CN1570649A - Biochip detecting method - Google Patents
Biochip detecting method Download PDFInfo
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- CN1570649A CN1570649A CN 200410014793 CN200410014793A CN1570649A CN 1570649 A CN1570649 A CN 1570649A CN 200410014793 CN200410014793 CN 200410014793 CN 200410014793 A CN200410014793 A CN 200410014793A CN 1570649 A CN1570649 A CN 1570649A
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Abstract
This invention relates to a detecting method for biology chip, which can automatic correct biological chip parameters and grey scale, and can correct apparatus by use of automatic loop. And it is to draw biological chip active zone and to divide web grids and to correct point distance, array distance and parallel to determine the point position of biological chip, to continuously soot and synthesize the images and get average value and read biological chip and its parameters to compare them with those recorded in advance in compact disk. Before using the device, it is to automatically loop and correct chip parameters and synthesize the images and to compare those with grey scale, parameters of standard chip and accordingly to be corrected.
Description
Technical field
The present invention relates to a kind of detection system of biochip, particularly its detection method.
Background technology
At present, comprise chip, matched reagent and checkout equipment at biological chips detection system, after sample to be checked and chip and matched reagent reaction, the biological information on its chip is by the identification of biochip identifier, analyze through a cover detection method, obtain a result at last.
This detection method comprised again automatic interpretation, automatically provide assay, instrument from normal moveout correction, chip coefficient from normal moveout correction etc.
This detection method mainly is with the different information of thousands of kinds that arranged point was showed according to loaded information on the biochip, point (circle) on this chip with gray-scale value as criterion, its scope is 0～225, and the relative gray-scale value of tested point is the ratio of absolute intensity value with the index point absolute intensity value of background circle.Point on the chip is to clap by special-purpose camera, forms to have pixel, removes to catch point on the picture with corresponding empty circles, and this is manual lasso, calculate gray-scale value a little.
To differentiate gray-scale value, have qualitative and quantitative two kinds.
Qualitative be these index row of drawing by experiment in advance with us of the relative gray-scale value (A) of index to be measured row critical standard grayscale value (we claim CUTOFF value at this, B) compare, as if A＞B, the positive, otherwise feminine gender.
Quantitatively be by A according to chip, two windows of B are formed, if identifier can not automatically identify two windows (being that instrument once can only be discerned a window) simultaneously, prompting frame then occurs and " please put into chip detection window A ", the gray-scale value of index in the readout window A of elder generation, with its automatic preservation, prompting " please put into chip detection window B " then, can release the concentration value of A by the point in the window B, concrete computing method are: the curve map of window B is by the gray scale of input and concentration value is drawn out and input in advance, is known." check point concentration " (one or one group of concentration value importing when it is curve plotting figure, the longitudinal axis of curve map are that gray scale, transverse axis are concentration by prior input.), " check point " (just the index point of appointment in the window B, index point can be one also can be one group) gray-scale value can change after after a while, and then can influence result of calculation, so this point might fall less than on the curvilinear equation.The visible Fig. 1 explanation of detailed process.
YB1 is the XB1 gray-scale value that " check point " is provided with in the window B, the concentration value of representative " check point ", and these values are known.YB2 " check point " gray-scale value for reading out by machine.There have between these two values to be individual poor, illustrates that collaurum is in decay.YA is the gray-scale value of index point in the window A, and is known.YB1, XB1, YB2, YA are known, and yy, xx are desired value.Relational expression is arranged: (YB1-YB2)/XB1=(YA-X)/YA, can release the value of X, release the value of yy again by relational expression x=YA-yy, the intersection point of yy and curve is exactly xx, is the index point concentration value of being asked.
Qualitative, quantitative two kinds of discriminant approaches have set in advance in software, can directly demonstrate the result as long as click to calculate.
Instrumental correction:
Identifier owing to assembling, move and factor such as internal light source decay causes error, when practical application, we will be corrected to it unified standard, its method be (manual correction):
Machine at first carries out machine and proofreaies and correct before reading chip, proofread and correct by a standard chips. at first, read its contrast gray-scale value (ratio of index point and its background), artificial then pass through conversion formula ((x-1)/k+1, (k=(x1-1)/(x2-1) annotates the mean value that x1 represents 5 actual measurements, x2 represents the gray-scale value of standard chips corresponding point) it is transferred to the gray-scale value (we know the gray-scale value of this standard chips in advance) of standard chips, it is poor so just can to eliminate between the machine of machinery compartment.
This detection method exists obvious defects, as
Manual lasso:
Manually lasso is with manually finishing, place one's entire reliance upon people's proficiency of its accuracy, add point on the chip by a series of biochemical reactions diafiltration inhomogeneity of chip film (simultaneously) after, its point might not be the round dot of rule, moreover the color on the some face is not necessarily even, and the circle that we are used to locate is the circle of rule, we just can not entangle actual point fully like this, the position that different people also might entangle a little is different, also has its background interference, the actual grey value of reflecting point really so just, thus brought very big error for result's differentiation.
Error by hand rectifies an instrument:
Because the standard chips itself that is used to rectify an instrument also can change, add by manual lasso form and determine the error that gray-scale value brings, also want hand computation correction coefficient (very tired lock) simultaneously, will bring very big influence to the accuracy of proofreading and correct like this.
What we detected is biological information, and sample (biological products) itself will be in time, factor such as environment changes, also can produce difference between batch when producing simultaneously, this all is intrinsic existence.Above-mentioned software can not solve this difference, thereby also can result's judgement be exerted an influence.
Carry out in software for qualitative, quantitative being provided with directly, because complicated various variability of chip information, the parameter that be provided with is very many, and different chip parameter differences, so very inconvenience, and two are easy to make a mistake.
Summary of the invention
Purpose of the present invention just is to overcome above-mentioned defective, designs a kind of new detection method.
Technical scheme of the present invention:
Bio-chip test method, after sample to be checked and biochip and the matched reagent reaction, biological information on the biochip is identified instrument identification, reads, detect then, it detects step and comprises instrumental correction, manual lasso, biochip coefficient correction, and its major technique is characterised in that also and comprises:
A, automatic lasso:
The effective coverage of biochip is determined on A-1, the biochip cartridge border of delimit taking;
A-2, utilize the physical message on the biochip that grid dividing is carried out in the effective coverage, determine to have and have only in each grid a biochip point;
A-3, in each grid, search for, determine the actual boundary of biochip point;
A-3-1, the dynamic difference algorithm search in carrying out are on a large scale constructed extensive frontier point set;
A-3-2, this set is screened, determine the biochip point;
A-3-3, the biochip point position of determining is proofreaied and correct, contain dot spacing, gust spacing, collimation;
A-3-4, calculate that all the other are the biochip point position of determining;
A-3-5, the result of A-3-4 is repeated the A-3-3 step;
A-3-6, obtain automatic lasso result;
A-3-7, lasso failure automatically then transfer manual lasso to and proofread and correct.
B, image synthesize:
B-1, biochip is taken 1～10 time continuously;
B-2, above-mentioned image is synthetic obtains mean value.
C, chip coefficient correction:
C-1, read the chip coefficient of biochip;
C-2, the chip coefficient of imprinting on CD proofreaied and correct when the chip coefficient of step C-1 and biochip were dispatched from the factory, and the scope of correction in allowed limits.
D, instrument are from normal moveout correction:
D-1, to the above-mentioned A of the information via on the biochip, B, C step;
D-2, the result of D-1 step and gray scale numerical value, the chip coefficient of the standard biological chip of imprinting on CD are compared, be corrected to gray scale numerical value, the chip coefficient of standard biological chip.
Advantage of the present invention and effect are that automatic lasso can seek the actual edge of chip point automatically, have got rid of background interference, have truly reflected the actual grey of point, and the error of having avoided manual operation to cause has improved the interpretation precision, has saved the time; Image taking can continuous 1～10 pictures, and it is synthetic to carry out image, and interpretation mean value significantly reduces the error that the instrument instability is brought; Put into standard chips and gray correction CD (standard chips numerical value is carved into CD), just can carry out from normal moveout correction instrument; The chip coefficient correction has guaranteed that the chip coefficient is constant all the time, avoids having improved the interpretation accuracy rate of chip because of time, environmental change and error that manufacturing variation produced.And this method is easy to operate, quick.
Description of drawings
Fig. 1---in the prior art to the figure of chip quantitative test.
Fig. 2---the chip figure of automatic lasso among the present invention.
Fig. 3---general flow chart of the present invention.
Embodiment
Shown in Fig. 2,3, after the startup instrument begins, at first carry out the automatic correction program of instrument, automatically eject the automatic correcting window of instrument when moving for the first time, put into standard chips and gray correction CD, standard chips numerical value directly is carved in the CD in the gray correction CD, so program promptly reads on the gray correction CD corresponding numerical value automatically, carry out correct operation.
Chip coefficient correction program: read the chip coefficient of biochip, the chip coefficient of imprinting on CD proofreaied and correct when the chip coefficient that reads and chip were dispatched from the factory, and the scope of correction in allowed limits.Like this, can avoid chip its gray scale of back of dispatching from the factory to change, the chip after the variation is remained on the same level of gray scale when dispatching from the factory by correction along with the time.
Image taking: pictures taken is 1～10 time continuously, carries out image then and synthesizes, and averaged can be avoided the error that may bring because of the instrument instability like this.
Automatic lasso: delimit the biochip cartridge border of taking, determine the effective coverage of biochip; Utilize the physical message on the biochip that grid dividing is carried out in the effective coverage, determine to have and have only in each grid a biochip point; In each grid, search for, determine the actual boundary of biochip point; Dynamic difference algorithm search in carrying out is on a large scale constructed extensive frontier point set; This set is screened, determine the biochip point; The biochip point position of determining is proofreaied and correct, contain dot spacing, battle array spacing, collimation; Calculate that all the other are the biochip point position of determining; Can calculate dot spacing, battle array spacing, collimation repeatedly DR position; Obtain automatic lasso result; If lasso failure automatically then transfers manual lasso to and proofreaies and correct.
According to the chip point, setting its relative gray-scale value is H, and actual grey is H
1, the background gray scale is H
2, H=H then
2/ H
1If change, then need proofread and correct the relative gray scale H=KH=KH after the correction
2/ H
1, so H is only the gray-scale value that participates in differentiation.Thus, obtain each chip point actual count average gray, obtain the actual count average gray of each chip point background, obtain the actual gray scale relatively of each chip point, proofread and correct the actual gray scale relatively of each chip by the K value.
The automatic aligning step of chip coefficient is identical therewith.
The process of instrument in normal moveout correction comprised image taking, automatic lasso and correct operation.
Steps more of the present invention are also included within the part description of prior art, in this few description, but the reference background technology.
Claims (1)
1. bio-chip test method, after sample to be checked and biochip and the matched reagent reaction, the biological information on the biochip is identified instrument identification, reads, and detects then, it detects step and comprises instrumental correction, manual lasso, biochip coefficient correction, it is characterized in that also comprising:
A, automatic lasso:
The effective coverage of biochip is determined on A-1, the biochip cartridge border of delimit taking;
A-2, utilize the physical message on the biochip that grid dividing is carried out in the effective coverage, determine to have and have only in each grid a biochip point;
A-3, in each grid, search for, determine the actual boundary of biochip point;
A-3-1, the dynamic difference algorithm search in carrying out are on a large scale constructed extensive frontier point set;
A-3-2, this set is screened, determine the biochip point;
A-3-3, the biochip point position of determining is proofreaied and correct, contain dot spacing, gust spacing, collimation;
A-3-4, calculate that all the other are the biochip point position of determining;
A-3-5, the result of A-3-4 is repeated the A-3-3 step;
A-3-6, obtain automatic lasso result;
A-3-7, lasso failure automatically then transfer manual lasso to and proofread and correct.
B, image synthesize:
B-1, biochip is taken 1～10 time continuously;
B-2, above-mentioned image is synthetic obtains mean value.
C, chip coefficient correction:
C-1, read the chip coefficient of biochip;
C-2, the chip coefficient of imprinting on CD proofreaied and correct when the chip coefficient of step C-1 and biochip were dispatched from the factory, and the scope of correction in allowed limits.
D, instrument are from normal moveout correction:
D-1, to the above-mentioned A of the information via on the biochip, B, C step;
D-2, the result of D-1 step and gray scale numerical value, the chip coefficient of the standard biological chip of imprinting on CD are compared, be corrected to gray scale numerical value, the chip coefficient of standard biological chip.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410014793 CN1259570C (en) | 2004-04-29 | 2004-04-29 | Biochip detecting method |
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|---|---|---|---|
| CN 200410014793 CN1259570C (en) | 2004-04-29 | 2004-04-29 | Biochip detecting method |
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| Publication Number | Publication Date |
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| CN1570649A true CN1570649A (en) | 2005-01-26 |
| CN1259570C CN1259570C (en) | 2006-06-14 |
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| CN 200410014793 Expired - Fee Related CN1259570C (en) | 2004-04-29 | 2004-04-29 | Biochip detecting method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439493A (en) * | 2013-08-08 | 2013-12-11 | 南京大渊生物技术工程有限责任公司 | Aptamer percolated biochip and preparation method thereof |
| WO2022178653A1 (en) * | 2021-02-23 | 2022-09-01 | 京东方科技集团股份有限公司 | Biochip image analysis method and apparatus, and computer device and storage medium |
| CN115791776A (en) * | 2022-11-04 | 2023-03-14 | 深圳市安泰自动化设备有限公司 | Method and device for determining orientation of LED chip |
-
2004
- 2004-04-29 CN CN 200410014793 patent/CN1259570C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439493A (en) * | 2013-08-08 | 2013-12-11 | 南京大渊生物技术工程有限责任公司 | Aptamer percolated biochip and preparation method thereof |
| WO2022178653A1 (en) * | 2021-02-23 | 2022-09-01 | 京东方科技集团股份有限公司 | Biochip image analysis method and apparatus, and computer device and storage medium |
| CN115791776A (en) * | 2022-11-04 | 2023-03-14 | 深圳市安泰自动化设备有限公司 | Method and device for determining orientation of LED chip |
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| Publication number | Publication date |
|---|---|
| CN1259570C (en) | 2006-06-14 |
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