CN202216907U - Biochip fluorescence micro-spectrum detection device - Google Patents

Abstract

Provided is a fluorescence micro-spectrum detection device of a biological chip. (1) is an upper cover chip, (2) is a lower carrier chip, and (11) is a micro-channel filled by biological micro-fluid to be detected. Optical excitation units (12 and 13) are arranged on the upper side and the lower side of the micro-channel (11) respectively, and optical detection units (14 and 15) are symmetrically distributed on the left and the right of the micro-channel (11). Light sent out from an excitation light source (3) is filtered at the position of an optical filter (4), and the selected excitation light is focused through an optical micro-lens (5) at the top of one of the optical excitation units, is propagated in the micro-channel (11), and irradiates the biological micro-fluid to be detected. Fluorescence of materials to be detected is excited by the excitation light and collected by an optical micro-lens (7) at the top of a unit to be detected, and the fluorescence filtered by the optical filter is received by a semiconductor photoelectric conversion device (9) and converted into electrical signals to be output. The fluorescence micro-spectrum detection device of the biological chip integrates all non-electric elements of spectrum detection, is tiny in structure, and achieves miniaturization of a chip detection device.

Description

Biochip fluorescence micro-spectrum detection device

technical field

The invention relates to a biochip fluorescence microspectrum detection device, which is mainly used for the detection of microfluid fluorescence microspectrum in a biochip, and belongs to the fields of biology, analytical chemistry and medical detection. the

technical background

Biochip is a high-tech that has developed rapidly in the field of life science in recent years. It can realize the entire bioengineering analysis process and the entire laboratory function on the basis of miniaturization. It integrates sampling, dilution, reagent addition, reaction, separation and Detection and other functions are integrated in a chip, so it is popularly called "lab-on-a-chip". Its scientific and advanced nature is concentrated in two aspects: structural miniaturization and functional integration. There are four basic points in biochip technology: chip preparation, sample preparation, biochemical reaction and signal detection. Signal detection is an important part of biochip technology, which mainly includes three parts: signal generation, signal collection and transmission, signal processing and identification. Among various biochip signal detection methods, fluorescence microspectral detection has the advantages of good repeatability, strong selectivity, high sensitivity, and non-destructive detection, and is currently one of the most widely used detection technologies. Such as micro-detection of various fluorescent biomedical microfluidic chips, micro-detection of various pathogenic biomedical microarrays, calibration and quantitative micro-detection of various colloidal gold, qualitative and quantitative micro-detection of various solid-phase chemical test papers, etc. the

At present, the excitation light source used in microspectral detection technology is a traditional packaged semiconductor light emitting device, such as a semiconductor light emitting diode (LED) or a semiconductor laser diode (LD). The fluorescence micro-spectrum detectors all directly use traditional packaged photocells, such as photomultiplier tubes (PMTs) or charge-coupled devices (CCDs). For example, in 2008, Shanghai Spectrum Instrument Co., Ltd. and the Institute of Microanalysis Devices of the Department of Chemistry of Zhejiang University jointly developed the "Laser Induced Fluorescence Detection Microfluidic Chip Biochemical Analyzer" (Analytical Chemistry Instruments and Experimental Technology, 36(1): 127～ 131, 2008.1), "Portable Fluorescence Detection System Integrated with Disposable Microfluidic Chip" by Chen Xing et al. (Nanotechnology and Precision Engineering, 7(2): 127-131, 2009.3), "Microfluidic Chip Analytical Chemistry Laboratory" (Chemical Journal of Chinese Universities, 30(3): 433～445, 2009.3), "Line Scan Quasi-Confocal Fluorescence Imaging" by Yang Bin et al. (Optical Precision Engineering, 18(5): 1028～1034 , 2010.5) etc. Obviously, the current general-purpose biochip fluorescence microspectral detection device has the following shortcomings:

1. Due to the large size of the photomultiplier tube or the charge-coupled device itself, and they are used separately, a supporting optical path device is required, resulting in the bulky size of the entire fluorescence micro-spectrum detection device, which is impossible to embed into a biochip . the

2. Since neither the light excitation unit nor the light receiving unit has a lens for focusing, and many optical fibers are used as the optical transmission device, it is inevitable that there will be loss of light energy during the fiber coupling process, which severely limits the improvement of the sensitivity of the entire detection device. the

3. Since an optical path composed of various optical devices and optical fibers is required for light transmission during excitation light transmission and reflected light collection, this affects the stability of the spectral detection device in actual use. the

4. Since the detection has not achieved the zero-distance contact measurement method, the microspectral detection is interfered by non-detection objects, such as the substances that make up the microchannel wall, resulting in measurement errors. the

The current status of biochip fluorescence microspectral detection is to detect chips the size of a fingernail in a peripheral device the size of a PC host. The scientific significance of the instrument manufacturing runs counter to the scientific advancement of the biochip concept. This situation greatly This hinders the improvement of biochip integration and becomes a bottleneck in the development of biochips. In 2007, Bambang Kuswandi et al. reviewed 106 documents in the field of biochip microfluidic optical detection devices from various countries from the 1990s to 2007 in "Optical sensing systems for micro-fluidic devices: A review" , pointed out that based on MEMS (Micro Electro Mechanical System) microfabrication technology, the study of miniaturized integrated circuits of spectroscopy detection devices is the development direction of biochip microfluidic optical detection devices. The micro-spectral detection device required by biotechnology has become one of the hotspots of current research in various countries. the

Contents of the invention

The object of the present invention is to realize chip integration by providing a biochip fluorescence micro-spectrum detection device, which integrates three parts of signal detection, signal generation, signal collection and transmission, signal processing and identification. the

The present invention is realized by adopting the following technical means:

A biochip fluorescence micro-spectrum detection device, 1 is the upper cover chip, 2 is the lower chip carrier; 11 is a microchannel filled with biological microfluids to be detected; on the upper and lower sides of the microchannel 11, there are light excitation units 12 and 13, photodetection units 14 and 15 are distributed symmetrically around the microchannel 11; the photoexcitation unit is tubular, and consists of an excitation light source 3, an excitation light filter 4, an optical microlens 5 of the excitation unit and the entire excitation unit It consists of a multi-layer film 6 surrounded by four sides; the light detection unit is tubular, and consists of an optical microlens 7 of the detection unit, a detection light filter 8, a photoelectric conversion device 9 and a multi-layer film surrounded by the entire detection unit 10 components; the light emitted from the excitation light source 3 is filtered at the filter 4, and the selected excitation light is focused through the optical microlens 5 on the top of the excitation unit, and propagates into the microchannel 11 to irradiate the biological microfluid of the object to be detected The substance to be detected is excited by the excitation light to generate fluorescence, which is collected by the optical microlens 7 at the top of the detection unit. After passing through the filter 8, the filtered fluorescence is received by the semiconductor photoelectric conversion device 9 and becomes an electrical signal output. the

The aforementioned semiconductor photoelectric conversion device 9 is a photodiode or a silicon blue light cell. the

The transmission peak wavelength of the aforementioned excitation light is 525 nm. the

Compared with the prior art, the fluorescence micro-spectrum detection device of the present invention has the following obvious advantages and beneficial effects:

The invention integrates all non-electrical elements of spectrum detection, such as excitation light source, spectroscopic device, light gathering, transmission, light uniform beam, light collection, light detection and so on. By replacing the photomultiplier tube PMT or charge-coupled device CCD that cannot be embedded in the chip, the feature size of the entire device is reduced to the order of millimeters, and can even be further reduced to the order of microns. the

Description of drawings

Fig. 1 is a radial cross-sectional view of the fluorescence micro-spectrum detection device along the front-to-back direction;

Fig. 2 is a radial cross-sectional view of the fluorescence micro-spectrum detection device along the left and right directions;

Figure 3 is a schematic diagram of the overall chip structure. the

In the figure: 1 is the cover chip, 2 is the carrier chip, 3 is the excitation light source, 4 is the excitation light filter, 5 is the optical microlens of the excitation unit, 6 is the multi-layer film of the excitation unit, 7 is the optical lens of the detection unit Micro lens, 8 is a detection optical filter, 9 is a photoelectric conversion device, 10 is a multi-layer film of a detection unit, 11 is a microchannel, 12, 13 are light excitation units, 14, 15 are light detection units. the

Detailed ways

This embodiment will be described in detail below in conjunction with accompanying drawings 1 to 3 . the

The schematic diagram of the structure of this embodiment is shown in Figures 1 and 2. Fig. 1 is a radial cross-sectional view along the front-to-back direction, which consists of two light excitation units, two light detection units on the left and right, and a central microchannel. 1 and 2 in the figure are the upper cover chip 1 and the lower chip carrier 2, which are made of organic glass (PMMA). The upper cover chip is about 2mm thick, and the lower chip carrier is about 3.5mm thick. 3, 4, 5, and 6 form a light excitation unit, wherein 3 is an excitation light source, which can use a semiconductor light-emitting diode (LED) or a semiconductor laser diode (LD), and the peak wavelength of the light emitted is 475nm; 4 is an excitation light filter, The peak wavelength of transmission is also 475nm; 5 is the optical microlens of the excitation unit, which focuses the light emitted by the excitation light source onto the measured biological microfluid; It can highly reflect the light waves inside the tube and completely block the light waves outside the tube. 7, 8, 9, and 10 form a light detection unit, wherein 7 is the optical microlens of the detection unit, which focuses the fluorescence with a wavelength of 525nm produced by the measured biological microfluid; 8 is a detection light filter, and the transmission peak wavelength is 525nm; 9 is a photoelectric conversion device, which can be a photodiode (PIN) or a silicon blue light battery, and the peak wavelength of photoelectric conversion is 525nm; 10 is a multi-layer film, which surrounds the entire detection unit to form a tube, and its function is the same as above . 11 is a square microchannel with a cross-sectional area of about 0.8mm×0.8mm, which is filled with biological microfluids to be detected. Generally, the photosensitive area of the photodetection device is larger than the light emitting area of the excitation light source, so the diameter of the photodetection unit is larger than that of the photoexcitation unit. The working process of the entire micro-spectrum detection device is that the light emitted from the excitation light source 3 is filtered at the filter 4, and the selected 475nm excitation light is focused by the optical microlens 5 on the top of the excitation unit and propagated into the microchannel 11. , to irradiate the biological microfluid of the object to be detected. The substance to be detected emits fluorescence by the excitation light, which is collected by the optical microlens 7 on the top of the detection unit. After passing through the optical filter 8, the filtered 525nm fluorescence is received by the semiconductor photoelectric conversion device 9 and becomes an electrical signal output. the

Fig. 2 is the radial sectional view of the detection device along the left and right direction, in which only the bottom chip 2, the microchannel 11, and the microlens 7 in the light excitation unit 3, 4, 5, 6 and the light detection unit can be drawn. On the transverse cross-sectional view of the detection device, only the lower chip 2, the microchannel 11, and the microlens 5 in the light detection units 7, 8, 9, 10 and the light excitation unit can be drawn. The purpose of using two sets of light excitation units and two sets of light detection units is to further improve the detection sensitivity. the

The fluorescence micro-spectrum detection device of the present invention has the following characteristics: it integrates all non-electrical elements of spectrum detection, such as excitation light source, spectroscopic device, light aggregation, transmission, light uniformity, light collection, light detection, etc. By replacing the photomultiplier tube PMT or charge-coupled device CCD that cannot be embedded in the chip, the feature size of the entire device is reduced to the order of millimeters, and can even be further reduced to the order of microns. the

In a specific embodiment, the photoexcitation unit and the photodetection unit are fabricated respectively. The fabrication process of the photoexcitation unit is that firstly the excitation light source and the excitation light filter are surrounded into a tubular shape with a multi-layer film, and then an in-situ The micro-optical lens is made by forming method, that is, the liquid droplet of UV-curable glue is released vertically from a certain height, drops onto the filter, and spreads and flows from top to bottom, and irradiates the droplet with ultraviolet laser in time, so that It cures, forming micro-optical lenses. The fabrication method of the light detection unit is similar to that of the light excitation unit. First, the photoelectric conversion device and the detection light filter are surrounded by multi-layer films to form a tube, and then the micro-optical lens is fabricated on the filter by in-situ molding. A hole is punched on the upper cover chip 1 , and the photoexcitation unit 12 is placed in it with the microlens facing down, as shown in FIG. 3 . On the lower chip 2, a square groove 11 is etched with a laser as a microchannel for the biological microfluid to be detected, and then three holes are punched at the bottom and both sides of the microchannel, and the light excitation unit 13 is placed in the bottom hole, Microlens facing upwards. Photodetection units 14 and 15 are respectively placed in the holes on both sides, the microlens of photodetection unit 14 is to the right, and the microlens of photodetection unit 15 is to the left, as shown in Figure 3. Finally, the upper cover chip 1 and the lower carrier chip 2 are bonded together to assemble a biochip fluorescence micro-spectrum detection device. At this time, the four micro-lenses are all facing the micro-channel flowing with the analyte. the

Claims (4)

1. fluorescence biosensor chip low-light spectrum detection device comprises: the cover core sheet (1) on top, the bottom carry chip (2), be filled with the microchannel (11) of biological microfluid to be detected; It is characterized in that: the upper and lower sides in the microchannel is provided with optical excitation unit (12) (13), and the left-right symmetric of (11) is distributed with optical detecting unit (14) (15) in the microchannel;

Described optical excitation unit is a tubulose, by excitation source (3), exciting light optical filter (4), excite the unit optical microlens (5) and with whole excite the unit around the multilayer film (6) that are surrounded form;

Described optical detecting unit is a tubulose, by the optical microlens (7) of detecting unit, detect light optical filter (8), electrooptical device (9) and the multilayer film (10) that are surrounded around the whole detecting unit are formed;

The light that sends from excitation source (3) is filtered at optical filter (4), and the exciting light of selecting focuses on through the optical microlens (5) that excites the top, unit, propagates into lining, microchannel (11), shines object organisms microfluid to be detected; Material to be detected goes out fluorescence by excitation, and the optical microlens (7) at top, unit to be detected is gathered, and behind optical filter (8), the fluorescence that leaches is received by semiconductor light power conversion device (9), becomes electric signal output.

2. fluorescence biosensor chip low-light spectrum detection device according to claim 1 is characterized in that: described semiconductor light power conversion device (9) is photodiode or silicon blue cell.

3. fluorescence biosensor chip low-light spectrum detection device according to claim 1 is characterized in that: the transmission peak value wavelength of described exciting light is 525nm.

4. the method for making of a kind of fluorescence biosensor chip low-light spectrum detection device according to claim 1 is characterized in that: described microchannel (11) are square trench.

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