DZ215單片機控制半自動酶標儀
DZ215單片機控制半自動酶標儀,dz215,單片機,控制,節(jié)制,半自動,酶標儀
單片機控制半自動酶標儀,指導(dǎo)教師:李 巖
學(xué) 生:郭一男
班 級:040301班,題目的來源,近二十年來,由于生物學(xué),近代物理學(xué),化學(xué),電子技術(shù),生物材料學(xué),機械學(xué),計算機等多種科學(xué)的飛速發(fā)展,并愈來愈深入得對生物學(xué)和臨床醫(yī)學(xué)領(lǐng)域的廣泛滲透,促進了醫(yī)學(xué)檢驗理論急實驗室儀器和技術(shù)的發(fā)展.自動化的檢測儀器的涌現(xiàn),大大提高了醫(yī)學(xué)檢驗的測量精度,速度,準確度,從而使繁瑣的操作得以簡化,復(fù)雜的成分及微含量的測量得以實現(xiàn),酶標分析儀就是在這種形式下出現(xiàn)的臨床檢測儀器,酶標儀的檢測原理,酶是生物體內(nèi)細胞產(chǎn)生的具有高效特異催化作用的蛋白質(zhì),有趣的是,一種酶只對某一種化學(xué)反應(yīng)起催化作用,而對其他化學(xué)反應(yīng)不起作用。這就是酶的標記原理。酶聯(lián)免疫吸附測試正是利用了抗體與抗原反應(yīng)的特異性、酶的標記原理和高效催化作用,所以具有高度的特異性和靈敏性。,主要技術(shù)指標,檢測范圍： 0-3A(405nm)；
精度： 讀取范圍 0-4.0A；
準確性 ±1%；
液晶顯示，聲光報警；
打印數(shù)據(jù)，完成與上位機通信。,酶標儀硬件系統(tǒng),酶標儀系統(tǒng)主要包括一下幾個部分：
濾波片
機械操作系統(tǒng)
數(shù)據(jù)的采集與傳輸
單片機控制系統(tǒng)
工作過程如下:
由光電檢測器檢測信號,經(jīng)過放大,A/D轉(zhuǎn)換,送入主機進行數(shù)據(jù)處理,判斷分析,另外由單片機控制電機來實現(xiàn)微面板的運動,系統(tǒng)的總體結(jié)構(gòu),系統(tǒng)框圖:,電機控制,控制系統(tǒng)的選擇,控制芯片選擇單片機89C51:
89C51是一種帶4K字節(jié)閃爍可編程可擦除只讀存儲器的單片機。單片機的可擦除只讀存儲器可以反復(fù)擦除100次。該器件采用ATMEL高密度非易失存儲器制造技術(shù)制造，與工業(yè)標準的MCS-51指令集和輸出管腳相兼容。由于將多功能 8位CPU和閃爍存儲器組合在單個芯片中，ATMEL的89C51是一種高效微控制器，89C51單片機為很多嵌入式控制系統(tǒng)提供了一種靈活性高且價廉的方案。,主要特征:
與MCS-51 兼容，4K字節(jié)可編程閃爍存儲器，壽命100寫/擦循環(huán)，數(shù)據(jù)保留時間：10年，全靜態(tài)工作：0Hz-24Hz，三級程序存儲器鎖定 128*8位，內(nèi)部RAM，32可編程I/O線，兩個16位定時器/計數(shù)器，5個中斷源，可編程串行通道，低功耗的閑置和掉電模式，片內(nèi)振蕩器和時鐘電路。,電機控制系統(tǒng),在控制系統(tǒng)中，選用TA8435H步進電機驅(qū)動芯片的執(zhí)行機構(gòu)，通過步進電機的運動來完成微面板的直線往復(fù)運動。
芯片特點:
工作電壓范圍寬（10V～40V）
輸出電流可達平均為1.5A，峰值可以達到2.5A；
具有整步、半步、細分、細分運行方式可供選擇；
采用脈寬調(diào)制式斬波驅(qū)動方式；
具有正／反轉(zhuǎn)控制功能；
帶有復(fù)位和使能引腳；
可選擇使用單時鐘輸入或雙時鐘輸入；,電路用一片TA8435H來驅(qū)動電機，輸入信號有使能控制、正反轉(zhuǎn)控制和時鐘輸入，通過光耦可將驅(qū)動器與輸入級進行電隔離，以起到邏輯電平隔離和保護作用；該電路工作在1/8細分模式（M1、M1接高電平），可減小低速時的振動.
另外采用外部定時／計數(shù)器82C53來給TA8435H提供步進脈沖。
82C53的工作方式是一種方波速率發(fā)生器。在這種方式下，當CPU設(shè)置控制好后，輸出將為高電平，在寫完計數(shù)值后就自動開始計數(shù)，輸出保持高電平；而當計到一半計數(shù)值時，輸出變低直到計數(shù)到0，此后輸出又變高以重新開始計數(shù)。,放大電路系統(tǒng)設(shè)計,系統(tǒng)設(shè)計要求的檢測范圍是0～405nm,檢測到的電流應(yīng)在0～100nA,通過一個5K電阻得到的電壓值在0～0.0005V，在經(jīng)過兩級放大之后，由于一級放大的倍數(shù)為100倍，那兩級放大之后，使其電壓值可以達到0～5V，剛好符合A/D轉(zhuǎn)換芯片AD976A輸入電壓的范圍，符合系統(tǒng)的設(shè)計要求。,,,,A/D轉(zhuǎn)換電路設(shè)計,在A/D轉(zhuǎn)換電路中,選用由AD公司生產(chǎn)的AD976A, AD976A是AD公司推出的l6位高速,高精度A/D轉(zhuǎn)換器,最大通過率為100kSPS，輸入信號范圍為-l0一+I0V,帶寬為1.5MHz，它可以與8位,16位單片機和DSP方便接口,使用十分方便靈活。
它的主要特征如下:
(1)AD976A是具有100kSPs／200kSPs高通過率的并行輸出ADC；
(2)需+5V單電源供電，且具有100mW的低功耗：
(3)片上集成有ADC、參考電源和時鐘；
(4)采用開關(guān)電容／電荷重分布結(jié)構(gòu)，其內(nèi)部的自動校正邏輯可以校正內(nèi)部的非線性，從而使其性能總體上得到了優(yōu)化。,A/D轉(zhuǎn)換電路硬件連接圖:,顯示與鍵盤系統(tǒng)設(shè)計,顯示系統(tǒng):
酶標儀具有顯示功能，本設(shè)計中，主要顯示的是各路采集的數(shù)據(jù)，同時顯示時間日期等功能，所以用一般的LED顯示可能很難實現(xiàn)，所以我采用LCD液晶顯示，并采用PCF2111CT字符液晶驅(qū)動模塊。,PCF2111CT模塊的主要特點有：
（1）工作電壓要求范圍大，DC2.25V—6.0V；低功耗，最大功耗100mW；
（2）內(nèi)置振蕩電路，無需外接振蕩器，只要在振蕩器輸入端與電源跨接一個電容，與地跨接一個電阻即可，使用設(shè)計方便；
（3）數(shù)據(jù)串行連續(xù)輸入，與外圍電路特別是單片機連接時，只需3根線，可以節(jié)約單片機的I/O口
（4）具有加電自動復(fù)位功能；
（5）具有液晶底板背光驅(qū)動功能。,液晶顯示電路連接圖:
39腳、40腳分別接單片機，37、38腳接發(fā)光二極管，用于液晶的背光顯示。單片機的P1.0、P1.1、P1.2三腳分別充當同步寫入時鐘CLB、數(shù)據(jù)輸入DATA、輸入控制DLEN的功能。初始化單片機時置P1.2為低電平，寫入數(shù)據(jù)時，置P1.2高電平。,鍵盤系統(tǒng):
系統(tǒng)的鍵盤輸入主要有0到8路數(shù)據(jù)采集的控制，復(fù)位等功能，需要的鍵盤數(shù)量較多，我采用8×2矩陣鍵盤，控制芯片則采用HD7279A。
HD7279A是一種管理鍵盤和LED顯示器的專用智能控制芯片，同時能對多達8×8的鍵盤矩陣的按鍵情況進行監(jiān)視，具有自動消除鍵抖動并識別按鍵代碼的功能，從而可以提高CPU工作的效率.
HD7279A和微處理器之間采用串行接口，其接口電路和外圍電路簡單，占用口線少，加之它具有較高的性能價格比，因此，在微型控制器、智能儀表、控制面板和家用電器等領(lǐng)域中日益獲得廣泛的應(yīng)用。,軟件設(shè)計,在主程序中，首先對單片機80C51等
功能芯片進行初始化，
設(shè)定各項初值。然后檢測按鍵，
進行按鍵處理。鍵盤處理結(jié)束后，
通過電機運動，
實現(xiàn)數(shù)據(jù)采集。,因為需要采集8路數(shù)據(jù)，所以用ch表示所采集的路段，其采集子程序流程圖如圖,謝謝各位老師指導(dǎo), 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 meter's photoelectric colorimeter.First introduced the enzyme sign meter's 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 direction's 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 fluid's 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 sample's plastic micro orifice is the multi-platoons porous, the optical fiber can only vertical pass through, therefore the enzyme sign meter's 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 meter's 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 channel's instruments are equipped with 12 light beams or 12 photo sources, 12 detectors and 12 amplifiers, under X direction's machinery drive's function, sample 12 are one row are examined. The multichannel enzyme sign meter's 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 sign's 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 meter's 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 meter's 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 assessment's 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 user's 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 determination's 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 user's 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 length's 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 filter's quality is better, the wave length precision is higher. Rationale: Enzyme sign meter's light filter quality quality, immediate influence instrument's sensitivity height, but light filter's 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 meter's key parameters, although this has not mentioned in the factory instrument instruction booklet, but in instrument's 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 filter's wave length demarcation value and the actual value conform to the degree, may discover whether light filter's quality does meet the requirement.Sensitivity and accuracy monitor:①Sensitivity: 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.②Accuracy: 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 (0.395~0.408 A) aboutExplained: Instrument's sensitivity and accuracy mainly two factor influences: First, light filter wave length precision; second, detector's quality. Usually in the situation, the light filter reason causes instrument's 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 detector's 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 spectrophotometer's 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:①Channel 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 channel's 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.②Panel 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 meter's examination speed, according to 96 enzyme sign board's specification characteristic, the majority factories middle-grade, the upscale enzyme sign meter used 8 channel's detectors (part middle-grade, low-grade enzyme sign meter still to use single channel at present) at present. Therefore their each channel's 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 recommendation's 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 result's uniformity.Because the different factory, between the different batch number enzyme sign board's 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 instrument's 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 survey's operating procedure, we transfers to 200 ul methyl orange solution extinction 0.065~0.070 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 instrument's 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 channel's 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 meter's zero spot drifts usually is very small, this is because instrument before the reading, must make 8 channel's 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 instrument's 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 instrument's method to obtain the unification; But this law applies in the enzyme sign meter's 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 accuracy's application is most widespread by approving, (i.e. is simultaneously redundant with the traditional batch in accuracy's 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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