CN114112873A - Waveguide design for on-chip fluorescence dispersive optical path for flow cytometer - Google Patents
Waveguide design for on-chip fluorescence dispersive optical path for flow cytometer Download PDFInfo
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- CN114112873A CN114112873A CN202111393323.8A CN202111393323A CN114112873A CN 114112873 A CN114112873 A CN 114112873A CN 202111393323 A CN202111393323 A CN 202111393323A CN 114112873 A CN114112873 A CN 114112873A
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Abstract
本发明公开了一种用于流式细胞仪的片上荧光色散光路的波导设计，包括棱镜组和波导阵列，所述棱镜组在流式细胞仪的片上荧光色散光路中用于将细胞发出的原始荧光进行色散分离，以输出各波长已经分离的荧光；所述波导阵列布置在所述各波长已经分离的荧光的后方，将所述各波长已经分离的荧光中的不同波长的光导入所述波导阵列的不同波导中，通过所述波导阵列中的不同波导的弯曲走向使不同波长的光在空间上进一步分离，在小尺寸的范围内实现更高的线色散率，满足PMT或其他光电检测器件的适配要求。本发明在小尺寸下依然实现足够的色散率，利于流式细胞仪的微型化和小型化设计。
The invention discloses a waveguide design for an on-chip fluorescence dispersion optical path of a flow cytometer, comprising a prism group and a waveguide array. The prism group is used in the on-chip fluorescence dispersion optical path of the flow cytometer to transmit the original The fluorescence is dispersed and separated to output the fluorescence whose wavelengths have been separated; the waveguide array is arranged behind the fluorescence whose wavelengths have been separated, and light of different wavelengths in the fluorescence whose wavelengths have been separated is guided into the waveguide In the different waveguides of the array, the light of different wavelengths can be further separated in space by the bending direction of the different waveguides in the waveguide array, and a higher linear dispersion rate can be realized in the range of small size, which can meet the requirements of PMT or other photoelectric detection devices. adaptation requirements. The invention still achieves sufficient dispersion rate under small size, which is beneficial to the miniaturization and miniaturization design of the flow cytometer.
Description
Technical Field
The invention relates to the technical field of flow cytometry, in particular to a waveguide design of an on-chip fluorescence dispersion optical path for a flow cytometer.
Background
Flow cytometry is an instrument that can rapidly detect biophysical and biochemical information of each cell or biological particle in a cell population, and is widely used in scientific research, clinical testing, and production activities. Fluorescence detection is the main means for cell detection by flow cytometry at present. The traditional flow cytometer uses a multi-channel light filtering method for fluorescence detection, and the method for directly detecting full-spectrum fluorescence is a novel fluorescence detection method. When detecting full-spectrum fluorescence, since the light intensity of the fluorescence signal of each wavelength is very weak, high-performance photoelectric detection devices, such as PMT and APD, are required to convert the optical signal of the fluorescence of each wavelength into an electrical signal for detection. High performance photodetectors tend to have large spatial dimensions, and when multiple photodetectors are used in parallel, the spatial distance between the fluorescent channels of each wavelength must be large to direct the fluorescent light of different wavelengths into different photomultiplier tubes. The prism system has a limited angular dispersion, and in order to achieve a sufficient linear dispersion, the distance between the prism and the photomultiplier in the prism system must be large, which in turn increases the spatial size of the flow cytometer.
In view of the specific requirements of flow cytometers, especially the demand for miniaturization of flow cytometers that has recently emerged, prism dispersion is the dominant dispersion method applicable to miniature flow cytometers. Miniaturization in turn requires that the dispersion function be implemented in a small-sized space, such as the size of a chip. In this context, prism dispersion cannot avoid the problem of insufficient dispersion ratio.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, an object of the present invention is to provide a waveguide design for an on-chip fluorescence dispersion optical path of a flow cytometer, which can achieve a sufficient dispersion ratio even in a small size, and is favorable for miniaturization and miniaturization design of the flow cytometer.
A waveguide design for an on-chip fluorescence-dispersive optical path for a flow cytometer according to an embodiment of the present invention comprises:
the prism group is used for carrying out dispersion separation on original fluorescence emitted by cells in an on-chip fluorescence dispersion optical path of the flow cytometer so as to output the fluorescence with separated wavelengths;
the waveguide array is arranged behind the fluorescence with the separated wavelengths, the light with different wavelengths in the fluorescence with the separated wavelengths is guided into different waveguides of the waveguide array, the light with different wavelengths is further separated in space through the bending trend of the different waveguides in the waveguide array, the higher linear dispersion rate is realized in a small-size range, and the adaptation requirement of a PMT or other photoelectric detection devices is met.
The waveguide design of the on-chip fluorescence dispersion optical path for the flow cytometer has the following advantages: the introduction of the array waveguide can ensure that enough dispersion rate can be still realized under small size, the fluorescence detection function of the spectral flow cytometer can be completed on a chip, the volume size of the flow cytometer can be effectively reduced, the integration of a flow cytometer system is facilitated, and the requirements of miniaturization and microminiaturization of the flow cytometer system are met; the light path stability is strong, only need general horizontally mechanical location and fixed can work, need not frequent calibration in the use.
According to some embodiments of the present invention, the waveguide design for on-chip fluorescence dispersion optical path of flow cytometer further comprises a first convex lens disposed behind the optical path of the prism group and near the last prism in the prism group for converging light of different wavelengths in the fluorescence light whose each wavelength has been separated at the position of different waveguide entrance of the waveguide array.
According to some embodiments of the present invention, the prism assembly includes a plurality of triangular prisms closely arranged in an arc shape.
According to some embodiments of the invention, the exit direction of the fluorescence light whose wavelengths have been separated is at most totally deflected by 180 degrees with respect to the incident direction of the original fluorescence light.
According to some embodiments of the present invention, different waveguide inlets in the waveguide array are closely arranged in sequence, and different waveguide outlets in the waveguide array are distributed in sequence, so as to achieve a higher linear dispersion ratio in a small size range, and meet the adaptation requirement of the PMT.
According to some embodiments of the present invention, the waveguide design for an on-chip fluorescence-dispersive optical path of a flow cytometer further comprises a second convex lens or lens group, arranged in a one-to-one correspondence behind the different waveguide outlets of the waveguide array, that focuses light exiting the waveguide for subsequent detection of fluorescence by the PMT.
According to some embodiments of the present invention, the waveguide design for the on-chip fluorescence dispersion optical path of the flow cytometer employs processing a mold on a silicon wafer and processing a waveguide structured by an optical dispersion material on a wafer through one-step molding by a reverse mold, or through microelectronic and MEMS processing technologies.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
fig. 1 is a schematic structural diagram of a waveguide design for an on-chip fluorescence-dispersive optical path of a flow cytometer according to an embodiment of the present invention.
Fig. 2 is a schematic structural diagram of a waveguide array in a waveguide design of an on-chip fluorescence-dispersive optical path for a flow cytometer according to an embodiment of the present invention.
Fig. 3 is an enlarged schematic view of the waveguide inlet of the waveguide array in the waveguide design of the on-chip fluorescence dispersion optical path for the flow cytometer according to the embodiment of the present invention.
Fig. 4 is an enlarged schematic diagram of a waveguide outlet of a waveguide array in a waveguide design of an on-chip fluorescence dispersive optical path for a flow cytometer according to an embodiment of the present invention.
Fig. 5 is an enlarged schematic view of a second convex lens at the waveguide exit of the waveguide array in the waveguide design of the on-chip fluorescence-dispersive optical path for a flow cytometer according to an embodiment of the present invention.
Fig. 6 is a simulation verification experiment result of the waveguide design of the on-chip fluorescence dispersion optical path for the flow cytometer according to the embodiment of the present invention.
FIG. 7 is a diagram of a processing tool for designing a waveguide on a silicon chip for an on-chip fluorescence-dispersive optical path of a flow cytometer according to an embodiment of the present invention.
Fig. 8 is a schematic diagram of the processing of a waveguide design based on optically dispersive materials for an on-chip fluorescence dispersive optical path of a flow cytometer in accordance with an embodiment of the present invention.
Reference numerals:
waveguide design 1000 for on-chip fluorescence dispersive optical path for flow cytometry
Waveguide array 2 waveguide 201 waveguide inlet 2011 waveguide outlet 2012
First convex lens 3, second convex lens 4, wafer substrate 5, reflecting layer 6
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
Waveguide design 1000 for an on-chip fluorescence-dispersive optical path for a flow cytometer in accordance with an embodiment of the present invention is described below in conjunction with fig. 1-6.
As shown in fig. 1 to fig. 6, the waveguide design 1000 for the on-chip fluorescence dispersion optical path of the flow cytometer according to the embodiment of the present invention can be understood as an on-chip fluorescence dispersion optical path structure for the flow cytometer, which can complete the fluorescence detection function of the spectral flow cytometer on a chip, and is suitable for a miniaturized and miniaturized flow cytometer system.
The waveguide design 1000 for the on-chip fluorescence dispersion optical path of the flow cytometer in the embodiment of the present invention includes a prism group 1 and a waveguide array 2, wherein the prism group 1 is used for performing dispersion separation on original fluorescence emitted by a cell in the on-chip fluorescence dispersion optical path of the flow cytometer to output fluorescence of which each wavelength has been separated; the waveguide array 2 is arranged behind the separated fluorescent light with each wavelength, the light with different wavelengths in the separated fluorescent light with each wavelength is guided into different waveguides 201 of the waveguide array 2, the light with different wavelengths is further separated in space through the bending trend of the different waveguides 201 in the waveguide array 2, higher linear dispersion rate is realized in a small-size range, and the adaptation requirement of PMT or other photoelectric detection devices is met.
In flow cytometry, the fluorescence signal emitted by the cell is detected, and the fluorescence intensity emitted by the cell is weak, so that a photomultiplier tube (PMT) or other photoelectric detection devices are used for detection. The photomultiplier tube tends to have a large spatial size, resulting in a large spatial distance between different wavelength channels, and therefore, in order to adapt to this spatial size, the dispersive optical path system must achieve a large linear dispersion ratio. The prism group 1 has a limited angular dispersion rate, and in order to achieve a sufficient linear dispersion rate and simultaneously enable the flow cytometer to meet the requirements of miniaturization and miniaturization, the waveguide array 2 is arranged behind the fluorescence with separated wavelengths emitted by the prism group 1, the light with different wavelengths in the fluorescence with separated wavelengths is guided into different waveguides 201 of the waveguide array 2, the light with different wavelengths is further separated in space through the bending trend of the different waveguides 201 in the waveguide array 2, a higher linear dispersion rate is realized in a small-size range, and the adaptation requirements of a PMT or other photoelectric detection devices are met.
The waveguide design 1000 for an on-chip fluorescence-dispersive optical path for a flow cytometer according to embodiments of the present invention has the following advantages: the introduction of the array waveguide 201 can ensure that enough dispersion rate can be still realized under small size, the fluorescence detection function of the spectral flow cytometer can be completed on a chip, the volume size of the flow cytometer can be effectively reduced, the integration of a flow cytometer system is facilitated, and the requirements of miniaturization and microminiaturization of the flow cytometer system are met; the light path stability is strong, only need general horizontally mechanical location and fixed can work, need not frequent calibration in the use.
According to some embodiments of the present invention, as shown in fig. 1 and fig. 6, the waveguide design 1000 for on-chip fluorescence dispersion optical path of flow cytometer further includes a first convex lens 3, the first convex lens 3 is disposed behind the optical path of the prism group 1 and close to the last prism in the prism group 1, and is used for converging the lights with different wavelengths in the fluorescence that has been separated by each wavelength at the position of different waveguide inlets 2011 of the waveguide array 2, so as to converge all the fluorescence that has been separated by each wavelength at the position of different waveguide inlets 2011 as much as possible.
According to some embodiments of the present invention, as shown in fig. 1 and 6, the prism assembly 1 includes a plurality of triangular prisms 101, and the plurality of triangular prisms 101 are densely arranged in an arc shape, which can reduce the space volume occupied by the prism assembly 1.
According to some embodiments of the present invention, since the waveguide design 1000 for the on-chip fluorescence dispersion optical path of the flow cytometer of the embodiments of the present invention is a planar structure, the fluorescence optical path does not have to be crossed in a plane regardless of the spatial layout and considering the size of the spatial dimension of the device, and therefore, the emission direction of the fluorescence whose wavelengths have been separated is totally deflected by 180 degrees at most with respect to the incident direction of the original fluorescence.
According to some embodiments of the present invention, as shown in fig. 1 to 6, different waveguide inlets 2011 in the waveguide array 2 are sequentially and closely arranged in a dense manner, and different waveguide outlets 2012 in the waveguide array 2 are sequentially and dispersedly arranged, so as to achieve a higher linear dispersion ratio in a small size range, and meet the adaptation requirement of the photodetector. That is, this embodiment can achieve a sufficient line dispersion ratio while allowing the flow cytometer to meet the demand for miniaturization and miniaturization.
According to some embodiments of the present invention, as shown in fig. 4 and 5, the waveguide design 1000 for the on-chip fluorescence dispersion optical path of the flow cytometer further includes a second convex lens 4, the second convex lens 4 is disposed behind different waveguide outlets 2012 in the array of waveguides 201 in a one-to-one correspondence, and converges light exiting the waveguides 201 for subsequent photodetectors to detect fluorescence, and converging light exiting the waveguides 201 with the second convex lens 4 facilitates increasing the intensity of fluorescence entering the photodetectors, thereby facilitating increasing the detection sensitivity of the micro flow cytometer.
Optionally, one second convex lens 4 may be disposed behind the exit of each single waveguide 201, or two second convex lenses 4 or lens groups may be disposed behind the exit of each single waveguide 201, so as to achieve a good detection effect. Specifically, the lens group refers to a plurality of lenses distributed along a straight line, wherein the distance between adjacent lenses and the focal length of the lenses are both subjected to certain optical design so as to meet the use requirements.
According to some embodiments of the present invention, the waveguide design 1000 for the on-chip fluorescence dispersion optical path of the flow cytometer employs a mold (as shown in fig. 7) processed on a silicon chip and a waveguide 201 is processed by one-step reverse molding of a waveguide material, the top and the bottom of the waveguide 201 are both provided with a reflective layer 6 for limiting light, and the reflective layer 6 at the bottom of the waveguide 201 is solidified inside the waveguide material, so that the processing is convenient, and the miniaturization processing is facilitated. The scheme (as shown in fig. 8) of processing the waveguide 201 with silicon dioxide or other optical dispersion materials as a structure on the wafer substrate 5 by microelectronic and MEMS processing technology can also be adopted, which is convenient for processing and is beneficial to miniaturization processing. The specific processing process comprises the following steps: firstly, a reflecting layer 6 is pasted on a wafer substrate 5, then a waveguide layer is deposited on the reflecting layer 6, then the redundant part is etched to form a waveguide 201, and finally, another reflecting layer 6 is pasted above the waveguide 201. For both embodiments, the reflective layer 6 above and below the waveguide 201 serves to limit the light exiting the optical path; the two sides of the waveguide 201 are air, and total reflection can be naturally formed by utilizing the refractive index difference between the air and the waveguide material, so that the effect of limiting light rays, which is the same as the optical fiber principle, is realized.
A specific example of a waveguide design 1000 for an on-chip fluorescence-dispersive optical path for a flow cytometer in accordance with an embodiment of the present invention is described below.
In this particular example, a waveguide design 1000 for an on-chip fluorescence dispersive optical path for a flow cytometer includes a prism assembly 1, a waveguide array 2, a first convex lens 3, and a second convex lens 4.
The prism group 1 is used for performing dispersion separation on original fluorescence emitted by cells in an on-chip fluorescence dispersion optical path of the flow cytometer to output fluorescence with separated wavelengths, and the prism group 1 comprises a plurality of triangular prisms 101, and the triangular prisms 101 are densely arranged into an arc shape, so that the size and space can be saved.
The first convex lens 3 is disposed behind the optical path of the prism group 1 and near the last prism in the prism group 1, and is used for converging light of different wavelengths in the fluorescence light whose respective wavelengths have been separated at the positions of different waveguide inlets 2011 of the waveguide array 2. The waveguide array 2 is arranged behind the separated fluorescent light with each wavelength, specifically, the first convex lens 3 converges and guides the light with different wavelengths in the separated fluorescent light with each wavelength into different waveguides 201 of the waveguide array 2 through the first convex lens 3, and the light with different wavelengths is further separated on space through the curved trend of the different waveguides 201 in the waveguide array 2, so that higher linear dispersion ratio is realized in a small-size range, and the adaptation requirement of the photoelectric detector is met. Different waveguide inlets 2011 in the waveguide array 2 are sequentially arranged closely and densely, and different waveguide outlets 2012 in the waveguide array 2 are sequentially distributed, so that higher linear dispersion rate is realized in a small-size range, and the adaptation requirement of the photoelectric detector is met.
The second convex lenses 4 are arranged behind the outlets 2012 of the different waveguides in the array of waveguides 201 in a one-to-one correspondence, and two second convex lenses 4 are arranged behind the outlet of each single waveguide 201 to converge the light emitted from the waveguides 201 so as to detect fluorescence by the subsequent PMT, thereby ensuring good detection effect.
Since the waveguide design 1000 of the on-chip fluorescence dispersion optical path for the flow cytometer of this example is a planar structure, the fluorescence optical path does not appear to cross in plane regardless of the spatial layout, and therefore, the emission direction of the fluorescence whose wavelengths have been separated is totally deflected by 180 degrees at most with respect to the incident direction of the original fluorescence.
The waveguide design 1000 for the on-chip fluorescence dispersive optical path of the flow cytometer of this example employs machining a mold on a silicon wafer and one-shot molding by back-molding.
The waveguide design 1000 for the on-chip fluorescence dispersive optical path of the flow cytometer of this example has the following advantages: the introduction of the array waveguide 201 can ensure that enough dispersion rate can be still realized under small size, the fluorescence detection function of the spectral flow cytometer can be completed on a chip, the volume size of the flow cytometer can be effectively reduced, the integration of a flow cytometer system is facilitated, and the requirements of miniaturization and microminiaturization of the flow cytometer system are met; the light path stability is strong, only need general horizontally mechanical location and fixed can work, need not frequent calibration in the use.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an illustrative embodiment," "an example," "a specific example," or "some examples" or the like are intended to mean that a particular feature, structure, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
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