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1、微量分光光度计,目录,一.概述二.Q5000使用详解,一.概述,Q3000使用高能量LED灯提供260nm、280nm的光谱检测，不需要暖机，开机即可使用，配有简易操作界面，以及硅电二极管检测器，不需用比色杯，在5秒钟内就可以完成对0.5-2.5ul的样品的测量，检测吸收值可达80Abs(dsDNA浓度 0-4000 ng/ul)，大部分纯化后的生物大分子几乎都不需要稀释、浓缩就可以上机检测。,Q5000使用高能量氙灯提供200-850nm的全光谱检测，不需要暖机，开机即可使用。搭配高感度的CCD-array检测器，检测吸收值可高达100Abs(dsDNA浓度1ng/ul-5000ng/ul)
2、大部分纯化后的生物大分子几乎都不需要稀释浓缩即可上机检测。不需用比色杯，在5秒钟内就可以完成对0.5-2.5ul的样品的测量。,产品用途,1.核酸浓度检测：检测dsDNA、ssDNA、RNA等核酸浓度；2.探针检测：检测荧光标记探针的吸光度值，可用于去除未能标记探针的样品；3.蛋白浓度检测：检测普通纯化后蛋白质的浓度；4.蛋白标记检测：可检测被BCA、Bradford或Lowry精确定量待测蛋白浓度，自动画出标准曲线并计算出浓度；5.糖类、醛类等生物大分子检测：常规紫外光波下检测样品吸光值；6.细胞培养物检测：检测细胞培养物在600nm处的Abs。（Q5000）,工作原理,应用液体的表面张力特
3、性，只需要0.5-2.5ul的液体样品，便能在上下检测臂之间拉出一个标准的小液柱，仪器检测通过小液柱的光线的变化情况，得到溶液的吸光值，进而计算出所含溶质浓度。,产品特点,样品体积小：的样品就可以完成检测；检测浓度范围宽：无需浓缩或稀释样品就可以检测；操作简单：无需比色杯，减少了反复清洗比色杯带来的麻烦，减少了实验误差；检测方便快捷：不需要预热，开机就可使用；数据处理简便：操作界面便捷，数据可轻松转存到Excel中。,技术参数,1.样品量：2.波长范围：200-850nm/260nm、280nm、380nm(Q5000/Q3000 下同)3.波长精度：1nm4.分辨率：1nm5.其他：1mm光
4、程长度（可自动调整到0.05mm）6.检测下限：1ng/ul7.检测上限：5000ng/ul/4000ng/ul8.吸光率精确度：0.002 Abs(1mm光程)9.吸光率准确性：1%（at 0.76 at 257nm）10.吸光率范围：0.01-100Abs/0-80Abs11.检测器类型：3648个单元硅CCD阵检测器/硅电二极管检测器12.核酸检测周期：8s/2s13.光源：氙灯/LED14.软件兼容性：Windows XP,Vista(32 bit)15.操作电压：12V16.运作功率：15W17.待机功率：1.5W/5W18.通过CE认证,注意事项,1.对于一般的核酸、蛋白质样品，检
5、测前徐使用漩涡振荡器震荡均匀为最佳，或至少以移液 器吸放数次混匀。若担心DNA可能因前述动作而断裂，可改以55加热约一分钟，使样品 在检测前呈均匀状态，以确保用于上样检测的2ul样品具有代表性。2.检测后应当立即用拭镜纸擦净上下检测孔。3.同一滴液体只能做一次检测，欲重复定量同一样品，请擦掉前一滴，重新取出一滴进行 检测。4.核酸上样为1ul-2ul，蛋白为2ul，上样需一次完成。5.大部分检测错误源于未成形正确的液柱，正确液柱如下图所示。如若未出现正确液柱，应当擦掉液滴，重新加样。6.不能使用含有腐蚀性的液体。,定期维护,仪器使用一段时间后，可能出现检测孔污染，这时候仪器会自检并提醒维护，可
6、用0.5的次氯酸钠和无水乙醇进行清洗，方法如下：1.使用去离子水润洗30秒，擦镜纸擦干；2.使用漂白剂氧化去污，通常0.5次氯酸钠即可，半分钟后擦镜纸擦干；3.使用无水乙醇清洗30s左右，半分钟后擦镜纸擦干；4.重复步骤1，擦干净后将仪器放在无尘环境中。,二.Q5000使用详解,软件安装,插入光盘，双击“Setup”文件，按照提示完成软件安装。使用USB数据线将仪器与电脑相连，驱动将自动安装。注意：在软件安装前不要将Q5000与电脑相连。当在Vista或者Win 7中安装软件时，若出现“版本冲突”对话框，请选择选项“是”。若出现对话框有“放弃”、“重试”、“忽略”三个选项时，请选择选项“忽略”
7、，使安装继续进行。点击主界面下的获得软件升级或产品新闻。,MAIN MENU,The Measurement tab:1.Nucleic Acids 2.Microarray 3.UV-Vis Measure 4.Protein A280 5.Labeled Protein 6.Cell Culture The Tools tab:1.Light Integration:Allows the user to adjust the light integration.2.Dye List:edit the dye data,APPLICATION MODULE INTERFACE,1.Check
8、box Panel:contains online-help and other options.The user can select the option based on the particular need.2.Current data display windows:display the data and information of the sample being measured.3.Main function Panel:contains all main functions for sample measuring and data handling.4.Graph a
9、rea:displays the absorbance(Y-axis)and spectrum(X-axis).5.Data Table:used for holding and displaying the accumulated data.The maximum number of rows is 1000.,Measurement,1.在主菜单中选择测量模式；2.选择样品类型；3.输入样品名称；4.空白对照:打开上测量臂，将2.5 ul空白对照溶液加到基座上，放下上臂，点击“Blank”按钮；结束后打开上臂，用擦镜纸拭去上下表面的液体；5.吸取2.5 ul待测液加到基座上，点击“Meas
10、ure”按钮进行测量。,Checking the Blanking Result,建议把空白对照溶液当做待测液进行测量，在10mm的吸光值结果应该不大于0.04，如果不是，请重新空白对照并检测。原文：It is recommended that measuring the blanking buffer as if it were a sample:1.Establish a blank as above“4”.2.Measure the blanking buffer as if it were a sample as above“5”.3.The result should be no m
11、ore than 0.04 at 10 mm absorbance equivalent,if not,repeat the 1 and 2.,使用图示,SAVING THE DATA AND INFORMATION,Note:Q5000 program does not save the data automatically.All data and information for the sample being measured will be lost if it is not saved.User can save the data using the“Report”function
12、 to export the data from the data table to an Excel spreadsheet.,Example1：NUCLEIC ACIDS,Sample Volume There is no specific requirement for the sample volume;however for the best accuracy and reproducibility we recommend 2-2.5ul.SW and Abs(10mm):user-selected wavelength and corresponding absorbance t
13、hat are not utilized in any calculations.Sample Type EC:dsDNA,ssDNA and RNA.,Example2：MICROARRAY,The Microarray module analyzes fluorescently-labeled nucleic acid probes.It simultaneously measures the concentration of the fluorescent tag and the nucleic acid at appropriate wavelengths.微序列模式用来分析荧光标记核
14、酸探针。它同时测量在合适的波长处的荧光标记和核酸的浓度。,Sample Type EC:dsDNA,ssDNA and RNA.Dye1 and Dye 2 Selection Windows 1.Dye1 or Dye2 drop-down list:displays the dye that is pre-predefined using the Dye List Editor.Please see Section 11 for details on how to predefine the list.2.Abs:absorbance of Dye1 or Dye2.3.pmol/nl:c
15、oncentration of Dye1 or Dye2 in pmol/nl.4.Vertical Lines:the green line indicates the peak position of the wavelength for Dye 1,and the blue vertical line indicates the peak position of the wavelength for Dye 2.,Example3：PROTEIN A280,Protein A280 measures the proteins absorbance at 280 nm and calcul
16、ates the concentration.Since no protein standards are required,Protein A280 is fast and convenient.Protein A280测量蛋白质在280nm处的吸光值并计算其浓度。因为不需要标准曲线，所以检测快速简便。Sample Type:1Abs=1mg/ml,BSA,IgG,and Lysozyme.,Example4：LABELED-PROTEINS,The Labeled-Proteins function will simultaneously measure both protein and
17、fluorescent dye concentrations at appropriate wavelengths.,Sample Type:1Abs=1mg/ml,BSA,IgG,and LysozymeDye1 and Dye 2 Selection Windows 1.Dye1 or Dye2 drop-down list:displays the dye that is pre-predefined using the Dye List Editor.2.Abs:absorbance of Dye1 or Dye2.3.uM:concentration of Dye1 or Dye2
18、in uM.4.Vertical Lines:the green line on the absorbance-wavelength graph indicates the peak position of the wavelength for Dye 1,and the blue vertical line indicates the peak position of the wavelength for Dye 2.,Example5:UV-VIS MEASUREMENT,The Q5000 can function as a general-use laboratory spectrop
19、hotometer.The UV-Vis measurement module provides the operator with a sample absorbance range from 220 to 850 nm.Q5000 是一款功能强大的实验室分光光度计。UV-Vis measurement 模式可以在220-850nm的波长范围内对样品的吸光值进行测量。,(1)nm:select the wavelength by using the up/down arrows.Abs 1(1mm):corresponding absorbance at(1)nm.(2)nm:select
20、the wavelength by using the up/down arrows.Abs 2(1mm):corresponding absorbance at(2)nm.1 和2设定需要测定吸光值的波长位置。Baseline:if selected,the absorbance value of the baseline is subtracted from the absorbance of Abs1 and Abs2.如果选择此项，则基准线处吸光值将从Abs1 和Abs2中去除。Vertical Lines:the green line on the absorbance-wavele
21、ngth graph indicates the peak position of the wavelength for(1)nm,and the blue vertical line indicates the peak position of the wavelength for(2)nm.,Example5:CELL CULTURES,The Q5000 allows laboratories to monitor the density of suspended cell and microbial cultures by measuring their light scatter a
22、t 600 nm.Q5000 可以通过检测600nm处的光散射情况来观察悬浮细胞或微生物培养物的密度。,Baseline:if selected,the absorbance value of the baseline is subtracted from the absorbance at 600nm.Selected nm:user select wavelength.Abs(1mm):absorbance at selected nm.Green vertical line:indicates the peak position of the wavelength for selecte
23、d nm.,THE PREDEFINED FLUORESCENT DYES LIST,The list works with both the Microarray and Labeled Proteins modules.This list contains a predefined list of fluorescent dyes as shown below.,Additional fluorescent dyes can be added by the user as needed.,LIGHT INTEGRATION,This module is used for adjusting
24、 the light integration.The integration number is set to normal before shipping.The light integration and detector systems are functioning normally if the red and black spectra appear as in the image below.,However,if the image appears as below,with the light integration spectrum too high or too low,
25、the light integration can be adjusted with this feature.,Adjusting the light integration:1.Select the module from the horizontal tab and the path from the vertical tab.2.The original integration number appears in the window“Original Value”.3.Enter the number in the window“Enter new light integration
26、 value(1-12)”.4.Click the“Check”and the black spectrum will display in the window.Increase the integration value from 1 up to12,stopping when the black and red spectra are close.If you have questions about the light integration,please contact your local distributer or Quawell at.,POTENTIAL SOURCES O
27、F ERROR,样品残留（Sample Overlap）解决办法：1.每次测量后用干的擦镜纸擦净上下检测孔表面；2.测完高浓度样品后用2ul去离子水清洗上下表面；3.全部测量完毕后，用去离子水清洗上下表面及周围；4.样品测量前按下面方法空白仪器（blank the machine）：1.打开适合的应用模式.2.吸取2.5 ul空白溶液加到基座上.3.放下上臂，点击Blank按钮.4.打开上臂，擦干液体并用去离子水清洗.5.吸取2.5 ul空白溶液加到基座上.6.放下上臂，点击Measure按钮.7.如果260nm处吸光值高于0.004,重复以上步骤。,样品同质性（Sample Homogene
28、ity）Non-homogenous samples would cause significant deviation in the data generated by any measurement system,including the spectrophotometer.,蒸发的影响（Effect of Evaporation）Sample evaporation could cause a 1-2%deviation in the samples concentration.,样品体积不足（Insufficient Sample Volume）We recommend 2.5 ul
29、 for normal sample measurements.,TROUBLESHOOTING,USB Connection Error This error screen usually indicates that the Q5000 and PC connection failed.To correct the error,unplug the USB cable,wait for 2-3 seconds,and plug it again.Sample Accuracy and Reproducibility If the sample data are inaccurate and
30、/or not reproducible please refer to“POTENTIAL SOURCES OF ERROR”.,MAINTENANCE,The primary maintenance requirement of the Q5000 spectrophotometer is to keep the measurement surfaces clean.Upon completion of a measurement,wipe the sample from the upper and lower surfaces.Clean the surfaces and surrounding area with de-ionized water to prevent sample carryover and residue buildup.日常维护要求保持上下检测表面干净。每次测量任务完成后用去离子水清洗检测面及周围以避免样品残留。,谢谢！,
