Gold strip reader and detection method
Technical field
The present invention relates to gold label test strip, particularly a kind of gold strip reader and detection method.
Background technology
The solid phase labelling immuno-chromatographic assay technology of gold-marking immunity chromatographic technique to be a kind of with nanogold particle be label.Because of its high specificity, highly sensitive, fast easy, the fields such as physianthropy, farming and animal husbandry, food security and environmental monitoring are widely used in.In recent years, domestic Duo Jia 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 first obtains the measured value T of detection zone on gold label test strip and the measured value C of quality control band, and then calculate T/C, then T/C is brought in the built-in paced work curve of instrument to obtain the concentration of the checking matter dripped on detected gold label test strip.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 gold label test strip to measure, and adopts receiving light path to collect gold label test strip reflected light, uses photodiode detection gold label test strip reflected light signal.Imaging measurement mainly utilizes the imageing sensors such as CMOS, CCD gather gold label test strip image and carry out graphical analysis.
For the gold label test strip detecting instrument adopting scanning survey mode, because it measures gold label test strip in scanning motion process, Measuring Time is relatively long, and there is the repeatability that the disturbing factors such as vibration can affect measurement result in motion process.In addition, the scanning mechanism in instrument also can increase the complexity of apparatus structure and out of order probability.Imaging measurement adopts imageing sensor to carry out imaging to gold label test strip to be measured, adopts image processing method to carry out quantitative test to gold label test strip.Utilize image processing techniques to extract gold label test strip checking matter information to be measured, the stability of measurement result can be improved, and it is fast to have speed, without advantages such as complex mechanical structures; But the picture quality of gold label test strip directly can have influence on the detection sensitivity of instrument, brightness and contrast as the illumination uniformity on gold label test strip and imageing sensor can have a huge impact picture quality, and then have larger impact to the measurement result of gold label test strip, but not yet there is the report of the gold strip reader by adjusting to realize highly sensitive detection to the brightness and contrast of illumination uniformity on gold label test strip and imageing sensor at present.
Summary of the invention
For the deficiencies in the prior art, the object of the present invention is to provide a kind of gold strip reader, to realize the high-sensitivity detection to gold label test strip.
Technical solution of the present invention is as follows:
A kind of gold strip reader, by optical module, control module, input/output module and gold label test strip frame composition, it is characterized in that described optical module is by LED module, even tabula rasa, imaging lens and imageing sensor composition, multiple LED of described LED module are with the distribution in a center of symmetry of 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 up of multifunctional data acquisition card and computing machine, the light that described multiple LED send is radiated at the gold label test strip on gold label test strip frame through the illuminated field that described even tabula rasa produces even intensity, this gold label test strip is imaged on described imageing sensor by described imaging lens, the output terminal of described imageing sensor is connected with described computing machine through multifunctional data acquisition card, and described multifunctional data acquisition card is connected with described input/output module.
Utilize the method that above-mentioned gold strip reader detects gold label test strip to be measured, the method comprises the following steps:
1. illumination uniformity being corrected gold label test strip is placed on gold label test strip frame, starts gold strip reader, and the described illumination uniformity described in imageing sensor collection corrects the image of gold label test strip and stores;
2. the described brightness of imageing sensor intermediate value and the contrast of intermediate value is set; Parameter correction gold label test strip is placed on gold label test strip frame, the image input computing machine of the parameter correction gold label test strip described in described imageing sensor gathers, the value T of detection zone of the parameter correction gold label test strip corresponding to computer calculate image and the ratio of the value C of quality control band
3. change the brightness and contrast of imageing sensor, repeat step and 2. obtain N number of
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. be placed on gold label test strip frame by gold label test strip to be measured, described imageing sensor gathers the image of gold label test strip to be measured, and the image of gold label test strip to be measured is corrected the image of gold label test strip by computing machine divided by described illumination uniformity;
5. determine the ratio T/C of the value of the detection zone of gold label test strip to be measured and the value of quality control band, and calculate the concentration of the checking matter dripped on gold label test strip to be measured according to paced work opisometer.
The output module that described input/output module mainly comprises the load module for receiving user's input and exports for result.
It is blank gold label test strip that described illumination uniformity corrects gold label test strip, and the physical characteristics of blank gold label test strip is identical with the physical characteristics of gold label test strip to be detected.
Described parameter correction gold label test strip is the gold label test strip reacted, its quality control band is enriched more nanogold particle so that quality control band shows medium signal, its detection zone is enriched the nanogold particle of minute quantity so that detection zone shows fainter signal.
The present invention has following technique effect with compared with first technology:
(1) gold strip reader of the present invention is provided with illumination uniformity and corrects gold label test strip and parameter correction gold label test strip, is carried out the brightness and contrast of the imageing sensor described in accurate adjustment, to improve detection sensitivity by the image of parameter correction gold label test strip.
(2) the present invention eliminates the impact of illumination unevenness from hardware and software two aspect.In hardware design, multiple LED described in the present invention are symmetrical centered by described imaging lens, thus illumination uniformity is considered in the distribution of light source, and then, even tabula rasa is adopted to carry out homogenising to the emergent light of multiple LED, thus the homogenising again illumination intensity on gold label test strip surface.On Software for Design, the present invention corrects the image of gold label test strip by first gathering illumination uniformity, when detecting gold label test strip to be measured, the view data of gold label test strip to be measured by comparison and the view data of illumination uniformity correction gold label test strip realize, to the further optimization of illumination uniformity, improving the sensitivity of detection with this.Substantially the impact of illumination unevenness on testing result can be eliminated by the improvement of above-mentioned hardware and software two aspect.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of a kind of high sensitivity gold strip reader of the present invention.
Fig. 2 is the process flow diagram of control software design in the present invention.
Fig. 3 is the schematic diagram that illumination uniformity of the present invention corrects gold label test strip.
Fig. 4 is the schematic diagram of parameter correction gold label test strip of the present invention.
Fig. 5 is the curve that the gold label test strip 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
Below in conjunction with drawings and Examples, the present invention is further illustrated, but should not limit protection scope of the present invention with this.
Fig. 1 is the schematic diagram of gold strip reader of the present invention.As seen from the figure, gold strip reader of the present invention, by optical module 1, control module 2, input/output module 3 and gold label test strip frame 5 form, described optical module 1 is by LED module 101, even tabula rasa 102, imaging lens 103 and imageing sensor 104 form, multiple LED of described LED module 101 to be centrosymmetric distribution with 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 up of multifunctional data acquisition card 201 and computing machine 202, the light that described multiple LED send is radiated at the gold label test strip on gold label test strip frame 5 through the illuminated field that described even tabula rasa 102 produces even intensity, this gold label test strip is imaged on described imageing sensor 104 by described imaging lens 103, the output terminal of described imageing sensor 104 is connected with described computing machine 202 through multifunctional data acquisition card 201, and described multifunctional data acquisition card 201 is connected with described input/output module 3.
The effect of described multifunctional data acquisition card 201 is, the input of the input/output module 3 described in receiving on the one hand, and result is exported by described input/output module 3.Described computing machine 202 controls the image that described multifunctional data acquisition card 201 gathers described imageing sensor 104, and carries out image procossing and calculating by the control software design in computing machine.
Described control software design works as follows: 1. gather described illumination uniformity and correct the image of gold label test strip 4 and store; 2. the brightness and contrast arranging described imageing sensor 104 arranges the value of scope center for it; 3. the image of described parameter correction gold label test strip 5 is gathered, and the ratio of the value of detection zone corresponding to computed image and the value of quality control band
change the value of brightness and contrast respectively near the positive intermediate value that 4. scope be set the brightness and contrast of imageing sensor and with this value, imageing sensor arranged, now, repeating step and 3. can obtain under the arranging of current brightness and contrast
value.If be provided with the brightness and contrast that N group is different, then obtain N number of
value, can be recorded as
wherein i=1,2 ..., N; 5. N number of
in look for its maximal value and be recorded as
and with the brightness and contrast of the brightness and contrast set by now for the imageing sensor described in final, and described imageing sensor is set with this brightness and contrast; 6. gather the image of gold label test strip to be measured, the image of gold label test strip to be measured is corrected the image of gold label test strip divided by described illumination uniformity, to eliminate the impact of illumination unevenness on testing result; 7. calculate the ratio T/C of the value of the detection zone of gold label test strip to be measured and the value of quality control band, and calculate the concentration of the checking matter dripped on gold label test strip to be measured according to paced work opisometer.
Described input/output module 3 is mainly used in the instruction and the output detections result that receive user, as received the setting of user when keyboard as the present invention during load module; When testing result can show as the present invention during output module by LCD, when printer is as the printable testing result of the present invention during output module.
It is the blank gold label test strips with gold label test strip to be measured with same physical that described illumination uniformity corrects gold label test strip 4, as shown in Figure 3.When obtaining the illuminated field of even intensity on described illumination uniformity correction gold label test strip 4, the gray-scale value of each picture point in the image that described imageing sensor 104 gathers should be substantially identical, so the illumination uniformity described in utilization corrects gold label test strip 4 and carrys out correct for illumination homogeneity.
Described parameter correction gold label test strip is the gold label test strip reacted, as shown in Figure 4, the long side direction of gold label test strip is called test strips Y-direction, and gold label test strip short side direction is called test strips X-direction, its quality control band has medium signal, and its detection zone has more weak signal.
Fig. 1 is the schematic diagram of preferred embodiment, and its design parameter is as follows.The maximum absorption band wavelength of centre wavelength corresponding to the nanogold particle on gold label test strip to be measured of the LED in the LED module adopted is 525nm; The imageing sensor adopted is the cmos image sensor the most responsive to green glow, and model is 0V6630; The focal length of the imaging lens adopted is 3.7mm, and visual field is FOV43.7 × 25.8 °; The master chip of the control module adopted is the TMS320C6747 of TI company, by the EDMA data transmission module of TMS320C6747 and ping-pong double buffering technology conjunctive use, make DSP can also carry out image procossing while carrying out high-speed image sampling, drastically increase the detection speed of gold strip reader.
Fig. 2 is the process flow diagram of embodiment of the present invention control software design, can find out, in the control software design of the present embodiment, has carried out 10 times and has regulated, get wherein maximum to the brightness and contrast of imageing sensor
brightness and contrast corresponding to value is as the final settings of imageing sensor.
Fig. 5 is the curve of the present invention to the gold label test strip detection of stable reaction, the detection zone of the gold label test strip that the region that in curve, two cave in is corresponding to be measured respectively and quality control band region, region outside these two sunk areas is the region outside the functional bands of gold label test strip to be measured, is the white space of the not nano-golden particle-containing on gold label test strip to be measured.As can be seen from Figure 5, the view data change in the region outside the functional bands of gold label test strip to be measured is less, and the light intensity of the illuminated field of the gold label test strip to be measured after this explanation is calibrated is more even.
Fig. 6, Fig. 7 and Fig. 8 are to the two-dimensional measurement data that same gold label test strip to be measured detects under the crucial imaging parameters that described imageing sensor is different.The value of the brightness and contrast that Fig. 6 is corresponding is respectively: brightness settings is 0x70h, and contrast settings value is 0x68h; The value of the brightness and contrast that Fig. 7 is corresponding is respectively: brightness settings is 0xD0h, and contrast settings value is 0x68h; The value of the brightness and contrast that Fig. 8 is corresponding is respectively: brightness settings is 0x70h, and contrast settings value is 0xD0h.As seen from the figure, the sunk area that in Fig. 6, detection zone is corresponding is the most obvious, and the ratio of detection zone signal T and quality control band signal C is also maximum, so the now described brightness and contrast corresponding to imageing sensor is best settings.When adopting gold strip reader of the present invention to detect cardiac muscle troponin I (cTnI) gold label test strip, the sensitivity of detection is better than 0.25ng/ml.Meanwhile, the detection dynamic range of the present invention to cTnI gold label test strip is 0.25ng/ml ~ 64ng/ml, reaches 256 times; Being 0.134% to the coefficient of variation of high concentration gold label test strip (64ng/ml) measurement result, is 2.790% to the coefficient of variation of low concentration gold label test strip (0.25ng/ml) measurement result.
With compared with first technology, feature of the present invention is:
A kind of gold strip reader of the present invention is by optical module, control module, input/output module, and illumination uniformity corrects gold label test strip and parameter correction gold label test strip composition.When each start or when needs, illumination uniformity can be used to correct gold label test strip to improve the illumination uniformity on gold label test strip surface to be measured, and operation parameter correction gold label test strip regulates the brightness and contrast of imageing sensor to realize the high-sensitivity detection to gold label test strip to be measured.Experiment shows, the present invention can the gold label test strip to be measured to low concentration carry out highly sensitive, quantitatively detect exactly.