rectangular parallelepiped, wherein the cuvette body is provided with a sample detection channel that penetrates the cuvette body perpendicularly, and the cuvette body has a long side and a short side in a horizontal cross-section, reflector and Equipped with, The reflector has a flat surface attached to a first side surface extending along one of the long sides of the cuvette body, and a spherical surface opposite the flat surface, with the lower half cut off. A cuvette assembly in which the reflector is positioned such that it is coplanar with the lower surface of the cuvette body, the center of the sphere on the spherical surface is located within the sample detection channel, and the reflector extends along the long side and beyond the short side. (Item 2) [[ID= Nozzle assembly having a nozzle and Equipped with, The nozzle is located at the outlet of the sample detection channel and discharges the sample from the sample detection channel in a predetermined mode, forming a flow cell. (Item 7) The flow cell according to item 6, wherein the base comprises a vertical channel, the channel having a smooth inner surface and comprising a smoothly transitioning cylindrical section and a tapered section, the tapered section being concentrically aligned with the sample detection channel, so that the sample and the fluid can be concentrated within the channel. (Item 8) The base further comprises symmetrically arranged fluid ports communicating with the fluid line and the channel, wherein the fluid ports are higher than the outlet of the sample line, as described in item 7. (Item 9) The flow cell according to item 8, wherein the base further comprises a degassing channel, one end of the degassing channel communicating with the channel and the other end attached to a degassing device. (Item 10) The degassing channel is higher than the fluid port in the flow cell described in item 9. (Item 11) The degassing channel is a flow cell as described in item 10, located at the top of the channel. (Item 12) The flow cell according to item 11, wherein the upper surface for defining the channel is tilted to induce the release of bubbles. (Item 13) The flow cell according to item 7, wherein the flow cell body further comprises a cover member, the cover member located above the base, and the cover member causes the sample line to be concentrically aligned in the channel. (Item 14) The flow cell according to item 6, wherein the nozzle assembly is installed within the flow cell in a manner that allows it to be independently removed. (Item 15) The flow cell according to item 6, wherein a cavity for arranging a piezoelectric element is located at the top of the base. (Item 16) The flow cell according to item 15, wherein the cavity is an annular cavity surrounding the sample line. (Item 17) The flow cell described in item 6, wherein the tail end of the sample line is provided with a rigid, elongated member. (Item 18) The sample line extends into the tapered section of the channel, and the flow cell is as described in any one of items 7-17. (Item 19) The flow cell described in item 18, wherein the outlet of the sample line is tapered. (Item 20) A sample processor comprising a cuvette assembly as described in any one of items 1-5, and / or a flow cell as described in any one of items 6-19. (Item 21) The aforementioned sample processor is a sample sorter, as described in item 20. [Brief explanation of the drawing]
[0032] Throughout the following description, and by referring to the accompanying drawings, the features and advantages of one or more embodiments of this disclosure will be more easily understood in the accompanying drawings.
[0033] [Figure 1] Figure 1 is a three-dimensional schematic diagram of a sample processor according to one embodiment of the present disclosure. [Figure 2A] Figure 2A is a three-dimensional cross-sectional view of the sample processor obtained along the cross-sectional line AA in Figure 1. [Figure 2B] Figure 2B is a three-dimensional cross-sectional view of the sample processor obtained along the cross-sectional line BB in Figure 1. [Figure 3] Figure 3 is a partially enlarged view corresponding to the three-dimensional cross-sectional view shown in Figure 2A. [Figure 4-1] Figures 4A and 4B are three-dimensional views of a cuvette assembly according to one embodiment of the present disclosure, viewed from different angles. [Figure 4-2] Figure 4C is a top view of the cuvette assembly. [Figure 4-3] Figure 4D is a cross-sectional view of the cuvette assembly obtained along the cross-sectional line CC in Figure 4C. [Figure 5] Figures 5A and 5B are schematic optical path diagrams for collecting side-scattered and fluorescent light observed from above and to the side of the cuvette assembly, respectively. [Modes for carrying out the invention]
[0034] This disclosure will be described in detail below with reference to the accompanying drawings, through exemplary embodiments. In some of the accompanying drawings, similar reference numerals indicate similar parts and components. The following detailed description of this disclosure is for illustrative purposes only and is not in any way intended to limit this disclosure or its use or application. The embodiments described herein are not exhaustive and represent only a selection of several possible embodiments. Exemplary embodiments may be implemented in many different forms and should not be construed as limiting the scope of this disclosure. In some exemplary embodiments, well-known processes, well-known device structures, and well-known technologies may not be described in detail.
[0035] Figure 1 is a three-dimensional schematic diagram of a flow cell 1 for a sample processor according to one embodiment of the present disclosure. Figure 2A is a three-dimensional cross-sectional view of the flow cell 1 obtained along the cross-sectional line AA of Figure 1, and Figure 2B is a three-dimensional cross-sectional view of the flow cell 1 obtained along the cross-sectional line BB of Figure 1. The overall structure of the flow cell 1 will be described below with reference to Figures 1-2B.
[0036] As shown in Figures 1-2B, the flow cell 1 includes a flow cell body 10, a positioning member 20, a support member 30, a frame 40, and a nozzle assembly 50. The frame 40 serves as a fixed component of the flow cell 1 and is used to support and mount the other components. The flow cell body 10, the positioning member 20, and the support member 30 are all fixed to the frame 40, either directly or indirectly.
[0037] The various components of the flow cell 1 shown in Figures 1-2B are assembled in their fixed positions and working state. When the flow cell 1 is operating, the flow cell body 10 receives a sample from the sample line SL and a fluid such as sheath fluid from the fluid line FL. The sample and sheath fluid are concentrated within the flow cell body 10 and then discharged through the nozzle assembly 50. The sample is detected and analyzed as it flows through the flow cell body 10, and then the sample discharged from the nozzle assembly 50 is sorted based on the results of the detection and analysis.
[0038] The flow cell body 10 may include a base 100, a cover member 200, and a cuvette assembly 300, the cover member 200 and the cuvette assembly 300 being located above and below the base 100, respectively. The base 100 cooperates with the cover member 200 so that the sample and sheath fluid are concentrated and flow into the cuvette assembly 300, and the sample is detected within the cuvette assembly 300.
[0039] The base 100 comprises a vertical channel 110 and symmetrically arranged fluid ports 120a and 120b communicating with the fluid line FL, through which the sheath fluid flows into the channel 110 via the fluid ports 120a and 120b. The channel 110 has a smooth inner surface and includes a smoothly transitioning substantially cylindrical section and a tapered section, with the outlet 130 defined in the tapered section. The center of the cover member 200 comprises a through hole and an annular projection 210 extending perpendicularly downward around the through hole, the annular projection 210 being inserted into the substantially cylindrical section of the channel 110. The inner surface of the annular projection 210 defines a sample port, which allows the sample line SL to be inserted into the channel 110 and positioned concentrically with respect to the channel 110. The tail end of the sample line SL typically includes a rigid, elongated member, such as a sample probe, which is inserted into the channel 110 and positioned concentrically with respect to the channel 110. The outlet of the sample line SL generally extends into a tapered section of the channel 110, promoting the concentration of the sample and sheath fluid to form a uniform and stable sample flow. Preferably, the outlet of the sample line SL is designed as a tapered tip to reduce the influence of the sample on the stability of the laminar flow of the sheath fluid. The sample line SL is secured to the cover member 200 by screws. The base 100 and the cover member 200 are fixed by screws, and an O-ring 910 for sealing is positioned around the annular projection 210 between the base 100 and the cover member 200.
[0040] The sample from the sample line SL and the sheath fluid from the fluid line FL are concentrated within the channel 110. The smooth inner surface and simple structure of the channel 110 facilitate the concentration of the sample flow and sheath fluid flow, forming a uniform and stable laminar flow, thus avoiding fluid disturbances or dead flow zones and reducing the possibility of bubble accumulation and adhesion. Compared to existing flow cytometers available in [location], the channel 110 reduces the volume and surface area of the channel, thereby minimizing the possibility of bubble adhesion to the inner surface of the channel 110. In addition, the smaller volume of the channel 110 increases the fluid velocity, thereby facilitating bubble removal.
[0041] The fluid ports 120a and 120b are positioned sufficiently high relative to the outlet of the sample line SL to ensure that the sheath fluid has fully developed into a laminar flow by the time it reaches the outlet of the sample line SL, so that when the sample and sheath fluid are concentrated, they form a uniform, tight, and stable sample flow. In addition, the symmetrical arrangement of the fluid ports 120a and 120b also allows the sheath fluid to be injected symmetrically, improving fluid stability.
[0042] The base 100 may further include a defoaming channel 140. One end of the defoaming channel 140 communicates with channel 110, and the other end may be attached to a defoaming device such as a vacuum pump for completely releasing bubbles from channel 110. The defoaming channel 140 is positioned above channel 110 and higher than the fluid ports 120a and 120b to remove bubbles from the upper fluid ports 120a and 120b, thereby minimizing the impact of the defoaming process on fluid stability. At the connection between the defoaming channel 140 and channel 110, the end portion of the annular projection 210 of the cover member 200 is positioned as an inclined surface 212 to guide the release of bubbles and thus prevent bubbles from accumulating in the dead flow area above channel 110. In addition to removing bubbles, the defoaming channel 140 and the defoaming device can also function to clean up obstructions.
[0043] An annular cavity 150 surrounding the sample line SL is positioned above the base 100 to accommodate the annular piezoelectric element 60. The piezoelectric element 60 can be bonded within the annular cavity 150.
[0044] It should be understood that the structure of the base 100 and cover member 200 described above may be modified as needed and is not limited to the specific examples shown in Figures 1-2B. In other embodiments, the cover member 200 may be omitted.
[0045] The cuvette assembly 300 is a component that allows light to pass through for the optical detection of a sample, and is fixed below the exit 130 of the tapered section of the channel 110. The cuvette assembly 300 includes a light-transmitting cuvette body 310 having a sample detection channel 320. The sample detection channel 320 has an upper end aligned with the exit 130 of the tapered section and a lower end connected to the nozzle assembly 50 and aligned with the nozzle 510 in the nozzle assembly 50.
[0046] Figure 3 is an enlarged view of the cuvette assembly 300 in its assembled state. As shown in Figure 3, the cuvette assembly 300 is firmly fixed to a substantially plate-shaped positioning member 20 by a joint or other connection method that does not affect the optical performance of the cuvette assembly 300. During installation, high-precision assembly equipment is used to ensure that the sample detection channel 320 is concentrically aligned with the outlet 130 of the upstream channel 110. A tapered transition section 322 is positioned around the upper opening of the sample detection channel 320 to guide the liquid flow from the tapered section of the channel 110. The positioning member 20 is equipped with a limiting structure that receives and guides the nozzle 510 so that the nozzle 510 and the sample detection channel 320 are automatically aligned. Preferably, the nozzle 510 is independently removable without affecting its surrounding components, thereby allowing the nozzle 510 and the cuvette assembly 300 to be conveniently cleaned without moving the cuvette assembly 300, and eliminating the need to realign the detection light path after repeated disassembly and assembly of the cuvette assembly 300. In this embodiment, the nozzle assembly 50 further includes a carrier 520 for carrying the nozzle 510, which facilitates the independent removal of the nozzle 510. O-rings 920 and 930 for sealing are positioned around the joint between the sample detection channel 320 and the outlet 130 of channel 110, and around the joint between the sample detection channel 320 and the nozzle 510, respectively. The top of the cuvette assembly 300 is tightly pressed by a nut 940 to ensure positioning.
[0047] When a sample is optically inspected, it flows into channel 110 through an elongated sample line SL under pressure, is immediately encapsulated by a sheath fluid, and then flows into the cuvette assembly 300 along a tapered section of channel 110 with the sheath fluid. The sample is, for example, a single fluorescently labeled cell suspension. The sheath fluid surrounding the sample passes through a sample detection channel 320 in the cuvette body 310 and is discharged through a nozzle 510. A piezoelectric element 60 vibrates at a high frequency under the action of an electrical signal, thereby discharging from the nozzle 510, uniformly cutting the sheath fluid flow surrounding the sample droplet and forming separate droplets for subsequent sample sorting. In the sample detection channel 320, the sample droplet encapsulated by the sheath fluid is irradiated by a laser or another light source, emitting scattered light and fluorescence signals that reflect sample information. The optical system of the sample processor collects the optical signals, which are then processed and analyzed to detect and analyze the sample. Next, the sample sorting system sorts the samples discharged from the nozzle 510 based on the detection and analysis results.
[0048] The specific structure, operating principle, and beneficial effects of the cuvette assembly 300 will be described in detail below.
[0049] Figures 4A and 4B are three-dimensional views of the cuvette assembly 300 from different angles, Figure 4C is a top view of the cuvette assembly 300, and Figure 4D is a cross-sectional view obtained along the cross-sectional line CC in Figure 4C.
[0050] As shown in Figures 4A-4D, the cuvette body 310 within the cuvette assembly 300 is a substantially rectangular parallelepiped component, which can be made of fused silica glass or another suitable light-transmitting material. For ease of explanation, the long and short sides of the cuvette body 310 in a horizontal cross-section are defined. In this embodiment, the cuvette body 310 has a length of about 8.2 mm, a width of about 5.2 mm, and a height of about 10 mm. The sample detection channel 320 penetrates the cuvette body 310 perpendicularly in the height direction (z direction in Figures 4A-4D). As shown in Figure 4C, the sample detection channel 320 has a square cross-section.
[0051] The cuvette assembly 300 also includes a reflector 330, which is positioned on one side (hereinafter referred to as the first side) extending along one of the long sides of the cuvette body 310 and is used to reflect and focus the side-scattered and fluorescent light emitted by the sample droplet. The lower surface of the reflector 330 is coplanar with the lower surface of the cuvette body 310 so that the cuvette assembly 300 is mounted substantially on a plate-shaped positioning member 20. The reflector 330 is part of a spherical mirror and specifically includes a flat surface attached to the first side of the cuvette body 310 and a spherical surface opposite the flat surface, with the lower half cut off. The spherical surface is coated with a reflective film. The reflector 330 extends along the long side of the cuvette body 310 and beyond the short side of the cuvette body 310, i.e., the flat surface of the reflector 330 is slightly longer than the first side of the cuvette body 310. In this embodiment, the reflector 330 extends slightly beyond the front surface (hereinafter referred to as the second surface) of the cuvette body 310 along its short side, and the reflector 330 is coplanar with the back surface of the cuvette body 310. In this embodiment, the reflector 330 has a length of approximately 8.6 mm and a maximum height of approximately 5.4 mm.
[0052] The cuvette assembly 300 may further include a focusing lens 340 for shaping the incident light and focusing it into the sample detection channel 320. The focusing lens 340 may be mounted on a second side of the cuvette body 310 and adjacent to the reflector 330, slightly beyond the second side, so that the structure of the cuvette assembly 300 is smaller and light loss is reduced.
[0053] The cuvette assembly 300 may further include an aspherical lens 350 positioned opposite the reflector 330, the aspherical lens 350 shaping the focused spot of scattered and fluorescent light formed by the reflector 330 and focusing the spot into a signal detection device for detecting an optical signal. The aspherical lens 350 may be mounted on a third side of the cuvette body 310 opposite a first side so that the structure of the cuvette assembly 300 is smaller and optical loss is reduced. The aspherical lens 350 includes a shaping portion 352 for shaping and an outer frame 354 surrounding the shaping portion 352. The peripheral edge of the shaping portion 352 is substantially circular, and the central portion near the center of the shaping portion 352 is thicker than the peripheral portion.
[0054] Similar to the reflector 330, the lower surfaces of the focusing lens 340 and the aspherical lens 350 are coplanar with the lower surface of the cuvette body 310 to facilitate the installation of the cuvette assembly 300.
[0055] When a sample is optically detected, laser light is incident on the cuvette body 310 in the longitudinal direction (x direction in Figures 4A-4D) through the focusing lens 340. The focusing lens 340 shapes the incident laser light and focuses it into the sample detection channel 320. The focal point is called the laser inspection point P (shown in Figure 4D). When sample droplets (such as fluorescently labeled cells) enclosed in the sheath fluid are irradiated with laser light and pass through the laser inspection point P in the sample detection channel 320, scattered light and fluorescent light are emitted. In this embodiment, the laser inspection point P in the sample detection channel 320 is set at the center of the sphere on the spherical surface of the reflector 330 so that the reflector 330 can reflect and focus the scattered light and fluorescent light within its maximum range.
[0056] Figures 5A and 5B are schematic optical path diagrams of the side-scattered and fluorescent light being collected, as viewed from above and to the side of the cuvette assembly 300, respectively. As shown in Figures 5A and 5B, the scattered and fluorescent light diverging from the laser survey point P in various directions is reflected and focused by the reflector 330, then shaped by the aspherical lens 350 and focused onto the optical fiber OF of the signal detection device. The scattered and fluorescent light emitted by different laser-excited samples will be focused to different focal points after reflection, shaping, and focusing. For example, Figure 5B shows that different scattered or fluorescent light generated by the excitation of four different lasers can be distinguished.
[0057] The reflector 330 according to the present invention is not a perfectly spherical mirror, but a spherical mirror having a spherical surface with the lower half partially truncated. Therefore, the center of the sphere on the spherical surface of the reflector 330, i.e., the laser inspection point P, is closer to the nozzle 510, thereby reducing the droplet delay time from the laser inspection point P to the nozzle 510 and obtaining improved sorting performance. On the other hand, the area size of the flat surface of the reflector 330 affects the range that the reflector 330 can cover, which in turn affects the angular range of light that the reflector 330 can receive, so shortening the reflector will affect the efficiency of collecting side-scattered and fluorescent light. To compensate for the loss of light collection efficiency caused by a truncated reflector, the flat surface of the reflector 330 in the present invention is configured to be slightly longer than the first side of the cuvette body 310, thereby expanding the area of the reflector 330 that receives side-scattered and fluorescent light. The numerical aperture NA is commonly used in optical systems to measure the angular range of light that the system can collect. While the numerical aperture of conventional side-scattered and fluorescent light collection systems using lensing methods is approximately 0.6, the numerical aperture of the reflector 330 in the present invention, which can receive scattered light and fluorescence signals, can reach 1.25. Therefore, the cuvette assembly 300 according to the present invention improves the sorting performance of the sample processor, significantly improves the efficiency of collecting side-scattered and fluorescent light, and thus has higher resolution and sensitivity.
[0058] The cross-sectional shape and area of the sample detection channel 320 are important factors that affect the velocity of the sample droplet at the laser inspection point P. A faster velocity of the sample droplet passing through the laser inspection point P results in better sorting performance, but the increase in velocity will increase the difficulty of optical detection and weaken the optical performance. As shown in Figure 4C, in this embodiment, the cross-section of the sample detection channel 320 is square. The size of the sample detection channel 320 is determined based on factors such as the shape of the sample detection channel 320, the size of the sample droplet, the flow rates of the sample and sheath fluid, and the desired velocity of the sample droplet as it passes through the laser inspection point P. In this embodiment, the sample detection channel 320 is configured to have a square cross-section with a size of approximately 200 μm × 200 μm. The shape and size design of the sample detection channel 320 according to the present invention slightly increases the velocity of the sample droplet at the laser inspection point P compared to that of existing flow cytometers available on the market, so that sorting performance is improved without excessively reducing optical performance.
[0059] As can be seen more clearly from Figure 4C, the sample detection channel 320 is positioned approximately in the longitudinal direction of the cuvette body 310 so that the reflector 330 can capture and reflect scattered and fluorescent light within its maximum range. However, the sample detection channel 320 is positioned asymmetrically in the width direction (y direction in the drawing) of the cuvette body 310, that is, the distance between the sample detection channel 320 and the aspherical lens 350 is different from the distance between the sample detection channel 320 and the reflector 330. For example, in this embodiment, the distance from the center of the sample detection channel 320 to the reflector 330 is 2 mm, and the distance to the aspherical lens 350 is 3.2 mm. This asymmetrical design makes the cuvette body 310 thicker and increases the area of the cuvette body 310 adjacent to its upstream and downstream components, so that the high-pressure fluid can be sealed more tightly.
[0060] While the special design of the reflector 330 and the sample detection channel 320 makes the cuvette assembly 300 and flow cell 1 according to this disclosure more suitable for a sample sorter with sorting capabilities, it should be understood that such a cuvette assembly and flow cell may also be applicable to a sample analyzer or other sample processor without sorting capabilities.
[0061] While this disclosure is described with reference to exemplary embodiments, it should be understood that this disclosure is not limited to the specific embodiments described and illustrated herein. Those skilled in the art can make various modifications to the exemplary embodiments without departing from the scope defined by these claims. Features in the various embodiments can be combined with each other, provided they do not conflict. Alternatively, certain features in the embodiments may be omitted.
Claims
1. A cuvette assembly for a sample processor, wherein the cuvette assembly is A cuvette body having the shape of a rectangular parallelepiped, wherein the cuvette body is provided with a sample detection channel that penetrates the cuvette body perpendicularly, and the cuvette body has a long side and a short side in a horizontal cross-section, reflector and Equipped with, The reflector has a flat surface attached to a first side surface extending along one of the long sides of the cuvette body, and a spherical surface opposite the flat surface, with the lower half cut off. A cuvette assembly in which the reflector is positioned such that it is coplanar with the lower surface of the cuvette body, the center of the sphere on the spherical surface is located within the sample detection channel, the reflector extends along the long side, and the reflector extends beyond the short side.
2. The cuvette assembly according to claim 1, further comprising a focusing lens attached to a second side extending along one of the short sides of the cuvette body, wherein the focusing lens focuses incident light to the center of the sphere on the spherical surface of the reflector.
3. The cuvette assembly according to claim 2, further comprising an aspherical lens attached to a third side facing a first side of the cuvette body, wherein the aspherical lens shapes the focused spot formed by the reflection of the reflector so that the focused spot can be focused into a signal detection device for detecting an optical signal.
4. The cuvette assembly according to claim 3, wherein the aspherical lens comprises a molding portion for shaping and an outer frame surrounding the molding portion, the central portion of the molding portion being thicker than the peripheral portion.
5. The sample detection channel has a square cross-section, as described in any one of claims 1 to 4.
6. A flow cell for a sample processor, wherein the flow cell is Frame and, A flow cell body fixed to the frame, the flow cell body comprising a base and a cuvette assembly according to any one of claims 1 to 5, wherein the cuvette assembly is located below the base, and a sample from a sample line and a fluid from a fluid line are concentrated in the base and flow into the sample detection channel within the cuvette assembly. Nozzle assembly having a nozzle and Equipped with, The nozzle is located at the outlet of the sample detection channel and discharges the sample from the sample detection channel in a predetermined mode, forming a flow cell.
7. The flow cell according to claim 6, wherein the base comprises a vertical channel, the vertical channel having a smooth inner surface and comprising a smoothly transitioning cylindrical section and a tapered section, the tapered section being concentrically aligned with the sample detection channel, so that the sample and the fluid can be concentrated within the channel.
8. The flow cell according to claim 7, wherein the base further comprises symmetrically arranged fluid ports communicating with the fluid line and the vertical channel, the fluid ports being higher than the outlet of the sample line.
9. The flow cell according to claim 8, wherein the base further comprises a degassing channel, one end of the degassing channel communicating with the vertical channel and the other end being configured to be attached to a degassing device.
10. The flow cell according to claim 9, wherein the degassing channel is located above the vertical channel.
11. The flow cell according to claim 10, wherein the degassing channel is higher than the fluid port.
12. The flow cell according to claim 7, wherein the flow cell body further comprises a cover member, the cover member being located above the base.
13. The flow cell according to claim 12, wherein the flow cell body comprises an annular projection, the inner surface of which defines a sample port for enabling the sample line to be concentrically aligned with the vertical channel.
14. The flow cell according to any one of claims 12 or 13, wherein the end portion of the annular projection is configured to be inclined to induce the release of bubbles.
15. The flow cell according to claim 6, wherein the nozzle assembly is installed within the flow cell in a manner that allows it to be independently removed.
16. The flow cell according to claim 6, wherein the cavity is located on the upper part of the base and is configured to house a piezoelectric element.
17. The flow cell according to claim 16, wherein the cavity is an annular cavity surrounding the sample line.
18. A sample processor comprising a cuvette assembly according to any one of claims 1 to 5, or a flow cell according to any one of claims 6 to 17.
19. The sample processor according to claim 18, wherein the sample processor is a sample sorter.
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