CN101949847B - Lensless fluorescence imaging detection device and application - Google Patents

Abstract

The invention provides a lensless fluorescent imaging detection device, which is composed of left and right excitation light sources, a heating plate, a microchip, an optical filter, a CCD chip and its peripheral driving circuit. Compared with commonly used microchip detectors, the lensless fluorescence imaging detection device in the present invention makes full use of the characteristics of small pixels and high integration of CCD chips, and establishes a detection unit in the microchip by reducing the distance between the CCD chip and the reaction substance. The corresponding relationship with each pixel of the CCD chip can be directly imaged without a lens, which greatly reduces the volume and complexity of the detection device. Compared with commonly used micro-fluorescence detection devices, the present invention has simple structure and operation, low requirements for detection chips, not only low cost itself, but also reduces the cost and loss of microchips, and can be used in thermal cycle PCR reactions or isothermal nucleic acid amplification application. The invention has reasonable design, simple manufacture, low cost and easy operation.

Description

A lensless fluorescence imaging detection device and its application

technical field

The invention relates to a fluorescence detection device of a microchip, in particular to a device capable of high-throughput detection of the fluorescence signal of a microchip without a lens.

Background technique

Due to the advantages of high integration and low reagent loss, microchips are widely used in many fields such as medical diagnosis, environmental protection, and drug screening. Fluorescence detection is an important method for detecting the reaction results of microchips. The basic principle is: the excitation light source is irradiated into the micropipe or microreaction chamber, the fluorescent substance obtains energy and emits fluorescence, which is converted into an electrical signal by a photoelectric conversion device and processed by a computer.

Commonly used fluorescence detection devices need to filter and focus the excitation light and the excited fluorescence, so they include the following optical components: half mirror, objective lens, aperture, etc. In order to improve the signal quality, complex lens groups and precise position adjustment systems are often used, which not only increases the size of the detection device, but also increases the requirements for user operations.

Traditional imaging systems require a complex optical path system to obtain a high magnification field of view. If the imaging area on the chip is large, a corresponding scanning system needs to be introduced. The invention adopts a new imaging method, by reducing the distance between the fluorescent substance and the CCD chip, the scattering of the fluorescent light emitted by it is reduced, the fluorescent point corresponds to one or more pixels, and the fluorescence on the chip can be realized without a lens Large area imaging of reactants.

Contents of the invention

The object of the present invention is to provide a lensless fluorescent imaging detection device, which consists of a left excitation light source (1), a heating plate (2), a microchip (3), an optical filter (4), a CCD chip (5) and a right excitation The light source (8) consists of a left excitation light source (1) and a right excitation light source (8) on the left and right parallel sides of the microchip (3), the heating plate (2) is located under the microchip (3), and the microchip (3) ) is composed of upper and lower layers of bonding, the thickness of the lower layer is about 100 microns, the upper layer contains channels and detection units, the upper layer of the microchip (3) is closely attached to the optical filter (4); the optical filter (4) is placed on the CCD chip Right below (5), the edge of the optical filter (4) is sealed with the CCD chip (5).

A sample interface array (6) and an air valve interface array (7) are arranged on the left and right sides facing the microchip (3), and the positions and numbers of the sample interface array and the air valve interface array are determined by the detected microchip (3).

The left excitation light source (1) and the right excitation light source (8) are incident in parallel from two opposite sides of the detection chip (3).

The heating plate (2) can adjust the temperature from 0 to 100°C with an adjustment accuracy of 0.1°C to meet the reaction conditions of the PCR experiment.

The thickness of the optical filter (4) is 1-2 mm, the side length should be 5-10 mm longer than the imaging area of the CCD chip (5), and the edge is sealed with the CCD chip (5).

The surface of the microchip (3) should be kept smooth and transparent, and its upper surface is close to the optical filter (4) to reduce the scattering caused by the optical path. The material selection is wide, and PMMA, epoxy resin, glass, PDMS, etc. can be used.

The internal structure of the microchip is fine, the channel width and the detection unit side length range from 10-1000 μm, and the detection unit quantity ranges from 1-100000/mm ² .

Another object of the present invention is to provide the device for use in thermocycling PCR reactions or isothermal nucleic acid amplification.

Advantages of the present invention:

1. Compared with conventional microchip detectors, the non-lens fluorescence imaging detection device in the present invention makes full use of the characteristics that the CCD chip pixel is small and the integration is high, and establishes a microchip by reducing the distance between the CCD chip and the reaction substance. The corresponding relationship between the detection unit and each pixel of the CCD chip can be directly imaged without a lens, which greatly reduces the volume and complexity of the detection device, and can be widely used in high-throughput real-time detection of microfluidic chips.

2. The lensless fluorescent imaging detection device of the present invention eliminates the limitation of the size and position of the microchip due to elements such as lenses, reduces the limitation of the application environment on the use of the instrument, and makes the microchip have a wider application space.

3. The lensless fluorescent imaging detection device in the present invention can detect different types of chips, such as polymethyl methacrylate (PMMA), polycarbonate (PC), epoxy resin, glass, etc., if positive For pressure injection, polydimethylsiloxane (PDMS) can also be used.

4. The resolution of the lensless fluorescent imaging detection device of the present invention is close to the pixel size of a CCD chip, which meets the detection requirements of most microchips.

5. Compared with commonly used micro-fluorescence detection devices, the present invention has simple structure and operation, low requirements for detection chips, not only low cost itself, but also reduces the cost and loss of microchips.

6. The present invention has reasonable design, simple manufacture, low cost and easy operation.

7. The design of the device is simple, no lens group is required, the detection unit is small, the signal is stable, and the sensitivity is high, which can realize high integration and miniaturization.

Description of drawings

Figure 1 is a schematic diagram of the device of the present invention.

Figure 2 is a schematic diagram of a microchip.

Figure 3 is the image of the data obtained when the microchip uses an integrated flow chip after being processed by software.

Detailed ways

The present invention will be further described in conjunction with drawings and embodiments.

Example 1

Referring to Fig. 1, device of the present invention is made up of left excitation light source (1), heating plate (2), microchip (3), optical filter (4) and CCD chip (5) and right excitation light source (8).

Referring to Fig. 2, microchip (3) is made of two layers up and down, and the thickness of the lower layer of microchip (3) is about 100 microns, to reduce the distance between luminescent material and CCD chip (5), the upper layer of microchip (3) It includes a channel and a detection unit. During detection, the upper layer of the microchip (3) is in close contact with the optical filter (4); the optical filter (4) is placed directly below the photosensitive area of the CCD chip (5); the heating plate (2 )heating. The excitation light source (1) is irradiated in parallel to the side of the microchip; the collected fluorescence signal is transmitted to the computer through the CCD chip (5) for analysis and processing. The sample interface array (6) and the gas valve interface array (7) are located on two opposite sides of the microchip 3, and the positions and numbers of the sample interface array 6 and the gas valve interface array 7 are determined by the detected microchip (3).

The exciting light is irradiated simultaneously from two or four sides after splitting by the optical fiber; the device does not need a lens group, and the distance between the microchip (3) and the CCD chip (5) is only the distance of one layer of optical filter.

Referring to Fig. 2 , the excitation light source (1) is parallel incident from two opposite sides of the detection microchip (3) through one and two split optical fibers.

The temperature of the heating plate (2) can be adjusted from 0 to 100° C. with an adjustment accuracy of 0.1° C. to meet the reaction conditions of PCR experiments.

The thickness of the optical filter (4) is 1-2 mm, the side length should be 5-10 mm longer than the imaging area of the CCD chip, and the edge is sealed with the CCD chip (5).

The upper and lower surfaces of the microchip (3) should be kept smooth and transparent, and its upper layer is close to the optical filter (4), so as to reduce the scattering caused by the optical path.

Fig. 3 is the imaging of the data obtained when the microchip (3) in the device adopts the integrated flow circuit chip after being processed by software.

Claims (4)

1. A lensless fluorescent imaging detection device, consisting of a left excitation light source (1), a heating plate (2), a microchip (3), an optical filter (4), a CCD chip (5) and a right excitation light source (8) The left excitation light source (1) and the right excitation light source (8) are on the left and right parallel sides of the microchip (3), the heating plate (2) is located under the microchip (3), and the microchip (3) consists of two upper and lower Layer bonding structure, the thickness of the lower layer is about 100 microns, the upper layer contains channels and detection units, the upper layer of the microchip (3) is closely attached to the optical filter (4), and the optical filter (4) is placed in the CCD chip (5) Directly below, the edge of the optical filter (4) is sealed with the CCD chip (5), and the left and right sides facing the microchip (3) are provided with a sample interface array (6) and an air valve interface array (7). The position and quantity of the interface array and the gas valve interface array are determined by the detected microchip (3); the left excitation light source (1) and the right excitation light source (8) are incident in parallel from two opposite sides of the microchip (3); the heating plate ( 2), the temperature adjustment range is 0-100°C, and the adjustment accuracy is 0.1°C; the thickness of the optical filter (4) is 1-2mm, and the side length is 5-10mm longer than the imaging area of the CCD chip (5). The chip (5) is sealed. 2. A lensless fluorescence imaging detection device according to claim 1, wherein the surface of the microchip (3) remains smooth and transparent, and the material is PMMA, epoxy resin, glass or PDMS. 3. A lensless fluorescence imaging detection device according to claim 1, characterized in that the channel width of the microchip (3) and the side length of the detection unit are in the range of 10-1000 μm, and the number of detection units ranges from 1-100000 /mm ² . 4. The application of a lensless fluorescence imaging detection device according to claim 1 in thermal cycle PCR reaction or isothermal nucleic acid amplification.

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