CN101949847B

CN101949847B - Lensless fluorescence imaging detection device and application

Info

Publication number: CN101949847B
Application number: CN2010102538326A
Authority: CN
Inventors: 金伟; 牟颖; 周超; 金钦汉
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2010-08-13
Filing date: 2010-08-13
Publication date: 2012-05-30
Anticipated expiration: 2030-08-13
Also published as: CN101949847A

Abstract

The invention provides a lensless fluorescence imaging detection device. The device consists of left and right excitation light sources, a heating plate, a microchip, an optical filter, a CCD chip and a peripheral driving circuit of the device. Compared with the common microchip detector, the lensless fluorescence imaging detection device of the invention makes full use of the characteristics of small pixel and high integration level of the CCD chip, so a corresponding relationship between a detection unit in the microchip and each pixel of the CCD chip is established by shortening the distance between the CCD chip and a reactive material, and the volume and the complexity of a detection device are greatly reduced by directly imaging without a lens. Compared with the common micro fluorescence detection device, the lensless fluorescence imaging detection device has the advantages of simple structure and operation, low requirements on a detection chip, low cost, capacity of reducing the cost and loss of the microchip, application to thermal cycling PCR reaction or isothermal nucleic acid amplification, reasonable design, simple preparation and easy operation.

Description

A kind of no lens fluorescence imaging pick-up unit and application

Technical field

The present invention relates to the fluorescence detection device of microchip, particularly a kind of device that does not need lens can carry out high throughput testing to the fluorescence signal of microchip.

Background technology

The advantage of highly integrated because of it, the low reagent loss of microchip is widely used in numerous areas such as medical diagnosis, environmental protection, drug screening.Fluoroscopic examination is the important method that is used to detect the microchip reaction result, and its ultimate principle is: excitation source shines in microchannel or the little reaction small chamber, and fluorescent material obtains energy and sends fluorescence, becomes electric signal by Computer Processing through electrooptical device.

Therefore fluorescence detection device commonly used need comprise following optical element: semi-transparent semi-reflecting lens, object lens, diaphragm etc. to the exciting light and the processing such as fluorescence filters, focusing that are stimulated.In order to improve signal quality, usually to use complicated lens group and accurate position regulating system, not only increased the volume of detection means, also improved requirement to user's operation.

Traditional imaging system all needs complicated light path system to obtain the visual field of a high enlargement ratio, and is bigger as needing the area of imaging on the fruit chip, also need introduce corresponding scanning system.The present invention adopts new formation method; Through reducing the distance between fluorescent material and the CCD chip; Its fluorescigenic scattering of institute is reduced, and phosphor dot is corresponding with one or more pixels, does not need lens also can realize the large tracts of land imaging of fluorescence reaction thing on the chip.

Summary of the invention

The purpose of this invention is to provide a kind of no lens fluorescence imaging pick-up unit; By left excitation source (1), heating plate (2), microchip (3), optical filter (4), CCD chip (5) and right excitation source (8) form, left excitation source (1) and the left and right sides parallel both sides of right excitation source (8) at microchip (3); Heating plate (2) is positioned at the below of microchip (3); Microchip (3) is made up of two-layer bonding up and down; Lower thickness is about 100 microns, and the upper strata comprises passage and detecting unit, and the upper strata of microchip (3) and optical filter (4) are close to; Optical filter (4) place CCD chip (5) under, the edge of optical filter (4) and CCD chip (5) involution.

Two sides are provided with sample interface array (6) and air valve interface array (7) over against about the microchip (3), and the position of sample interface array and air valve interface array and quantity are determined by detection microchip (3).

Left side excitation source (1) and right excitation source (8) are from two relative parallel sided incidents of detection chip (3).

Heating plate (2), its scalable temperature is 0-100 ℃, degree of regulation is 0.1 ℃, to satisfy the reaction conditions of PCR experiment.

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

The surface of microchip (3) should keep level and smooth, transparent, and its upper surface is close to optical filter (4), and to reduce the scattering that light path causes, material is selected wide, available PMMA, epoxy resin, glass, PDMS etc.

The inner structure of microchip is meticulous, and channel width and detecting unit side size range are at 10-1000 μ m, and the detecting unit quantitative range is 1-100000/mm ²

Another object of the present invention provides said device and in the PCR of thermal cycle reaction or isothermal nucleic acid amplification, uses.

Advantage of the present invention:

1. compare with microchip detecting device commonly used; No lens fluorescence imaging pick-up unit among the present invention makes full use of the characteristics that CCD chip pixel is little, integrated level is high; Through reducing the distance of CCD chip and reactive material, set up the corresponding relation of detecting unit and each pixel of CCD chip in the microchip, get final product direct imaging without lens; Significantly reduced the volume and the complexity of pick-up unit, the high flux that can be widely used in micro-fluidic chip detects in real time.

2. the no lens fluorescence imaging pick-up unit among the present invention has been eliminated because of the restriction of elements such as lens to microchip size, position, has reduced applied environment to the restriction that instrument uses, and makes microchip that more wide application space arranged.

3. the no lens fluorescence imaging pick-up unit among the present invention can detect different kinds of chips; For example polymethylmethacrylate (PMMA), polycarbonate (PC), epoxy resin, glass etc.; If adopt the malleation sample introduction, can also select dimethyl silicone polymer (PDMS) for use.

4. the resolution of the no lens fluorescence imaging pick-up unit among the present invention satisfies the detection requirement of most of microchips near the pixel size of CCD chip.

5. compare with little fluorescence detection device commonly used, structure of the present invention and simple to operate, low to the detection chip requirement, not only self is with low cost, also can reduce the cost and the loss of microchip.

The present invention reasonable in design, make simple, with low cost, easy operating.

7. this device simplicity of design need not lens combination, and detecting unit is little, signal stabilization, highly sensitive, can realize Highgrade integration, miniaturization.

Description of drawings

Fig. 1 is a device synoptic diagram of the present invention.

Fig. 2 is the synoptic diagram of microchip.

The data that Fig. 3 obtains when adopting integrated fluidic chip for microchip form images after software processes.

Embodiment

The present invention combines accompanying drawing and embodiment to be further described.

Embodiment 1

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

Referring to Fig. 2, microchip (3) is made up of two-layer up and down, and the lower thickness of microchip (3) is about 100 microns; To reduce the distance between luminescent substance and the CCD chip (5); The upper strata of microchip (3) comprises passage and detecting unit, and during detection, the upper strata of microchip (3) and optical filter (4) are close to; Optical filter (4) place between CCD chip (5) photosensitive area under; Heat by heating plate (2).Excitation source (1) parallel radiation is in the microchip side; The fluorescence signal that collects reaches computing machine through CCD chip (5) and carries out analyzing and processing.Sample interface array (6) and air valve interface array (7) are positioned at two relative sides of microchip 3, and the position and the quantity of sample interface array 6 and air valve interface array 7 are determined by detection microchip (3).

Exciting light shines from two or four sides behind fiber beam splitting simultaneously; This device does not need lens combination, and the distance between microchip (3) and the CCD chip (5) only has the distance of one deck optical filter.

Referring to Fig. 2, excitation source (1) is through the one drag two splitting optical fiber, from detecting two relative parallel sided incidents of microchip (3).

Heating plate (2) scalable temperature is 0-100 ℃, and degree of regulation is 0.1 ℃, to satisfy the reaction conditions of PCR experiment.

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

It is level and smooth, transparent that the upper and lower surfaces of microchip (3) all should keep, and optical filter (4) is close on its upper strata, to reduce the scattering that light path causes.

The imaging of data after software processes that Fig. 3 obtains when adopting integrated fluidic chip for microchip in this device (3).

Claims

1. no lens fluorescence imaging pick-up unit; Form by left excitation source (1), heating plate (2), microchip (3), optical filter (4), CCD chip (5) and right excitation source (8), left excitation source (1) and the left and right sides parallel both sides of right excitation source (8) at microchip (3), heating plate (2) is positioned at the below of microchip (3); Microchip (3) is made up of two-layer bonding up and down; Lower thickness is about 100 microns, and the upper strata comprises passage and detecting unit, and the upper strata of microchip (3) and optical filter (4) are close to; Optical filter (4) place CCD chip (5) under; The edge of optical filter (4) and CCD chip (5) involution, two sides are provided with sample interface array (6) and air valve interface array (7) over against about the microchip (3), and the position of sample interface array and air valve interface array and quantity are determined by detection microchip (3); Left side excitation source (1) and right excitation source (8) are from two relative parallel sided incidents of microchip (3); Heating plate (2), temperature regulating range are 0-100 ℃, and degree of regulation is 0.1 ℃; The thickness of optical filter (4) is 1-2mm, and the length of side is greater than the imaging region 5-10mm of CCD chip (5), edge and CCD chip (5) involution.

2. a kind of no lens fluorescence imaging pick-up unit according to claim 1 is characterized in that, it is level and smooth, transparent that the surface of microchip (3) keeps, and material is selected PMMA, epoxy resin, glass or PDMS for use.

3. a kind of no lens fluorescence imaging pick-up unit according to claim 1 is characterized in that, the channel width of microchip (3) and detecting unit side size range are at 10-1000 μ m, and the detecting unit quantitative range is 1-100000/mm ²

4. a kind of application of lens fluorescence imaging pick-up unit in the PCR of thermal cycle reaction or isothermal nucleic acid amplification of not having according to claim 1.