Colloidal-gold strip reading apparatus and detection method
Technical field
The present invention relates to the gold test strip bar, particularly a kind of colloidal-gold strip reading apparatus and detection method.
Background technology
The gold-marking immunity chromatographic technique be a kind of be the solid phase labelling immuno-chromatographic assay technology of label with the nanogold particle.Because of its high specificity, highly sensitive, easy to be quick, fields such as physianthropy, farming and animal husbandry, food security and environmental monitoring have been widely used in.In recent years, domestic how tame unit had carried out the research of quantitative gold-marking immunity chromatographic technique in succession, and developed relevant golden scalar quantity detecting instrument.For golden scalar quantity detecting instrument, its quantitative testing process normally obtains detecting on the gold test strip bar measured value T of band and the measured value C of quality control band earlier, and then calculate T/C, again T/C is brought in the built-in paced work curve of instrument to obtain dripping the concentration of the checking matter on detected gold test strip bar.The detection mode of gold scalar quantity detecting instrument mainly contains two kinds: scanning survey and imaging measurement.Scanning survey adopts mechanical driving device to drive the gold test strip bar and measures, and adopts receiving light path to collect gold test strip bar reflected light, uses photodiode to survey gold test strip bar reflected light signal.Imaging measurement mainly utilizes imageing sensors such as CMOS, CCD to gather gold test strip bar image and carries out graphical analysis.
For the gold test strip bar detecting instrument that adopts the scanning survey mode, because it is measured the gold test strip bar in the scanning motion process, Measuring Time is longer relatively, and exists disturbing factor such as vibration can influence the repeatability of measurement result in the motion process.In addition, the scanning mechanism in the instrument also can increase complexity and the out of order probability of apparatus structure.Imaging measurement adopts imageing sensor that gold test strip bar to be measured is carried out imaging, adopts image processing method that the gold test strip bar is carried out quantitative test.Utilize image processing techniques to extract gold test strip bar checking matter information to be measured, can improve the stability of measurement result, and it is fast to have speed, advantages such as no complex mechanical structure; But the picture quality of gold test strip bar can directly have influence on the detection sensitivity of instrument, brightness and contrast as the illumination uniformity on the gold test strip bar and imageing sensor can produce very big influence to picture quality, and then the measurement result of gold test strip bar had bigger influence, but the report of the colloidal-gold strip reading apparatus of highly sensitive detection does not appear adjusting to realize by the brightness and contrast to illumination homogeneity and imageing sensor on the gold test strip bar at present as yet.
Summary of the invention
At the deficiencies in the prior art, the object of the present invention is to provide a kind of colloidal-gold strip reading apparatus, to realize the high-sensitivity detection to the gold test strip bar.
Technical solution of the present invention is as follows:
A kind of colloidal-gold strip reading apparatus, by optical module, control module, input/output module and gold test strip bar frame are formed, it is characterized in that described optical module is by the LED module, even tabula rasa, imaging lens and imageing sensor are formed, a plurality of LED of described LED module become the center to be symmetrically distributed with described imaging lens, the center of described even tabula rasa has circular hole and stretches out for described imaging lens, described control module is made of multifunctional data acquisition card and computing machine, and the light that described a plurality of LED send produces the uniform illuminated field of intensity through described even tabula rasa and is radiated at gold test strip bar on the gold test strip bar frame; This gold test strip bar is imaged on the described imageing sensor by described imaging lens; The output terminal of described imageing sensor links to each other with described computing machine through multifunctional data acquisition card, and described multifunctional data acquisition card links to each other with described input/output module.
The method of utilizing above-mentioned colloidal-gold strip reading apparatus that gold test strip bar to be measured is detected, this method comprises the following steps:
1. illumination uniformity is proofreaied and correct the gold test strip bar and placed on the gold test strip bar frame, start the colloidal-gold strip reading apparatus, described imageing sensor is gathered image and the storage that described illumination uniformity is proofreaied and correct the gold test strip bar;
2. set the brightness of described imageing sensor intermediate value and the contrast of intermediate value; Parameter correction gold test strip bar is placed on the gold test strip bar frame, described imageing sensor is gathered the image input computing machine of described parameter correction gold test strip bar, the ratio of the value T of the detection band of the corresponding parameter correction gold test strip of computing machine computed image bar and the value C of quality control band
3. change the brightness and contrast of imageing sensor, 2. repeating step obtains N
Value is recorded as respectively
Wherein i=1,2 ..., N; Relatively find out maximal value and be recorded as
The brightness and contrast of max arranges described imageing sensor;
4. gold test strip bar to be measured is placed on the gold test strip bar frame, described imageing sensor is gathered the image of gold test strip bar to be measured, and the image of gold test strip bar proofreaied and correct the image of gold test strip bar to be measured by computing machine divided by described illumination uniformity;
5. determine the ratio T/C of the value of the value of detection band of gold test strip bar to be measured and quality control band, and calculate the concentration of the checking matter of dropping on gold test strip bar to be measured according to the paced work opisometer.
Described input/output module mainly comprises for the load module that receives user's input and is used for the output module that the result exports.
It is blank gold test strip bar that described illumination uniformity is proofreaied and correct the gold test strip bar, and the physical characteristics of the physical characteristics of blank gold test strip bar and gold test strip bar to be detected is identical.
Described parameter correction gold test strip bar is to have reacted the gold test strip bar of finishing, on its quality control band enrichment more nanogold particle so that quality control band shows medium signal, its detection with go up enrichment the nanogold particle of minute quantity show fainter signal so that detect band.
The present invention compares with technology formerly has following technique effect:
(1) colloidal-gold strip reading apparatus of the present invention is provided with illumination uniformity and proofreaies and correct gold test strip bar and parameter correction gold test strip bar, and the brightness and contrast that the image by parameter correction gold test strip bar comes the described imageing sensor of accurate adjustment is to improve detection sensitivity.
(2) the present invention eliminates the influence of illumination unevenness from hardware and software two aspects.On hardware design, a plurality of LED described in the present invention are centered by described imaging lens and be symmetrically distributed, thereby illumination uniformity has been considered in the distribution at light source, and then, adopt even tabula rasa that the emergent light of a plurality of LED is carried out homogenising, thus homogenising again the illumination light intensity on gold test strip bar surface.In the software design, the present invention will at first gather the image that illumination uniformity is proofreaied and correct the gold test strip bar, when gold test strip bar to be measured is detected, the view data that view data by comparing gold test strip bar to be measured and illumination uniformity are proofreaied and correct the gold test strip bar realizes the inhomogeneity further optimization of throwing light on is improved the sensitivity of detection with this.Substantially can eliminate illumination unevenness to the influence of testing result by the improvement of above-mentioned hardware and software two aspects.
Description of drawings
Fig. 1 is the synoptic diagram of a kind of high sensitivity colloidal-gold strip of the present invention reading apparatus.
Fig. 2 is the process flow diagram of control software among the present invention.
Fig. 3 proofreaies and correct the synoptic diagram of gold test strip bar for illumination uniformity of the present invention.
Fig. 4 is the synoptic diagram of parameter correction gold test strip bar of the present invention.
Fig. 5 is the curve that the gold test strip bar of stable reaction detects.
Fig. 6 adopts the present invention to the 2-D data of gold designation strip detecting to be measured under a certain group of brightness and contrast.
Fig. 7 adopts the present invention to the 2-D data of gold designation strip detecting to be measured under a certain group of brightness and contrast.
Fig. 8 adopts the present invention to the 2-D data of gold designation strip detecting to be measured under a certain group of brightness and contrast.
Embodiment
The present invention is further illustrated below in conjunction with drawings and Examples, but should not limit protection scope of the present invention with this.
Fig. 1 is the synoptic diagram of colloidal-gold strip reading apparatus of the present invention.As seen from the figure, colloidal-gold strip reading apparatus of the present invention, by optical module 1, control module 2, input/output module 3 and gold test strip bar frame 5 are formed, described optical module 1 is by LED module 101, even tabula rasa 102, imaging lens 103 and imageing sensor 104 are formed, a plurality of LED of described LED module 101 are with the distribution that is centrosymmetric of described imaging lens 103, the center of described even tabula rasa 102 has circular hole and stretches out for described imaging lens 103, described control module 2 is made of multifunctional data acquisition card 201 and computing machine 202, and the light that described a plurality of LED send produces the uniform illuminated field of intensity through described even tabula rasa 102 and is radiated at gold test strip bar on the gold test strip bar frame 5; This gold test strip bar is imaged on the described imageing sensor 104 by described imaging lens 103; The output terminal of described imageing sensor 104 links to each other with described computing machine 202 through multifunctional data acquisition card 201, and described multifunctional data acquisition card 201 links to each other with described input/output module 3.
The effect of described multifunctional data acquisition card 201 is, receives the input of described input/output module 3 on the one hand, and the result is exported by described input/output module 3.The described multifunctional data acquisition card 201 of described computing machine 202 controls is gathered the image of described imageing sensor 104, and carries out image by the control software in the computing machine and handle and calculate.
Described control software carries out following work: 1. gather image and storage that described illumination uniformity is proofreaied and correct gold test strip bar 4; The brightness and contrast that 2. described imageing sensor 104 is set arranges the value of scope center for it; 3. gather the image of described parameter correction gold test strip bar 5, and the ratio of the value of the value of the corresponding detection band of computed image and quality control band
4. the brightness and contrast of imageing sensor arrange scope positive intermediate value near change brightness and contrast's value respectively and be worth with this imageing sensor is arranged, at this moment, 3. repeating step can obtain under the arranging of current brightness and contrast
Value.If be provided with the different brightness and contrast of N group, then obtained N
Value can be recorded as
Wherein i=1,2 ..., N; 5. at N
In look for its maximal value and be recorded as
And be the brightness and contrast of final described imageing sensor with the set brightness and contrast of this moment, and with this brightness and contrast described imageing sensor is set; 6. gather the image of gold test strip bar to be measured, the image of gold test strip bar to be measured is proofreaied and correct the image of gold test strip bar divided by described illumination uniformity, to eliminate illumination unevenness to the influence of testing result; 7. calculate the ratio T/C of the value of the value of detection band of gold test strip bar to be measured and quality control band, and calculate the concentration of the checking matter of dropping on gold test strip bar to be measured according to the paced work opisometer.
Described input/output module 3 is mainly used in receiving user's instruction and output testing result, as when keyboard during as load module the present invention can receive user's setting; When LCD during as output module the present invention testing result can be shown, when printer printable testing result of the present invention during as output module.
It is the blank gold test strip bars that have same physical with gold test strip bar to be measured that described illumination uniformity is proofreaied and correct gold test strip bar 4, as shown in Figure 3.When described illumination uniformity correction gold test strip bar 4 has obtained the uniform illuminated field of intensity, the gray-scale value of each picture point in the image that described imageing sensor 104 is gathered should be basic identical, so, utilize described illumination uniformity to proofread and correct gold test strip bar 4 and come the correct for illumination homogeneity.
Described parameter correction gold test strip bar is to have reacted the gold test strip bar of finishing, as shown in Figure 4, the long side direction of gold test strip bar is called the test strips Y-direction, and gold test strip bar short side direction is called the test strips directions X, its quality control band has medium signal, and it detects band and has more weak signal.
Fig. 1 is the synoptic diagram of most preferred embodiment of the present invention, and its concrete parameter is as follows.The centre wavelength of LED in the LED module that adopts is the corresponding maximum absorption band wavelength of nanogold particle on the gold test strip bar to be measured, is 525nm; The imageing sensor that adopts is the cmos image sensor the most responsive to green glow, and model is 0V6630; The focal length of the imaging lens that adopts is 3.7mm, and the visual field is FOV43.7 * 25.8 °; The master chip of the control module that adopts is the TMS320C6747 of TI company, EDMA data transmission module and the ping-pong double buffering technology of TMS320C6747 are united utilization, make DSP when carrying out high-speed image sampling, can also carry out image and handle, greatly improved the detection speed of colloidal-gold strip reading apparatus.
Fig. 2 is the process flow diagram of embodiment of the invention control software, can find out, in the control software of present embodiment, the brightness and contrast of imageing sensor has been carried out 10 times regulated, and gets wherein maximum
Be worth corresponding brightness and contrast as the final value of setting of imageing sensor.
Fig. 5 curve that to be the present invention detect the gold test strip bar of stable reaction, detection band and the quality control band zone of gold test strip bar corresponding to be measured distinguished in the zone of two depressions in the curve, zone outside these two sunk areas is the zone outside the function band of gold test strip bar to be measured, is the white space of the not nano-golden particle-containing on the gold test strip bar to be measured.As can be seen from Figure 5, the view data in the zone outside the function band of gold test strip bar to be measured changes less, and the light intensity of the illuminated field of the gold test strip bar to be measured after this explanation is calibrated relatively evenly.
Fig. 6, Fig. 7 are the two-dimensional measurement data that under the different crucial imaging parameters of described imageing sensor same gold test strip bar to be measured detected with Fig. 8.The brightness and contrast's of Fig. 6 correspondence value is respectively: brightness settings is 0x70h, and the contrast settings value is 0x68h; The brightness and contrast's of Fig. 7 correspondence value is respectively: brightness settings is 0xD0h, and the contrast settings value is 0x68h; The brightness and contrast's of Fig. 8 correspondence value is respectively: brightness settings is 0x70h, and the contrast settings value is 0xD0h.As seen from the figure, it is the most obvious to detect the corresponding sunk area of band among Fig. 6, and the ratio of detection band signal T and quality control band signal C is also maximum, so the corresponding brightness and contrast of described imageing sensor this moment is the best value of setting.When adopting colloidal-gold strip reading apparatus of the present invention that cardiac muscle troponin I (cTnI) gold test strip bar is detected, the sensitivity of detection is better than 0.25ng/ml.Simultaneously, the present invention is 0.25ng/ml~64ng/ml to the detection dynamic range of cTnI gold test strip bar, has reached 256 times; The coefficient of variation to high concentration gold test strip bar (64ng/ml) measurement result is 0.134%, and the coefficient of variation of low-concentration gold being marked test strips (0.25ng/ml) measurement result is 2.790%.
Compare with technology formerly, characteristics of the present invention are:
A kind of colloidal-gold strip reading apparatus of the present invention is by optical module, control module, input/output module, and illumination uniformity is proofreaied and correct the gold test strip bar and parameter correction gold test strip bar is formed.When each start or in needs, can use illumination uniformity to proofread and correct the gold test strip bar and improve the illumination uniformity on gold test strip bar surface to be measured, operation parameter is proofreaied and correct the gold test strip bar and is regulated the brightness and contrast of imageing sensor and realize high-sensitivity detection to gold test strip bar to be measured.Experiment shows that the present invention can carry out highly sensitive, quantitatively detection exactly to low concentration gold test strip bar to be measured.