CN112596250A - Illumination system of flow cytometry sorter and flow cytometry sorter - Google Patents
Illumination system of flow cytometry sorter and flow cytometry sorter Download PDFInfo
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- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
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Abstract
The invention discloses an illumination system of a flow cytometer and the flow cytometer, wherein the illumination system of the flow cytometer comprises a light source unit and a light beam shaping unit, and the light beam shaping unit is positioned on a transmission path of emergent light of the light source unit; the beam shaping unit comprises a cylindrical surface aspheric lens group, and the cylindrical surface aspheric lens group comprises a first conical surface and a second conical surface; the first conical surface has a first focal power
The second conical surface has a second focal power
Wherein
By adopting the technical scheme, the cylindrical aspheric lens group comprises two conical surfaces, and the focal power of the two conical surfaces is reasonably set, so that a flat-top elliptical light beam can be obtained after a light beam emitted by the light source unit passes through the light beam shaping unit, the light spot area of the flat-top elliptical light beam is larger, and the high cell or particle detection accuracy is ensured; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher.
Description
Technical Field
The embodiment of the invention relates to the technical field of cell sorting, in particular to an illumination system of a flow cytometry sorter and the flow cytometry sorter.
Background
Flow cytometry-based instruments, including flow cytometers, hematology analyzers, particle analyzers, and the like, are technical platforms for rapid quantitative analysis and sorting of cells or other microparticles arranged in a single column in a target flow one by one, and the basic principle is to irradiate a single cell or microparticle with a focused laser beam and analyze generated scattered light or fluorescence signals by using a photoelectric detector to obtain various parameters of an object to be detected. The laser light source and the optical system thereof are one of the core components of the flow cytometer, and the quality and stability of the focused light beam directly determine the performance index of the flow cytometer. Because the cell sample to be analyzed flows at high speed (5000/s) in the target flow, the time for passing through the laser irradiation area is only in the order of microseconds, and the intensity of the generated scattered light or fluorescence signal is closely related to the optical power density distribution of the laser irradiation area.
In order to ensure that each cell or particle passing through the focusing spot receives laser irradiation with the same intensity and ensure the analysis accuracy of the flow cytometer, the focusing spot size needs to be ensured to be large. However, in the prior art, the size of the focused light spot is increased either by increasing the spot size of the incident light beam or by adjusting the structure of the laser shaping device. However, increasing the spot size of the incident beam has high requirements on the laser, increasing the laser cost; the laser shaping device is difficult to process due to the fact that the structure of the laser shaping device is adjusted, flat-top portions of the shaped flat-top light beams are small in occupied ratio, and the laser energy utilization rate is low.
Disclosure of Invention
In view of this, embodiments of the present invention provide an illumination system for a flow cytometer and a flow cytometer, where the flow cytometer has a simple structure and a large size of a flat-top beam.
In a first aspect, an embodiment of the present invention provides an illumination system for a flow cytometer, including a light source unit and a beam shaping unit, where the beam shaping unit is located on a propagation path of an outgoing light of the light source unit;
the beam shaping unit comprises a cylindrical surface aspheric lens group, and the cylindrical surface aspheric lens group comprises a first conical surface and a second conical surface;
the first conical surface has a first focal power
The second conical surface has a second focal power
Wherein
Optionally, the radius of curvature of the first conical surface is Ry1The conic coefficient of the first conic surface is ky1And | Ry1|<100mm，-1000<ky1<-1；
The curvature radius of the second conical surface is Ry2The conic coefficient of the second conic surface is ky2And | Ry2|<100mm，-1000<ky2<-1。
Optionally, the focal length f1 of the first conical surface satisfies 0mm < f1<250mm, and the focal length f2 of the second conical surface satisfies-250 mm < f2<0 mm.
Optionally, the cylindrical aspheric lens group includes a first cylindrical aspheric lens and a second cylindrical aspheric lens, the first cylindrical aspheric lens includes the first conical surface, and the second cylindrical aspheric lens includes the second conical surface;
the first cylindrical aspheric lens is positioned on a propagation path of emergent light of the light source unit, and the second cylindrical aspheric lens is positioned on a propagation path of transmitted light obtained by transmission of the first cylindrical aspheric lens; or, the second cylindrical aspheric lens is located on a propagation path of the emergent light of the light source unit, and the first cylindrical aspheric lens is located on a propagation path of the transmitted light obtained by transmission through the second cylindrical aspheric lens.
Optionally, the cylindrical aspheric lens group includes a third cylindrical aspheric lens, and the third cylindrical aspheric lens includes the first conical surface and the second conical surface;
the first conical surface is positioned on a propagation path of emergent light of the light source unit, and the second conical surface is positioned on a propagation path of transmitted light obtained by transmission of the first conical surface; or, the second conical surface is located on a propagation path of the emergent light of the light source unit, and the first conical surface is located on a propagation path of the transmitted light obtained by transmission through the first conical surface.
Optionally, the emergent light of the light source unit passes through the beam shaping unit to obtain a flat-topped elliptical light beam;
along the long axis direction of the flat-topped elliptical beam, the light intensity of the flat-topped elliptical beam is uniformly distributed; along the short axis direction of the flat-topped elliptical beam, the light intensity of the flat-topped elliptical beam is in Gaussian distribution;
the spot size of the flat-topped elliptic light beam in the major axis direction is a first spot size, wherein the first spot size Wx is the light intensity Iw1Corresponding beam diameter, whereinw1Is 1/e of the maximum light intensity in the long axis direction2Doubling;
the second spot size Wt is the corresponding beam diameter when the light intensity uniformity meets a preset value, wherein the preset value a meets the conditions that a is more than or equal to 95% and less than or equal to 100%, and Wt/Wx is more than 75%.
Optionally, the light source unit includes a multi-wavelength light source unit.
Optionally, the multi-wavelength light source unit includes a plurality of single-mode fiber-coupled lasers;
the illumination system also comprises a plurality of collimating lenses and a plurality of light beam turning devices, wherein the collimating lenses are in one-to-one correspondence with the single-mode fiber coupled lasers, and the light beam turning devices are in one-to-one correspondence with the collimating lenses;
the collimating lens is positioned on a propagation path of emergent laser of the single-mode fiber coupled laser and is used for collimating the emergent laser to obtain a collimated laser beam;
the beam turning device is positioned on the transmission path of the collimated laser beam and used for reflecting the collimated laser beam to the geometric center of the cylindrical aspheric lens group.
Optionally, the illumination system further includes an achromatic double cemented lens, and the achromatic double cemented lens is located on a propagation path of the transmitted light obtained by transmission through the cylindrical aspheric lens group;
the focal length f3 of the achromatic doublet satisfies 20mm < f2<100 mm.
In a second aspect, embodiments of the present invention further provide a flow cytometer including the illumination system of the flow cytometer described in the first aspect.
The illumination system of the flow cytometry sorter comprises a light source unit and a light beam shaping unit, the light beam shaping unit comprises a cylindrical aspheric lens group, the cylindrical aspheric lens group comprises a first conical surface and a second conical surface, and the first conical surface is provided with a first focal power
The second conical surface has a second focal power
Wherein
So not only can guarantee cylindrical surface aspheric lens group simple structure in lighting system, guarantee cylindrical surface aspheric lens group and addThe process is simple; meanwhile, the illumination system provided by the embodiment of the invention can obtain the flat-top elliptical light beam, and the light spot area of the flat-top elliptical light beam is larger, so that the scattered light or fluorescence signal intensity generated by the cells or particles in the flow chamber under the irradiation of the larger flat-top elliptical light beam is ensured to be the same, and the detection accuracy of the cells or particles is ensured to be high; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher.
Drawings
Other features, objects and advantages of the invention will become more apparent upon reading of the detailed description of non-limiting embodiments made with reference to the following drawings:
fig. 1 is a schematic structural diagram of an illumination system of a flow cytometer according to an embodiment of the present invention;
FIG. 2 is a schematic cross-sectional view of an emergent light beam and a schematic light intensity distribution provided by an embodiment of the present invention;
FIG. 3 is a schematic cross-sectional view and intensity subdivision of a flattened elliptical beam provided by an embodiment of the present invention;
FIG. 4 is an enlarged schematic view of area A of FIG. 3;
FIG. 5 is a schematic diagram of an illumination system of another flow cytometer provided in accordance with an embodiment of the present invention;
FIG. 6 is a schematic diagram of an illumination system of another flow cytometer in accordance with an embodiment of the present invention;
FIG. 7 is a schematic diagram of an illumination system of another flow cytometer in accordance with an embodiment of the present invention;
FIG. 8 is a schematic diagram of an illumination system of another flow cytometer in accordance with an embodiment of the present invention;
FIG. 9 is a schematic diagram of a plurality of flattened elliptical beams according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be fully described by the detailed description with reference to the accompanying drawings in the embodiments of the present invention. It is obvious that the described embodiments are a part of the embodiments of the present invention, not all embodiments, and all other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without inventive efforts fall within the scope of the present invention.
Fig. 1 is a schematic structural diagram of an illumination system of a flow cytometer according to an embodiment of the present invention, and as shown in fig. 1, the illumination system of the flow cytometer according to the embodiment of the present invention includes a light source unit 10 and a beam shaping unit 20, where the beam shaping unit 20 is located on a propagation path of an outgoing light of the light source unit 10; the beam shaping unit 20 includes a cylindrical aspherical lens group 201, and the cylindrical aspherical lens group 201 includes a first conical surface 201a and a second conical surface 201 b; the first conical surface 201a has a first refractive power
The second conical surface 201b has the second refractive power
Wherein
Illustratively, as shown in fig. 1, the outgoing light beam 101 of the light source unit 10 passes through the beam shaping unit 20 and then is irradiated onto the flow cell 40, and is converged at the central position of the flow cell 40. The emergent light beam 101 may be a collimated gaussian spot, and its cross-sectional view and light intensity distribution are shown in fig. 2. Wherein I1Is the intensity at the center of the spot, i.e. the maximum intensity of the outgoing light beam 101, I2Is I11/e of2And (4) doubling. W1As a light intensity of I2Corresponding beam diameter. In the present invention, W1The thickness may be any value between 0.5mm and 5mm, which is not limited in the embodiment of the present invention. Further, the light source unit 10 may be a single-wavelength light source unit or a multi-wavelength light source unit, which is not limited in the embodiment of the present invention.
The cylindrical aspherical lens group 201 includes a first conical surface 201a and a second conical surfaceA surface 201b, wherein the first conical surface 201a has positive power, and the second conical surface 201b has negative power, i.e. the first power of the first conical surface 201a
Satisfy the requirement of
Second focal power of the second conical surface 201b
Satisfy the requirement of
Therefore, the emitted laser 101 can be ensured to be a flat-top elliptical beam after passing through the cylindrical aspheric lens group 201, and the flat-top elliptical beam has a larger light spot, so that the scattered light or fluorescence signal intensity generated by the cells or particles in the flow chamber 40 under the irradiation of the larger flat-top elliptical beam is ensured to be the same, and the detection accuracy of the cells or particles is ensured to be high.
Optionally, the cylindrical aspheric lens group 201 may include one or more cylindrical aspheric lenses, which is not limited in the embodiment of the present invention, and it is only necessary to ensure that the cylindrical aspheric lens group 201 includes two conical surfaces with opposite focal powers. Further, the cylindrical aspheric lens may be a powell prism or other shaping lens, which is not limited in the embodiment of the present invention.
In summary, in the illumination system for flow cytometry sorting provided in the embodiment of the present invention, the light beam shaping unit is provided with two conical surfaces, and the focal powers of the two conical surfaces are opposite, so that it is ensured that the light spot of the flat-top elliptical light beam obtained by the light beam shaping unit is relatively large, the scattered light or fluorescence signal intensity generated by the cells or particles in the flow chamber under the irradiation of the relatively large flat-top elliptical light beam is the same, and the detection accuracy of the cells or particles is high; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher; in addition, the illumination system provided by the embodiment of the invention has a simple structure, can ensure that the flat-topped elliptical beam with a large light spot can be obtained only by arranging the beam shaping unit to comprise two conical surfaces with opposite focal powers, has a simple preparation process, improves the preparation yield of the beam shaping unit, and reduces the preparation cost of the beam shaping unit.
Specifically, fig. 3 is a schematic cross-sectional view and a schematic light intensity distribution view of a flat-topped elliptical light beam provided by an embodiment of the present invention, and fig. 4 is an enlarged schematic view of an area a in fig. 3, as shown in fig. 3 and 4, an emergent light ray 101 of a light source unit 10 passes through a light beam shaping unit 20 to obtain the flat-topped elliptical light beam; along the long axis direction of the flat-topped elliptical beam, such as the X direction shown in the figure, the light intensity of the flat-topped elliptical beam is uniformly distributed; along the short axis direction of the flat-topped elliptical beam, such as the Y direction shown in the figure, the light intensity of the flat-topped elliptical beam is Gaussian distributed; the spot size of the flat-topped elliptic light beam in the major axis direction is a first spot size, wherein the first spot size WxIs the light intensity Iw1Corresponding beam diameter, whereinw1Is 1/e of the maximum light intensity in the long axis direction2Doubling; the second spot size Wt is the corresponding beam diameter when the light intensity uniformity meets a preset value, wherein the preset value a meets the conditions that a is more than or equal to 95% and less than or equal to 100%, and Wt/Wx>75 percent; wherein, the calculation formula of the light intensity uniformity U is as follows:
Imaxand IminRespectively representing the maximum light intensity value and the minimum light intensity value of the flat-topped elliptic light beam in the major axis direction.
For example, as shown in fig. 3 and 4, the outgoing light beam 101 passes through the beam shaping unit 20 to obtain a flat-top elliptical light beam, which is uniformly distributed in the major axis direction, i.e., the X direction shown in the figure, so as to ensure that the intensity of the scattered light or fluorescence signal generated by the cells or particles in the flow cell under the irradiation of the larger flat-top elliptical light beam is the same, and ensure high cell or particle detection accuracy.
Further, in the embodiment of the present invention, the second light spot size Wt is a light beam diameter corresponding to a light intensity uniformity satisfying a preset value, where a may satisfy 95% to 100%, that is, a ratio of the light beam diameter Wt corresponding to a uniformity greater than 95% to the light beam diameter Wx of the flat-top elliptical light beam in the major axis direction is greater than 75%, as shown in fig. 3 and 4, it is fully ensured that the flat-top elliptical light beam has good uniformity in the major axis direction, it is ensured that the scattered light or fluorescent signal intensities generated by the cells or particles in the flow chamber under the irradiation of the larger flat-top elliptical light beam are the same, and it is ensured that the detection accuracy of the cells or particles is high. Furthermore, the preset value in the embodiment of the invention can even reach 98%, and the flat-topped elliptical beam is fully ensured to have good uniformity in the major axis direction.
For example, in the embodiment of the present invention, the beam diameter Wx of the flattened elliptical beam in the major axis direction may be 60 μm, and Wt may be greater than 45 μm. Furthermore, the beam diameter Wy of the flat-topped elliptical beam in the short axis direction can be 6-13 μm. Further, the corresponding beam diameter Wt when the uniformity meets the preset value may be any value between 20 μm and 60 μm, which is not limited in the embodiment of the present invention, and it is only exemplified that Wt may be greater than 45 μm.
Optionally, the surface type function of the first conical surface 201a may satisfy:
wherein the content of the first and second substances,
Ry1is the radius of curvature, k, of the first conical surface 201ay1Is the conic coefficient of the first conic surface 201a, and | Ry1|<100mm，-1000<ky1<-1；
The surface type function of the second conical surface 201b may satisfy:
wherein the content of the first and second substances,
Ry2is the radius of curvature, k, of the second conical surface 201by2Is the conic coefficient of the second conic surface 201b, and | Ry2|<100mm，-1000<ky2<-1。
It can be understood that, according to the surface type function of the first conical surface 201a and the surface type function of the second conical surface 201b, the surface type functions of the first conical surface 201a and the second conical surface 201b are only related to variables in a single direction (y direction), so that the surface type functions of the first conical surface 201a and the second conical surface 201b are simple, the first conical surface 201a and the second conical surface 201b are simple in structure, the manufacturing process is simple, and high manufacturing efficiency and manufacturing yield of the first conical surface 201a and the second conical surface 201b can be ensured.
It should be noted that the above embodiment is described by taking only one possible surface function of the first conical surface 201a and the second conical surface 201b as an example, and it is understood that the surface function of the first conical surface 201a may also satisfy other functional expressions, such as:
wherein at least one of a1, a2, a3, a4, a5, a6, a7, and a8 is non-zero.
The surface-type function of the second conical surface 201b may also satisfy other functional expressions, such as:
wherein at least one of a1 ', a 2', a3 ', a 4', a5 ', a 6', a7 'and a 8' is non-zero.
In this way, the surface functions of the first conical surface 201a and the second conical surface 201b are also only related to variables in a single direction (y direction), so that the surface functions of the first conical surface 201a and the second conical surface 201b are simple, the first conical surface 201a and the second conical surface 201b can be ensured to have simple structures, the preparation process is simple, and the preparation efficiency and the preparation yield of the first conical surface 201a and the second conical surface 201b can be ensured to be high. Further, the curvature radius and cone coefficient of the first conical surface 201a and the curvature radius and cone coefficient of the second conical surface 201b are reasonably set, so that the light spot of the flat-topped elliptical beam obtained by the beam shaping unit 20 can be ensured to be larger, the detection accuracy of cells or particles is ensured to be high, and the energy utilization rate of the emergent laser is ensured to be higher.
Alternatively, the focal length f1 of the first conical surface 201a may satisfy 0mm < f1<250mm, and the focal length f2 of the second conical surface 201b may satisfy-250 mm < f2<0 mm. The focal length of the first conical surface 201a and the focal length of the second conical surface 201b are reasonably set, so that the light spot of the flat-topped elliptical beam obtained by the beam shaping unit 20 can be ensured to be larger, and the detection accuracy of cells or particles and the energy utilization rate of the emergent laser are ensured to be higher.
Alternatively, the cylindrical aspheric lens group 201 may include one or more cylindrical aspheric lenses, and the first conical surface 201a and the second conical surface 201b may be respectively disposed on the same cylindrical aspheric lens or on different cylindrical aspheric lenses, which will be described in detail below for different cases.
First, the following description will be given taking an example in which the cylindrical aspherical lens group 201 includes two cylindrical aspherical lenses.
Optionally, with continued reference to fig. 1, the cylindrical aspheric lens group 201 includes a first cylindrical aspheric lens 2011 and a second cylindrical aspheric lens 2012, the first cylindrical aspheric lens 2011 includes a first conical surface 201a, and the second cylindrical aspheric lens 2012 includes a second conical surface 201 b; the first cylindrical aspheric lens 2011 is located on a propagation path of the outgoing light 101 from the light source unit 10, and the second cylindrical aspheric lens 2012 is located on a propagation path of the transmitted light transmitted through the first cylindrical aspheric lens 2011. Thus, the emergent light beam 101 sequentially passes through the first cylindrical aspheric lens 2011 and the second cylindrical aspheric lens 2012 to obtain a flat-top elliptical light beam, and the light spot of the flat-top elliptical light beam is large, so that the scattered light or fluorescence signal intensity generated by cells or particles in the flow chamber under the irradiation of the large flat-top elliptical light beam is ensured to be the same, and the detection accuracy of the cells or particles is ensured to be high; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher.
Optionally, fig. 5 is a schematic structural diagram of an illumination system of another flow cytometer according to an embodiment of the present invention, and as shown in fig. 5, the cylindrical aspheric lens group 201 includes a first cylindrical aspheric lens 2011 and a second cylindrical aspheric lens 2012, the first cylindrical aspheric lens 2011 includes a first conical surface 201a, and the second cylindrical aspheric lens 2012 includes a second conical surface 201 b; the second cylindrical aspheric lens 2012 is located on a propagation path of the outgoing light of the light source unit 10, and the first cylindrical aspheric lens 2011 is located on a propagation path of the transmitted light transmitted through the second cylindrical aspheric lens 2012. Thus, the emergent light beam 101 sequentially passes through the second cylindrical aspheric lens 2012 and the first cylindrical aspheric lens 2011 to obtain a flat-top elliptical light beam, and the light spot of the flat-top elliptical light beam is large, so that the scattered light or fluorescence signal intensity generated by cells or particles in the flow chamber under the irradiation of the large flat-top elliptical light beam is ensured to be the same, and the detection accuracy of the cells or particles is ensured to be high; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher.
The above description is made on the case where the lenticular aspherical lens group 201 includes two lenticular aspherical lenses, and next, the description is made on the case where the lenticular aspherical lens group 201 includes one lenticular aspherical lens.
Optionally, fig. 6 is a schematic structural diagram of an illumination system of another flow cytometer according to an embodiment of the present invention, as shown in fig. 6, the cylindrical aspheric lens group 201 includes a third cylindrical aspheric lens 2013, and the third cylindrical aspheric lens 2013 includes a first conical surface 201a and a second conical surface 201 b; the first conical surface 201a is located on the propagation path of the outgoing light of the light source unit 10, and the second conical surface 201b is located on the propagation path of the transmitted light transmitted through the first conical surface 201 a. Thus, the emergent light beam 101 sequentially passes through the first conical surface 201a and the second conical surface 201b to obtain a flat-top elliptical light beam, and the light spot of the flat-top elliptical light beam is large, so that the scattered light or fluorescent signal intensity generated by cells or particles in the flow chamber under the irradiation of the large flat-top elliptical light beam is ensured to be the same, and the detection accuracy of the cells or particles is ensured to be high; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher.
Optionally, fig. 7 is a schematic structural diagram of an illumination system of another flow cytometer according to an embodiment of the present invention, as shown in the drawing, the cylindrical aspheric lens group 201 includes a third cylindrical aspheric lens 2013, and the third cylindrical aspheric lens 2013 includes a first conical surface 201a and a second conical surface 201 b; the second conical surface 201b is located on the propagation path of the outgoing light of the light source unit 10, and the first conical surface 201a is located on the propagation path of the transmitted light transmitted through the second conical surface 201 b. Thus, the emergent light beam 101 sequentially passes through the second conical surface 201b and the first conical surface 201a to obtain a flat-top elliptical light beam, and the light spot of the flat-top elliptical light beam is large, so that the scattered light or fluorescent signal intensity generated by cells or particles in the flow chamber under the irradiation of the large flat-top elliptical light beam is ensured to be the same, and the detection accuracy of the cells or particles is ensured to be high; meanwhile, the energy utilization rate of the emergent laser can be ensured to be higher.
The foregoing embodiment has described in detail different composition manners of the cylindrical aspheric lens group 201, and it is understood that, in the embodiment of the present invention, the specific composition manner of the cylindrical aspheric lens group 201 is not limited, and the cylindrical aspheric lens group 201 may include two cylindrical aspheric lenses, as shown in fig. 1 and fig. 5, so that it can be ensured that each cylindrical aspheric lens has a simple structure and a simple preparation process; alternatively, the cylindrical aspheric lens group 201 may also include a cylindrical aspheric lens, as shown in fig. 6 and 7, so as to ensure that the cylindrical aspheric lens group 201 has a simple and compact structure and high integration. Further, in the embodiment of the present invention, the outgoing light beam 101 may first pass through the first conical surface 201a and then pass through the second conical surface 201b, as shown in fig. 1 and fig. 6; alternatively, the outgoing light beam 101 may also pass through the second conical surface 201b first and then pass through the first conical surface 201a, as shown in fig. 5 and 7, which is not limited in the embodiment of the present invention, and it is only required to ensure that the outgoing light beam 101 may pass through the first conical surface 201a and the second conical surface 201b, and be shaped by the first conical surface 201a and the second conical surface 201b to obtain a flattened elliptical light beam with a large light spot.
Further, the light source unit 10 may include a multi-wavelength light source unit 11, which ensures that the wavelength range of the light source unit 10 is large, for example, the entire visible light range is covered, and ensures that the entire illumination system can obtain flat top lights with different wavelength ranges, and is suitable for sorting different types of cells.
Further, fig. 8 is a schematic structural diagram of another illumination system of a flow cytometer provided in the embodiment of the present invention, and as shown in fig. 8, the multi-wavelength light source unit 11 includes a plurality of single-mode fiber coupled lasers 102, the illumination system of the flow cytometer provided in the embodiment of the present invention may further include a plurality of collimating lenses 50 and a plurality of beam bending devices 60, the collimating lenses 50 correspond to the single-mode fiber coupled lasers 102 one to one, and the beam bending devices 60 correspond to the collimating lenses 50 one to one; the collimating lens 50 is located on a propagation path of the emergent laser of the single-mode fiber coupled laser 102, and is configured to collimate the emergent laser to obtain a collimated laser beam; the beam-bending device 60 is located on the propagation path of the collimated laser beam 50, and is used for reflecting the collimated laser beam to the geometric center of the cylindrical aspheric lens group 201.
It can be understood that the illumination system of the flow cytometer provided by the embodiment of the present invention can be applied to a plurality of different laser light sources. Illustratively, as shown in fig. 8, the plurality of single-mode fiber-coupled lasers 102 may be 102a, 102b, and 102c, respectively, the plurality of collimating lenses 50 may be 50a, 50b, and 50c, respectively, and the plurality of beam-bending devices 60 may be 60a, 60b, and 60c, respectively. Wherein, the collimating lens 50a corresponds to the single mode fiber coupled laser 102a, and the beam steering device 60a corresponds to the collimating lens 50 a; the collimating lens 50b corresponds to the single-mode fiber coupled laser 102b, and the beam steering device 60b corresponds to the collimating lens 50 b; the collimating lens 50c corresponds to the single mode fiber coupled laser 102c, and the beam steering device 60c corresponds to the collimating lens 50 c.
Specifically, the collimator lenses 50a, 50b, and 50c collimate the divergent laser beam emitted from the single-mode fiber-coupled laser 102 into a beam of a specific aperture having a smaller divergence angle. The collimating lens 50 may be a fixed focus lens or a zoom lens, which is not limited in this embodiment of the present invention, and the light beam emitted after being collimated by the collimating lens 50 is still a gaussian light beam, and the quality factor of the light beam is less than 1.2.
Further, the beam-bending devices 60a, 60b and 60c are used to combine the collimated laser beams with different wavelengths, and then the combined laser beams are incident into the beam shaping unit 20 at a specific angle and position, and finally are incident into the central position of the flow cell 40. As shown in fig. 8, the beam-bending devices 60a, 60b and 60c are used to integrate the collimated laser beams with different wavelengths into the same position of the beam shaping unit 20, so that the positions and deflection angles of the beam-bending devices 60 need to be set reasonably to ensure that the laser beams with different wavelengths can be integrated after passing through the beam-bending devices 60.
Furthermore, the diameter d of the fiber core of the single-mode fiber coupled laser 102 can satisfy 0< d <5 μm, and the laser wavelength λ emitted by the single-mode fiber coupled laser 102 can satisfy 370nm ≤ λ ≤ 700 nm.
Illustratively, the diameter d of the fiber core of the single-mode fiber coupled laser 102 is set to satisfy 0< d <5 μm, so that the diameter of the fiber core of the single-mode fiber coupled laser 102 is ensured to be small, and the quality of the emergent light beam is ensured to be good enough. Furthermore, the laser wavelength λ emitted by the single-mode fiber coupled laser 102 meets the condition that λ is more than or equal to 370nm and less than or equal to 700nm, and the illumination system provided by the embodiment of the invention is ensured to be suitable for the whole visible light range.
Further, fig. 9 is a schematic diagram of a plurality of flat-topped elliptical beams according to an embodiment of the present invention, as shown in fig. 9, laser beams with different wavelengths are incident on the beam shaping unit 20 at the same position of the beam shaping unit 20, and are separately incident on different positions of the flow chamber 40 after passing through the beam shaping unit 20. The light spots corresponding to the three laser beams can be arranged at equal intervals, and the distance d between any two adjacent light spots satisfies that d is more than or equal to 120 mu m and less than or equal to 250 mu m.
Further, when the light source unit 10 may include a multi-wavelength light source unit 11 corresponding to a plurality of laser beams with different wavelengths, the illumination system of the flow cytometer provided in the embodiment of the present invention may further include an achromatic lens to eliminate the chromatic aberration problem caused by the laser beams with different wavelengths. Referring to fig. 1, 5, 6, 7 and 8, the illumination system of the flow cytometer provided by the embodiment of the present invention may further include an achromatic doublet 30, where the achromatic doublet 30 is located on the propagation path of the transmitted light obtained by transmitting through the cylindrical aspheric lens group 201; the focal length f3 of achromatic doublet 30 may satisfy 20mm < f2<100 mm.
Illustratively, in order to eliminate the problem of chromatic aberration caused by laser beams with different wavelengths, the illumination system of the flow cytometer provided in the embodiment of the present invention may further include an achromatic lens, and the embodiment of the present invention is only described by taking an achromatic cemented doublet 30 as an example, so that the chromatic aberration caused by the laser beams with different wavelengths is eliminated, and at the same time, the achromatic lens may be ensured to have a simple structure. Further, the focal length f3 of the achromatic double-cemented lens 30 can meet the requirement that the focal length f2 is less than 20mm and less than 100mm, the focal length of the achromatic double-cemented lens 30 is reasonably set, the achromatic double-cemented lens 30 can be matched with the size of an incident beam, and the size of a finally obtained flat-top light spot is ensured to meet the requirement.
Based on the same inventive concept, an embodiment of the present invention further provides a flow cytometer, including an illumination system of the flow cytometer according to the embodiment of the present invention, where the flow cytometer and the illumination system of the flow cytometer have the same beneficial effects, and are not described herein again.
It is to be noted that the foregoing is only illustrative of the preferred embodiments of the present invention and the technical principles employed. Those skilled in the art will appreciate that the present invention is not limited to the specific embodiments described herein, and that the features of the various embodiments of the invention may be partially or fully coupled to each other or combined and may be capable of cooperating with each other in various ways and of being technically driven. Numerous variations, rearrangements, combinations, and substitutions will now become apparent to those skilled in the art without departing from the scope of the invention. Therefore, although the present invention has been described in greater detail by the above embodiments, the present invention is not limited to the above embodiments, and may include other equivalent embodiments without departing from the spirit of the present invention, and the scope of the present invention is determined by the scope of the appended claims.
Claims (10)
1. An illumination system of a flow cytometer comprises a light source unit and a beam shaping unit, wherein the beam shaping unit is positioned on a propagation path of emergent light rays of the light source unit;
the beam shaping unit comprises a cylindrical surface aspheric lens group, and the cylindrical surface aspheric lens group comprises a first conical surface and a second conical surface;
the first conical surface has a first focal power
The second conical surface has a second focal power
Wherein
2. The illumination system of claim 1, wherein the first conical surface has a radius of curvature Ry1The conic coefficient of the first conic surface is ky1And | Ry1|<100mm，-1000<ky1<-1；
The curvature radius of the second conical surface is Ry2The conic coefficient of the second conic surface is ky2And | Ry2|<100mm，-1000<ky2<-1。
3. The illumination system of claim 1, wherein the focal length f1 of the first conical surface satisfies 0mm < f1<250mm, and the focal length f2 of the second conical surface satisfies-250 mm < f2<0 mm.
4. The illumination system of any of claims 1-3, wherein the cylindrical aspheric lens group includes a first cylindrical aspheric lens and a second cylindrical aspheric lens, the first cylindrical aspheric lens including the first conical surface and the second cylindrical aspheric lens including the second conical surface;
the first cylindrical aspheric lens is positioned on a propagation path of emergent light of the light source unit, and the second cylindrical aspheric lens is positioned on a propagation path of transmitted light obtained by transmission of the first cylindrical aspheric lens; or, the second cylindrical aspheric lens is located on a propagation path of the emergent light of the light source unit, and the first cylindrical aspheric lens is located on a propagation path of the transmitted light obtained by transmission through the second cylindrical aspheric lens.
5. The illumination system of any of claims 1-3, wherein the cylindrical aspheric lens group includes a third cylindrical aspheric lens that includes the first conical surface and the second conical surface;
the first conical surface is positioned on a propagation path of emergent light of the light source unit, and the second conical surface is positioned on a propagation path of transmitted light obtained by transmission of the first conical surface; or, the second conical surface is located on a propagation path of the emergent light of the light source unit, and the first conical surface is located on a propagation path of the transmitted light obtained by transmission through the first conical surface.
6. The illumination system of claim 1, wherein the outgoing light of the light source unit passes through the beam shaping unit to obtain a flat-topped elliptical beam;
along the long axis direction of the flat-topped elliptical beam, the light intensity of the flat-topped elliptical beam is uniformly distributed; along the short axis direction of the flat-topped elliptical beam, the light intensity of the flat-topped elliptical beam is in Gaussian distribution;
the spot size of the flat-topped elliptic light beam in the major axis direction is the first lightSpot size, wherein the first spot size Wx is the intensity of light Iw1Corresponding beam diameter, whereinw1Is 1/e of the maximum light intensity in the long axis direction2Doubling;
the second spot size Wt is the corresponding beam diameter when the light intensity uniformity meets a preset value, wherein the preset value a meets the conditions that a is more than or equal to 95% and less than or equal to 100%, and Wt/Wx is more than 75%.
7. The illumination system of claim 1, wherein the light source unit comprises a multi-wavelength light source unit.
8. The illumination system of claim 7, wherein the multi-wavelength light source unit comprises a plurality of single-mode fiber-coupled lasers;
the illumination system also comprises a plurality of collimating lenses and a plurality of light beam turning devices, wherein the collimating lenses are in one-to-one correspondence with the single-mode fiber coupled lasers, and the light beam turning devices are in one-to-one correspondence with the collimating lenses;
the collimating lens is positioned on a propagation path of emergent laser of the single-mode fiber coupled laser and is used for collimating the emergent laser to obtain a collimated laser beam;
the beam turning device is positioned on the transmission path of the collimated laser beam and used for reflecting the collimated laser beam to the geometric center of the cylindrical aspheric lens group.
9. The illumination system of claim 8, further comprising an achromatic doublet on a propagation path of transmitted light transmitted through the cylindrical aspheric lens group;
the focal length f3 of the achromatic doublet satisfies 20mm < f2<100 mm.
10. A flow cytometric analyzer, comprising an illumination system of the flow cytometric analyzer of any of claims 1 to 9.
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| WO2023129548A1 (en) * | 2021-12-30 | 2023-07-06 | Illumina, Inc. | Imaging systems and related methods |
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