DZ215单片机控制半自动酶标仪
DZ215单片机控制半自动酶标仪,dz215,单片机,控制,节制,半自动,酶标仪
单片机控制半自动酶标仪,指导教师:李 岩学 生:郭一男 班 级:040301班,题目的来源,近二十年来,由于生物学,近代物理学,化学,电子技术,生物材料学,机械学,计算机等多种科学的飞速发展,并愈来愈深入得对生物学和临床医学领域的广泛渗透,促进了医学检验理论急实验室仪器和技术的发展.自动化的检测仪器的涌现,大大提高了医学检验的测量精度,速度,准确度,从而使繁琐的操作得以简化,复杂的成分及微含量的测量得以实现,酶标分析仪就是在这种形式下出现的临床检测仪器,酶标仪的检测原理,酶是生物体内细胞产生的具有高效特异催化作用的蛋白质,有趣的是,一种酶只对某一种化学反应起催化作用,而对其他化学反应不起作用。这就是酶的标记原理。酶联免疫吸附测试正是利用了抗体与抗原反应的特异性、酶的标记原理和高效催化作用,所以具有高度的特异性和灵敏性。,主要技术指标,检测范围: 0-3A(405nm);精度: 读取范围 0-4.0A;准确性 1%;液晶显示,声光报警;打印数据,完成与上位机通信。,酶标仪硬件系统,酶标仪系统主要包括一下几个部分:滤波片机械操作系统数据的采集与传输单片机控制系统工作过程如下: 由光电检测器检测信号,经过放大,A/D转换,送入主机进行数据处理,判断分析,另外由单片机控制电机来实现微面板的运动,系统的总体结构,系统框图:,电机控制,控制系统的选择,控制芯片选择单片机89C51: 89C51是一种带4K字节闪烁可编程可擦除只读存储器的单片机。单片机的可擦除只读存储器可以反复擦除100次。该器件采用ATMEL高密度非易失存储器制造技术制造,与工业标准的MCS-51指令集和输出管脚相兼容。由于将多功能 8位CPU和闪烁存储器组合在单个芯片中,ATMEL的89C51是一种高效微控制器,89C51单片机为很多嵌入式控制系统提供了一种灵活性高且价廉的方案。,主要特征: 与MCS-51 兼容,4K字节可编程闪烁存储器,寿命100写/擦循环,数据保留时间:10年,全静态工作:0Hz-24Hz,三级程序存储器锁定 128*8位,内部RAM,32可编程I/O线,两个16位定时器/计数器,5个中断源,可编程串行通道,低功耗的闲置和掉电模式,片内振荡器和时钟电路。,电机控制系统,在控制系统中,选用TA8435H步进电机驱动芯片的执行机构,通过步进电机的运动来完成微面板的直线往复运动。芯片特点: 工作电压范围宽(10V40V) 输出电流可达平均为1.5A,峰值可以达到2.5A; 具有整步、半步、细分、细分运行方式可供选择; 采用脉宽调制式斩波驱动方式; 具有正反转控制功能; 带有复位和使能引脚; 可选择使用单时钟输入或双时钟输入;,电路用一片TA8435H来驱动电机,输入信号有使能控制、正反转控制和时钟输入,通过光耦可将驱动器与输入级进行电隔离,以起到逻辑电平隔离和保护作用;该电路工作在1/8细分模式(M1、M1接高电平),可减小低速时的振动. 另外采用外部定时计数器82C53来给TA8435H提供步进脉冲。 82C53的工作方式是一种方波速率发生器。在这种方式下,当CPU设置控制好后,输出将为高电平,在写完计数值后就自动开始计数,输出保持高电平;而当计到一半计数值时,输出变低直到计数到0,此后输出又变高以重新开始计数。,放大电路系统设计,系统设计要求的检测范围是0405nm,检测到的电流应在0100nA,通过一个5K电阻得到的电压值在00.0005V,在经过两级放大之后,由于一级放大的倍数为100倍,那两级放大之后,使其电压值可以达到05V,刚好符合A/D转换芯片AD976A输入电压的范围,符合系统的设计要求。,A/D转换电路设计,在A/D转换电路中,选用由AD公司生产的AD976A, AD976A是AD公司推出的l6位高速,高精度A/D转换器,最大通过率为100kSPS,输入信号范围为-l0一+I0V,带宽为1.5MHz,它可以与8位,16位单片机和DSP方便接口,使用十分方便灵活。它的主要特征如下:(1)AD976A是具有100kSPs200kSPs高通过率的并行输出ADC;(2)需+5V单电源供电,且具有100mW的低功耗:(3)片上集成有ADC、参考电源和时钟;(4)采用开关电容电荷重分布结构,其内部的自动校正逻辑可以校正内部的非线性,从而使其性能总体上得到了优化。,A/D转换电路硬件连接图:,显示与键盘系统设计,显示系统: 酶标仪具有显示功能,本设计中,主要显示的是各路采集的数据,同时显示时间日期等功能,所以用一般的LED显示可能很难实现,所以我采用LCD液晶显示,并采用PCF2111CT字符液晶驱动模块。,PCF2111CT模块的主要特点有:(1)工作电压要求范围大,DC2.25V6.0V;低功耗,最大功耗100mW;(2)内置振荡电路,无需外接振荡器,只要在振荡器输入端与电源跨接一个电容,与地跨接一个电阻即可,使用设计方便;(3)数据串行连续输入,与外围电路特别是单片机连接时,只需3根线,可以节约单片机的I/O口(4)具有加电自动复位功能;(5)具有液晶底板背光驱动功能。,液晶显示电路连接图: 39脚、40脚分别接单片机,37、38脚接发光二极管,用于液晶的背光显示。单片机的P1.0、P1.1、P1.2三脚分别充当同步写入时钟CLB、数据输入DATA、输入控制DLEN的功能。初始化单片机时置P1.2为低电平,写入数据时,置P1.2高电平。,键盘系统: 系统的键盘输入主要有0到8路数据采集的控制,复位等功能,需要的键盘数量较多,我采用82矩阵键盘,控制芯片则采用HD7279A。 HD7279A是一种管理键盘和LED显示器的专用智能控制芯片,同时能对多达88的键盘矩阵的按键情况进行监视,具有自动消除键抖动并识别按键代码的功能,从而可以提高CPU工作的效率. HD7279A和微处理器之间采用串行接口,其接口电路和外围电路简单,占用口线少,加之它具有较高的性能价格比,因此,在微型控制器、智能仪表、控制面板和家用电器等领域中日益获得广泛的应用。,软件设计,在主程序中,首先对单片机80C51等功能芯片进行初始化,设定各项初值。然后检测按键,进行按键处理。键盘处理结束后,通过电机运动,实现数据采集。,因为需要采集8路数据,所以用ch表示所采集的路段,其采集子程序流程图如图,谢谢各位老师指导, Enzyme sign meter synopsisNamely the enzyme unites the immunity instrumentation (ELISA Reader) is the enzyme unites the immunity adsorption test the special-purpose instrument. But divides into semiautomatic and the completely automatic 2 broad headings simply, but its principle of work basically is consistent, its core is a colorimeter, namely color method analyzes the antigen or the immune body content with the . ELISA determines the general request test fluid the final volume below 250 l, common photoelectric colorimeter is unable with the to complete the test, therefore has the special request to the enzyme sign meters photoelectric colorimeter.First introduced the enzyme sign meters principle of work and structure:The enzyme sign meter in fact is special-purpose photoelectric colorimeter or the spectrophotometer in disguised form, its key job principle and primary structure and photoelectric colorimeter basic same. the graphical representation is one kind of single channel automatic specimen handling enzyme sign meter work schematic diagram. The photo source lamp sends out the light wave turns bunch of monochromatic lights after the light filter or monochromator , enters plastic pore extremely the testing specimen. This monochromatic light part by the specimen absorption, another part is passed the specimen to shine to the electro-optical detector on, the electro-optical detector this testing specimen is different, but the strong and the weak different light signal transforms corresponding the electrical signal. The electrical signal after the pre-amplification, the logarithm enlarges, after a/d conversion equisignal processing, sends in the microprocessor to carry on data processing and the computation, finally demonstrates the result by the monitor and the printerThe microprocessor also moves the micro orifice through the control circuit ncontrol machinery driving mechanism X direction and the Y directions movement, thus solid is presently automatic the specimen handling examination process. But another some enzyme sign meter is uses moves the micro orifice to carry on the examination manually, therefore has omitted X, Y the direction machinery driving mechanism and the control circuit, thus causes the instrument to be more exquisite, the structure is also simpler. the micro orifice is one kind uses in after Bao Lizhuan laying aside the testing sample beforehand the transite plate, on the board has multi-platoon size even consistent the eyelet, in the hole the embedding corresponding antigen or the immune body, in the micro orifice each eyelet may Sheng Fangling select several milliliters solutions. Its common the specification has 40 orifices, 55 orifices, 96 orifices and so on many kinds, the different instrument selects the different specification the orifice, to its may carry on a examination or a row of row of place examination.The enzyme sign meter uses the monochromatic light already may obtain through the coherent light filter, also the available spectrophotometer same monochromator comes . When uses the light filter make the filter installment with the ordinary colorimeter is the same, the light filter then place the micro orifice front, may also place the micro orifice behind, its effect is the same. The next chart is the present commonly used enzyme sign meter path of rays diagram. The photo source lamp sends out light after condensing lens, after light fence, arrives at the reflector, makes the 90o reflection after the reflector is vertical through compares the color solution, then passes through the light filter to deliver again the electric eye.It and the ordinary photoelectric colorimeter has the following several differences:(1) Splendid attire testing no longer uses compared to the color fluids vessel compares the color dish, but uses the plastic micro orifice. Micro orifice commonly used transparent the polyethylene material makes, has the strong adsorption to the antigen immune body, therefore takes the solid phase carrier with it. (2) Because the abundant samples plastic micro orifice is the multi-platoons porous, the optical fiber can only vertical pass through, therefore the enzyme sign meters light comes is vertical through the testing solution and the micro orifice, but light beam already from top to bottom, may also be passes through from bottom to top compares the color fluid. not only the (3) Enzyme sign meter usually uses A, sometimes also uses light density OD to express the extinction. the enzyme sign meter may divide into the single channel and the multichannel 2 types, the single channel also has automatic and the manual 2 kind of division. Automatic instrument has the X, Y direction machinery driving mechanism, may one each one send in the micro orifice L eyelet under the light beam to test in turn, manual, then depending on moves the micro orifice to carry on the survey manually.Has developed the multichannel enzyme sign in the single channel enzyme sign meters foundation only, this kind of enzyme sign only generally is the automatic molding. It does not have many light beams and many electro-optical detectors, if 12 channels instruments are equipped with 12 light beams or 12 photo sources, 12 detectors and 12 amplifiers, under X directions machinery drives function, sample 12 are one row are examined. The multichannel enzyme sign meters examination speed is quick, but its structure complex price is also highThe enzyme unites the immunity adsorption test method:The enzyme sign I the immunity adsorption test method abbreviation enzyme sign law, is in the labeling technique one kind, is from the fluorescence immune body technology, one kind of sensitivity which isotope the immunity technological development comes, special, fast, and can automate modern technology. the enzyme signs method basic principle is unites the antigen either the immune body and the enzyme with the rubber the medicinal preparation union for the enzyme sign antigen or the immune body, this enzyme sign antigen either the immune body may should the antigen or the immune body has the idiosyncrasy with the solid phase carrier on or the organization internal phase, and unified forms still maintained reliably active the immunity compound. When joins the corresponding substrate, the substrate is presented by the enzyme catalysis the corresponding response color. The color depth and corresponding antigen or the immune body content is proportional.Because this technology is the establishment in the antigen - immune body response and in the enzyme highly effective catalytic effect foundation, therefore, has high spirit sensitive and the specificity, is one kind of extremely rich vitality immunology experiment technology.In recent years, the enzyme sign meters was getting more and more popular in clinical laboratory application, caused the enzyme immunity analytic method (EIA) the automaticity and the accuracy is getting higher and higher, particularly in recent years, import, domestically produced list, the multichannel completely automatic enzyme sign meters type and the model development were swift and violent, was the clinical laboratory automaticity after the biochemistry analyzer, the blood corpuscle calculating instrument, the blood congeals the meter and so on to renew once again and to enhance. However, the domestic and foreign related enzyme sign meter performance systems assessments method are really few, some evaluating indicator is simple, and is not comprehensive, some method which uses to its performance evaluation is inconsistent, causes between between the different instrument, the factory and the user, the user and users evaluating indicator lacks the commeasurability, this is because the enzyme sign meter in the fabrication technology (multichannel detector), the determination principle (vertical path of rays photometric method) (721 spectrophotometers) between has the very big difference with other horizontal path of rays luminosity determinations instrument. Therefore, we to the domestic and foreign several different factories and the model instrument after the system research proof, have established a set of more perfect enzyme sign meter performance evaluation target and the appraisal essential method initially. After the preliminary application, the effect is satisfied, should the reading public and users request, plans the enzyme sign meter performance evaluation and the appraisal method theory and the principle does an introduction and the supplement, supplies the colleague when the appraisal, the appraisal and the instrumentation refers, thus to further enhance the test result between the indoor duplication and the room the commeasurability provides the reliable guarantee.The light filter wave length curacy checking and the peak value determine:Method and weight standard: (Wave length precision 0.3 nm) carries on the spectrum scanning with the high accuracy ultraviolet obvious spectrophotometer to the different wave lengths light filter, difference of the examination value and the demarcation value namely for the light filter wave length precision, its differential value more approaches in zero and the peak value more loudly expressed that light filters quality is better, the wave length precision is higher. Rationale: Enzyme sign meters light filter quality quality, immediate influence instruments sensitivity height, but light filters quality is also weighs by its wave length precision and the peak value target, therefore the light filter wave length precision and the peak value weigh one of enzyme sign meters key parameters, although this has not mentioned in the factory instrument instruction booklet, but in instruments actual use process, we discovered that carries on the inspection to the light filter wave length precision and the peak value is important is also necessary, through the inspection may discover that light filters wave length demarcation value and the actual value conform to the degree, may discover whether light filters quality does meet the requirement.Sensitivity and accuracy monitor:Sensitivity: Compounds 6 ug/ml potassium bichromate (dry) precisely the solution (0.05 mo1/L sulfuric acid dissolution), joins 200 ul potassium bichromate solution in the eyelet cup, makes the blank by 0.05 mo1/L sulfuric acid solution, (reference wave length 650 nm) determines in 450 nm, its extinction should = 0.01 A.Accuracy: Accurate configuration: 1mmo/L (raises sterling) to the nitrophenol the peroxide solution, then dilutes it by 10 mmo1/L caustic soda solution 25 times, joins 200 ul diluent in the eyelet cup, solution makes the blank by 10 mmo1/L the NaOH, (reference wave length 650 nm) examines in 405 nm, its extinction should in 0.4 A (0.3950.408 A) aboutExplained: Instruments sensitivity and accuracy mainly two factor influences: First, light filter wave length precision; second, detectors quality. Usually in the situation, the light filter reason causes instruments sensitivity and the accuracy drops quite easily to inspect; But the detector quality difference not easily was discovered that therefore we use the above two standard substance solution to carry on the regular monitor to the instrument detectors quality, thus has guaranteed the instrument test result reliability. Regarding the instrument accurate determination, we have used the literature report method, passes through carries on many times in the different model instrument between determines repeatedly, finally discovered that this standard solution has a more constant determination value in the 450nm wave length place, because enzyme sign meter with other spectrophotometers optics principle not identical, therefore whether can also wait for the colleague as the appraisal enzyme sign meter accurate reference method further studying the researchChannel difference and panel difference examination:Method and turn of expression:Channel difference examination: Takes an enzyme sign board eyelet cup (cup bottom to smooth, transparent, not have scratch, not have pollution) to make the carrier by the enzyme sign plate girder, joins three kind of different densities methyl orange solution 200 ul to put in 8 channels relevant positions successively separately, the distilled water zero alignment, uses the double wave length (determination wave length 490nm, reference wave length 630 or 650nm, the following same) continuously measures three times, observes between its different channel the measurement result uniform available differential value to express extremely its channel is bad.Panel bad survey: Chooses the identical factory, the identical batch number enzyme sign plank (8 altogether 96) to join 200 ul methyl orange solution (extinction to transfer to 0.065 0.070 A) separately to put in the identical channel successively, the distilled water zero alignment, uses the double wave length examination, its erroneous size with 1.96s weighs.Theory explanation: In order to raise the enzyme sign meters examination speed, according to 96 enzyme sign boards specification characteristic, the majority factories middle-grade, the upscale enzyme sign meter used 8 channels detectors (part middle-grade, low-grade enzyme sign meter still to use single channel at present) at present. Therefore their each channels examination ability is different, it is the instrument interior inherent error, with measured that the solution concentration becomes is being related (different detector to different density solution response ability different), therefore the channel difference weighs the enzyme sign meter performance is good or not one of important targets; Its weight target indicated extremely with the difference that this with the factory recommendations target is consistent; The differential value more approaches extremely in zero, the channel difference is smaller, the showing identical sample is better in the different channel check test results uniformity.Because the different factory, between the different batch number enzyme sign boards quality difference, causes the hole with hole between test result not completely consistent, this with horizontal path of rays luxmeter color dish quality whether to meet the requirement to be the same than, therefore we thought that the panel difference is outside instruments inherent error, only then measured accurately knows the panel to be bad, and makes the corresponding adjustment to the result, can cause the check result to tally with the actual situation, more accurate reliable; Uses the evaluating indicator (gentleman 1.96 s) is also is advantageous to the result adjustment. Moreover, in the design panel difference surveys operating procedure, we transfers to 200 ul methyl orange solution extinction 0.0650.070 A, is considered fully adds the type error (Shanghai Refinement Company, 200 ul, gentleman 2%CV), its goal is makes to add the type error control in instruments resolution (0.01 A) below, like this also avoided the panel difference result because of not adding the type error to cause the pseudo markupZero spot drifts:Method: Takes eight eyelet cups, puts in eight channels relevant positions separately, joins 200 ul distilled water and the zero alignment, uses the double wave length or the single wave length (490 nm) every other 30 min determines one time, observes in 8 channel 4h the extinction change. It is zero drifts with a zero differential value.Survey principle: The enzyme sign meters zero spot drifts usually is very small, this is because instrument before the reading, must make 8 channels sample-out counts first (Io). Then reads the sample board again various determination value (I), according to Beer s law light absorption value =1ogl0 (Io/I), each time the reading undergoes the like this automatic correction, causes the error in reading to reduce greatly, such measured that reads the way is without doubt very correct; Moreover some instruments are the application DRL DYE examine the test strip to carry on the absolute extinction the calibration, therefore when uses the single wave length survey, will cause the extinction the pseudo markup, this kind of instrument may adopt two methods to carry on the adjustment; first, chooses appropriate reference light filter to carry on the double wave length determination; second, the introduction revision parameter, namely the single wave length reads 1 blank hole under the extinction pattern, its test value and the predicted value differential value of namely for revision parameter. Therefore zero spot drifts is appraises an instrument withing certain amount of time zero extinction the change tendency, has nothing to do with the wave length, it had reflected indirectly the instrument interior examination system in unit of time is under the active status the stability and instruments mechanical property situation (enters board, examination, withdrawal continuously), therefore it appraises the instrument internal circuit system, the optical system (detector) and the mechanical system is good or not the target.Accuracy appraisal:Method and evaluating indicator: Each channel three small cups, join 200 ul high, middle and low three kind of different densities separately the methyl orange solution, the distilled water zero alignment, uses the double wave length to make the duplicate parallel determination, determines every day two times, determines continuously for 20 days. In distinction computation its batch in accuracy, date batch of accuracy, during the day accuracy and total accuracy and corresponding CV value.Theory basis: in 1984 US Clinical Laboratory scale Committee has published EP5 a T document and has carried on the second revision in 1992: Clinical chemistry instrument precision performance user appraisal. This appraisal plan in instruments and in the reagent and so on biochemistry analyzer, blood corpuscle calculating instrument appraisal obtained the widespread application, caused to appraise instruments method to obtain the unification; But this law applies in the enzyme sign meters appraisal in domestic and foreign not yet sees has the similar report, we use this law to carry on the systems assessment to the enzyme sign meter, the result are quite satisfied. Various targets significance is: In approves the accuracy is by 20d in the daily two batch, each batch of two next best difference (i.e. in batch ) after statistics processing obtained result; In Japan a batch of between accuracy is the daily two batch determines difference of the result average value by 20d in (i.e. batch ) after statistics processing the obtained result; During the day the accuracy expresses in 20 d to measure every day the average value the standard deviation was the standard harms, has reflected the daily average value relative dispersion; The total accuracy is to approving, the batch and during the day the accuracy carries on the weighting statistics the result. And the accuracy and the total accuracys application is most widespread by approving, (i.e. is simultaneously redundant with the traditional batch in accuracys computational method to identical specimen in the same day determines compares many times), the former tallies with the laboratory actual situation, has the representation, is also more reasonable; But the total accuracy objectively comprehensively has reflected the laboratory overall variation situation.Linear determination:Method: The precise name takes the methyl orange to compound 5 series densities the solution, uses the double wave length parallel to examine 8 times asks its average value. The computation regression equation, correlation coefficient r and standard error of estimation Sy, x, and expresses the sample 95% measuring range with 1.96 Sy, x. The evaluating indicator explained: The linear determination also appraises one
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