Optical module system of liquid drop high-throughput screening instrument

Abstract

The utility model provides an optical module system of a liquid drop high-throughput screening instrument. The optical module system is of an integrated structure formed by combining a fluorescence excitation part and a fluorescence imaging part. The fluorescence excitation part comprises a laser excitation part, a laser collimation part and a laser convergence part; the laser excitation part comprises a first exciter, a second exciter and a third exciter; the laser collimation part comprises three groups of lenses and cylindrical mirrors, a dichroic lens group and an adjusting mirror; the laser converging part comprises a multi-band-pass dichroic lens, a multi-band-pass wave trapping plate, a stray light eliminating sleeve and a dichroic lens. The fluorescence imaging part comprises a fluorescence receiving part and a bright field imaging part. According to the optical module system provided by the utility model, a laser spot is changed from a point light spot to a line light spot, so that the accuracy of liquid drop sorting is improved; the system also has high debugability, and the spot size and the focusing position can be flexibly adjusted through the adjusting assembly.

Description

TECHNICAL FIELD

[0001] The utility model relates to micro -fluidic technique field especially, relates to a kind of optical module system of droplet high-throughput screening instrument. BACKGROUND

[0002] Microfluidic technology is a technology for precise manipulation and analysis of microfluids in micron to nanometer channels. Because of miniaturization, integration and other characteristics, microfluidic devices are often referred to as microfluidic chips, also known as lab-on-a-chip and micro-total analytical system. One of the important features of microfluidics is the unique fluid properties in the microscale environment, such as laminar flow and droplets. Microdroplet microfluidic technology is an important branch of microfluidic technology. High-throughput screening technology of microdroplets uses two immiscible liquid phases to generate dispersed microdroplets, encapsulating single cells and biochemical reagents in single dispersed droplets. After the reaction under certain conditions, the optical signal in the droplet changes, and by analyzing the optical signal, the positive cells are sorted.

[0003] However, the current high-throughput screening method mainly has two problems: on the one hand, a single wavelength laser cannot excite multiple fluorescent markers in the droplet and recover different waveband fluorescent signals, causing missed selection or wrong selection; on the other hand, the excited fluorescence is mainly a point light spot, which cannot cover the entire droplet, causing wrong selection. Therefore, improving the optical signal analysis device in liquid screening technology and improving the sorting accuracy of droplet positive cells are urgent problems to be solved. UTILITY MODEL CONTENT

[0004] In view of the problems existing in the prior art, the utility model provides an optical module system of droplet throughput screening instrument, which is an integrated structure composed of two parts of fluorescence excitation and fluorescence imaging. The fluorescence excitation part includes laser excitation, collimation and convergence, and the laser excitation part includes first, second and third excitors. The lenses, cylindrical mirrors and dichroic lens groups of the collimation path collimate and focus the laser, so that the laser spot changes from a point light spot to a line light spot. The converged path converges the laser after collimation and shaping and folding, and reflects it into the objective lens for focusing. The fluorescence imaging part includes two parts of fluorescence reception and bright field imaging.

[0005] The utility model provides an optical module system of droplet throughput screening instrument, which includes two parts of fluorescence excitation and fluorescence imaging. The fluorescence is excited by three excitors of different wavelengths, and the laser changes from a point light spot to a line light spot after collimation and convergence and focuses on the droplet.

[0006] Further, the fluorescence excitation part includes three paths of laser excitation, collimation and convergence.

[0007] The laser exciter generally uses a semiconductor or solid laser as the excitation light source. The excitation light is collimated and shaped by the optical lens group in the collimation path, and the laser spot is changed from a point spot to a line spot after shaping. After collimation, the excitation light passes through the convergence path and converges and folds into the objective lens for focusing.

[0008] Further, the laser excitation part includes first, second and third exciters.

[0009] In some ways, the excitation light source of the exciter is a semiconductor or solid laser.

[0010] In some ways, the excitation light wavelengths used by the first, second and third lasers are 488nm, 405nm and 635nm respectively, and the three exciters are arranged side by side in the first, second and third order. The laser collimation part includes three groups of lenses and cylindrical mirrors, a dichroic lens group and an adjusting mirror. The lenses and cylindrical mirrors are arranged in the order of F30 plano-convex, cylindrical mirror, F100 plano-convex from the front of the exciter. The dichroic lens group and the adjusting mirror are arranged in the order of 488nm, 405nm and 635nm from the front of the F100 plano-convex, with one adjusting mirror, T440LPXR dichroic lens, two adjusting mirrors, T510LPXR dichroic lens and one adjusting mirror.

[0011] A set of lenses is arranged in front of each laser channel, with focal lengths of 30mm (F30 plano-convex) and 100mm (F100 plano-convex) respectively. The functions of F30 plano-convex and F100 plano-convex are to collimate the laser emitted by the laser, so that the divergence angle reaches a minimum value when the laser is incident on the objective lens. The function of the cylindrical mirror is to change the laser spot from a point spot to a line spot. After collimation and shaping, each path passes through an adjusting rack with a mirror for position adjustment. The mirror has two directions of pitch angle adjustment and can flexibly adjust the position of the spot.

[0012] In some ways, the lenses and cylindrical mirrors are fixed using stepless adjusting racks. F30 plano-convex, F100 plano-convex and cylindrical mirror are fixed using stepless adjusting racks, in order to fine-tune the focal length.

[0013] Further, the laser convergence part includes a multi-bandpass dichroic lens, a multi-bandpass notch filter, a stray light elimination sleeve and a dichroic lens.

[0014] The dichroic lens group is combined according to the laser wavelength. The three lasers of different wavelengths are arranged according to their positions so that after collimation, shaping and folding, the three lasers of different wavelengths converge on the multi-bandpass dichroic lens. The lasers are then reflected by the multi-bandpass dichroic lens and the 90° mirror at the bottom of the objective lens and enter the objective lens for focusing.

[0015] Furthermore, the fluorescence imaging section includes a fluorescence receiving section and a bright-field imaging section. The fluorescence receiving section includes an optical fiber focusing interface, and the bright-field imaging section includes an objective lens, an imaging adapter lens, a reflector, and a CCD.

[0016] The positional relationship between the laser focusing section and the fluorescence imaging section is as follows: the multi-bandpass dichroic lens is installed after aligning with the T510LPXR dichroic lens. With it as the center, the objective lens is installed in front, and the multi-bandpass notch, stray light elimination sleeve, imaging adapter lens, T710LPXR dichroic lens and reflector are installed in sequence behind it. Two adjustment mirrors and fiber optic focusing interface are installed in sequence after aligning with the T710LPXR dichroic lens. The CCD is installed after aligning with the reflector.

[0017] The recovered fluorescence passes through a multi-bandpass dichroic lens and a multi-bandpass notch filter, then enters a stray light elimination sleeve. After being reflected by a dichroic lens, it is incident on the fiber optic focusing interface via two sets of adjustment mirrors. The fiber optic focusing interface is equipped with a focusing microlens with stepless fine-tuning of the focal length, which is intended to collimate the recovered fluorescence before it is connected to the fiber optic cable.

[0018] Furthermore, the objective lens has a magnification of 10×, used to magnify the object being photographed; the imaging adapter lens has a focal length of 200mm (F200), primarily focusing the image onto the CCD; the mirror reflects the imaging light; and the CCD is used for imaging. The F200 imaging adapter lens is also fixed to a set of steplessly adjustable brackets, allowing for stepless fine-tuning of the focal length. The mirror has a unidirectional elongated aperture for fine-tuning the reflection angle, and the CCD mount has a unidirectional elongated aperture for fine-tuning the imaging focal length.

[0019] The advantages of this utility model are:

[0020] The fluorescence excitation section of the optical module can emit three different wavelengths of excitation light to excite the droplets to produce different fluorescence signals. It can collimate the laser emitted by the laser, so that the laser spot changes from a point spot to a line spot, thereby improving the accuracy of droplet sorting. Attached Figure Description

[0021] Figure 1 This is a plan view of the optical module system in Example 1.

[0022] Figure 2 This is a plan view of the laser excitation and collimation section in Example 1.

[0023] Figure 3 Structure diagram of the adjusting frame in Example 1

[0024] Figure 4 Plan view of the laser converging and fluorescence receiving part in Example 1

[0025] Figure 5 Structure diagram of the optical fiber converging interface in Example 1

[0026] Figure 6 Plan view of the bright field imaging part in Example 1

[0027] Figure 7 Structure diagram of the mirror in Example 1

[0028] Figure 8 Structure diagram of the CCD in Example 1 DETAILED DESCRIPTION

[0029] The following detailed description should be read with reference to the drawings, in which like elements in different drawings are identically numbered. The drawings, which are not necessarily to scale, depict illustrative embodiments and are not intended to limit the scope of the present application.

[0030] Example 1, the optical module system of the liquid droplet high-throughput screening instrument provided by the present application

[0031] As Figure 1 , the present application provides an optical module system 1 of a liquid droplet throughput screening instrument, which comprises a fluorescence excitation 2 and a fluorescence imaging 3. The fluorescence excitation part 2 comprises three channels of laser excitation 4, collimation 5 and convergence 6; the fluorescence imaging part 7 comprises fluorescence receiving 8 and bright field imaging 9 parts.

[0032] As Figure 2 , the laser exciter 10 generally takes LED as the excitation light source, and the excitation light passes through the optical lens group 11 in the collimation channel to be collimated and shaped, and the laser spot is changed from a point spot to a line spot after shaping. After the collimated excitation light passes through the converging part 6, it converges and folds into the objective lens 36 for focusing.

[0033] The laser excitation part 4 comprises three excitors 10 of different wavelengths, which are a first exciter 13 (488 nm), a second exciter 12 (405 nm) and a third exciter 14 (635 nm).

[0034] The laser collimation part 5 includes three sets of lenses 45 and cylindrical mirrors 46, a dichroic mirror set 15 and an adjusting mirror 16. A set of lenses 13 is arranged in front of each laser channel, with focal lengths of 30 mm (F30 plano-convex) and 100 mm (F100 plano-convex) respectively. In front of the first exciter 13, F30 plano-convex 17, cylindrical mirror 18 and F100 plano-convex 19 are arranged in sequence; in front of the second exciter 12, F30 plano-convex 20, cylindrical mirror 21 and F100 plano-convex 22 are arranged in sequence; and in front of the third exciter 14, F30 plano-convex 23, cylindrical mirror 24 and F100 plano-convex 25 are arranged in sequence. After collimation and shaping, each channel passes through the adjusting mirror 16 for position adjustment. The adjusting mirror 16 is adjusted by the adjusting frame 34 with a mirror 33, and the position of the light spot can be flexibly adjusted. One adjusting mirror 26 is arranged in front of the first laser channel, two adjusting mirrors 27 and 28 are arranged in front of the second laser channel, and two adjusting mirrors 29 and 30 are arranged in front of the third laser channel. The T440LPXR dichroic mirror 31 is arranged between the adjusting mirror 26 and the adjusting mirror 27, and the T510LPXR dichroic mirror 32 is arranged in front of the adjusting mirror 29. The dichroic mirror set is combined according to the wavelengths of the three different wavelengths of laser light, and the three different wavelengths of laser light are placed according to the positions, so that the three different wavelengths of laser light are simultaneously converged on the multi-band dichroic mirror 35 after collimation, shaping and folding, and are reflected by the multi-band dichroic mirror 35 and the 90° mirror 37 at the bottom of the objective lens 36 to enter the objective lens 36 for focusing.

[0035] As Figure 4 , the laser convergence part 6 includes a multi-band dichroic mirror 35, a multi-band notch filter 38, a stray light elimination sleeve 39 and a T710LPXR dichroic mirror 40. The fluorescent imaging part 41 includes a fluorescent receiving part 42 and a bright field imaging part 43.

[0036] The fluorescent light collected by the system passes through the multi-band dichroic mirror 35 and the multi-band notch filter 38, enters the stray light elimination sleeve 40, is reflected by the T710LPXR dichroic mirror 40, and then enters the optical fiber light collection interface 49 through the adjusting mirrors 47 and 48. The optical fiber light collection interface 49 is provided with a focusing lens 50, and the focal length can be adjusted steplessly. The purpose is to collimate the collected fluorescent light and then connect it to the optical fiber.

[0037] Further, the bright field imaging part 9 includes an objective lens 36, an imaging adapter lens 51, a mirror 52 and a CCD 53.

[0038] Wherein, the objective 36 times is 10x, for amplification of the object; imaging adapter lens 51 focal length is 200mm (F200), mainly focus on the imaging of CCD53; mirror 52 for reflecting the imaging light; CCD53 for imaging. F200 imaging adapter lens 51 is also fixed on a set of stepless adjustment of the adjusting frame 34, can stepless fine focus, mirror 52 is provided with a single direction long hole 54, for fine adjustment of the angle of reflection, CCD53 fixed seat is provided with a single direction long hole 55, for fine adjustment of the imaging focal length.

Claims

1. An optical module system for a droplet high-throughput screening instrument, characterized in that, The application relates to a fluorescence excitation and fluorescence imaging integrated structure, which comprises a laser excitation part, a laser collimation part and a laser converging part.

2. The optical module system of claim 1, wherein, The first, second and third excitation devices are arranged in parallel, and the laser collimation part is sequentially arranged in front of the excitation devices in the arrangement mode of F30 flat convex, a cylindrical mirror and F100 flat convex.

3. The optical module system of claim 2, wherein, The dichroic lens group and the adjusting mirror are sequentially arranged in front of the F100 flat convex in the direction of the first, second and third excitation devices, and the arrangement mode is one adjusting mirror, a T440LPXR dichroic lens, two adjusting mirrors, a T510LPXR dichroic lens and one adjusting mirror.

4. The optical module system of claim 3, wherein, The focal length of the F30 flat convex is 30mm, and the focal length of the F100 flat convex is 100mm.

5. The optical module system of claim 4, wherein, The fluorescence imaging part comprises a fluorescence receiving part and a bright field imaging part.

6. The optical module system of claim 5, wherein, The fluorescence receiving part comprises a fiber light collecting interface, and the bright field imaging part comprises an objective lens, an imaging adapter lens, a reflecting mirror and a CCD.

7. The optical module system of claim 6, wherein, The laser converging part and the fluorescence imaging part are arranged in the following mode: the multi-band dichroic lens is arranged behind the T510LPXR dichroic lens, the objective lens is arranged in front of the multi-band dichroic lens, and the multi-band notch filter, the stray light elimination sleeve, the imaging adapter lens, the T710LPXR dichroic lens and the reflecting mirror are sequentially arranged behind the objective lens; the two adjusting mirrors and the fiber light collecting interface are sequentially arranged behind the T710LPXR dichroic lens; and the CCD is arranged behind the reflecting mirror.

8. The optical module system of claim 7, wherein, The objective lens has a magnification of 10x, and the imaging adapter lens has a focal length of 200mm.

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