Quawell微量紫外分光光度計.ppt

1、微量分光光度計,型號：Q3000 Q5000 品牌：,目錄,一.概述 二.Q5000使用詳解,一.概述,Q3000使用高能量LED燈提供260nm、280nm的光譜檢測，不需要暖機，開機即可使用，配有簡易操作界面，以及硅電二極管檢測器，不需用比色杯，在5秒鐘內(nèi)就可以完成對0.5-2.5ul的樣品的測量，檢測吸收值可達80Abs(dsDNA濃度 0-4000 ng/ul)，大部分純化后的生物大分子幾乎都不需要稀釋、濃縮就可以上機檢測。,Q5000使用高能量氙燈提供200-850nm的全光譜檢測，不需要暖機，開機即可使用。搭配高感度的CCD- array檢測器，檢測吸收值可高達100Abs (ds

2、DNA濃度1ng/ul-5000ng/ul)大部分純化后的生物大分子幾乎都不需要稀釋濃縮即可上機檢測。不需用比色杯，在5秒鐘內(nèi)就可以完成對0.5-2.5ul的樣品的測量。,產(chǎn)品用途,1.核酸濃度檢測：檢測dsDNA、ssDNA、RNA等核酸濃度； 2.探針檢測：檢測熒光標記探針的吸光度值，可用于去除未能標記探針的樣品； 3.蛋白濃度檢測：檢測普通純化后蛋白質(zhì)的濃度； 4.蛋白標記檢測：可檢測被BCA、Bradford或Lowry精確定量待測蛋白濃度，自動畫出標準曲線并計算出濃度； 5.糖類、醛類等生物大分子檢測：常規(guī)紫外光波下檢測樣品吸光值； 6.細胞培養(yǎng)物檢測：檢測細胞培養(yǎng)物在600nm處的

3、Abs。（Q5000）,工作原理,應(yīng)用液體的表面張力特性，只需要0.5-2.5ul的液體樣品，便能在上下檢測臂之間拉出一個標準的小液柱，儀器檢測通過小液柱的光線的變化情況，得到溶液的吸光值，進而計算出所含溶質(zhì)濃度。,產(chǎn)品特點,樣品體積?。?.5-2.5ul的樣品就可以完成檢測； 檢測濃度范圍寬：無需濃縮或稀釋樣品就可以檢測； 操作簡單：無需比色杯，減少了反復(fù)清洗比色杯帶來的麻煩，減少了實驗誤差； 檢測方便快捷：不需要預(yù)熱，開機就可使用； 數(shù)據(jù)處理簡便：操作界面便捷，數(shù)據(jù)可輕松轉(zhuǎn)存到Excel中。,技術(shù)參數(shù),1. 樣品量：0.5-2.5ul 2.波長范圍：200-850nm / 260nm、28

4、0nm、380nm (Q5000/Q3000 下同) 3.波長精度：1nm 4.分辨率：1nm 5.其他：1mm光程長度（可自動調(diào)整到0.05mm） 6.檢測下限：1ng/ul 7.檢測上限：5000ng/ul / 4000ng/ul 8.吸光率精確度：0.002 Abs(1mm光程) 9.吸光率準確性：1%（at 0.76 at 257nm） 10.吸光率范圍：0.01-100Abs / 0-80Abs 11.檢測器類型：3648個單元硅CCD陣檢測器/硅電二極管檢測器 12.核酸檢測周期：8s/ 2s 13.光源：氙燈/LED 14.軟件兼容性：Windows XP ,Vista(32 b

5、it) 15.操作電壓：12V 16.運作功率：15W 17.待機功率：1.5W /5W 18.通過CE認證,注意事項,1.對于一般的核酸、蛋白質(zhì)樣品，檢測前徐使用漩渦振蕩器震蕩均勻為最佳，或至少以移液 器吸放數(shù)次混勻。若擔(dān)心DNA可能因前述動作而斷裂，可改以55加熱約一分鐘，使樣品 在檢測前呈均勻狀態(tài)，以確保用于上樣檢測的2ul樣品具有代表性。 2.檢測后應(yīng)當(dāng)立即用拭鏡紙擦凈上下檢測孔。 3.同一滴液體只能做一次檢測，欲重復(fù)定量同一樣品，請擦掉前一滴，重新取出一滴進行 檢測。 4.核酸上樣為1ul-2ul，蛋白為2ul，上樣需一次完成。 5.大部分檢測錯誤源于未成形正確的液柱，正確液柱如下圖

6、所示。如若未出現(xiàn)正確液柱，應(yīng)當(dāng)擦掉液滴，重新加樣。 6.不能使用含有腐蝕性的液體。,定期維護,儀器使用一段時間后，可能出現(xiàn)檢測孔污染，這時候儀器會自檢并提醒維護，可用0.5的次氯酸鈉和無水乙醇進行清洗，方法如下： 1.使用去離子水潤洗30秒，擦鏡紙擦干； 2.使用漂白劑氧化去污，通常0.5次氯酸鈉即可，半分鐘后擦鏡紙擦干； 3.使用無水乙醇清洗30s左右，半分鐘后擦鏡紙擦干； 4.重復(fù)步驟1，擦干凈后將儀器放在無塵環(huán)境中。,二.Q5000使用詳解,軟件安裝,插入光盤，雙擊“Setup”文件，按照提示完成軟件安裝。 使用USB數(shù)據(jù)線將儀器與電腦相連，驅(qū)動將自動安裝。 注意：在軟件安裝前不要將Q5

7、000與電腦相連。 當(dāng)在Vista或者Win 7中安裝軟件時，若出現(xiàn)“版本沖突”對話框，請選擇選項“是”。 若出現(xiàn)對話框有“放棄”、“重試”、“忽略”三個選項時，請選擇選項“忽略”，使安裝繼續(xù)進行。 點擊主界面下的獲得軟件升級或產(chǎn)品新聞。,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 u

8、ser to adjust the light integration. 2. Dye List: edit the dye data,APPLICATION MODULE INTERFACE,1. Checkbox 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 bei

9、ng measured. 3. Main function Panel: contains all main functions for sample measuring and data handling. 4. Graph area: 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

10、單中選擇測量模式； 2. 選擇樣品類型； 3. 輸入樣品名稱； 4. 空白對照: 打開上測量臂，將2.5 ul空白對照溶液加到基座上，放下上臂，點擊 “Blank” 按鈕；結(jié)束后打開上臂，用擦鏡紙拭去上下表面的液體 ； 5. 吸取2.5 ul待測液加到基座上，點擊 “Measure” 按鈕進行測量。,Checking the Blanking Result,建議把空白對照溶液當(dāng)做待測液進行測量，在10mm的吸光值結(jié)果應(yīng)該不大于0.04，如果不是，請重新空白對照并檢測。 原文：It is recommended that measuring the blanking buffer as if i

11、t 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 more than 0.04 at 10 mm absorbance equivalent, if not, repeat the 1 and 2.,使用圖示,SAVING THE DATA AND INFORMATION,Note: Q5000 program does not save the dat

12、a 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 to export the data from the data table to an Excel spreadsheet.,Example1：NUCLEIC ACIDS,Sample Volume There is no specific requirement for the sa

13、mple volume; however for the best accuracy and reproducibility we recommend 2-2.5ul. SW and Abs (10mm): user-selected wavelength and corresponding absorbance that are not utilized in any calculations. Sample Type EC: dsDNA, ssDNA and RNA.,Example2：MICROARRAY,The Microarray module analyzes fluorescen

14、tly-labeled nucleic acid probes. It simultaneously measures the concentration of the fluorescent tag and the nucleic acid at appropriate wavelengths. 微序列模式用來分析熒光標記核酸探針。它同時測量在合適的波長處的熒光標記和核酸的濃度。,Sample Type EC: dsDNA, ssDNA and RNA. Dye1 and Dye 2 Selection Windows 1. Dye1 or Dye2 drop-down list: disp

15、lays 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: concentration of Dye1 or Dye2 in pmol/nl. 4. Vertical Lines: the green line indicates the peak position of the wavelength for

16、 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 calculates the concentration. Since no protein standards are required, Protein A280 is fast and convenient. Protein A280測量蛋白質(zhì)

17、在280nm處的吸光值并計算其濃度。因為不需要標準曲線，所以檢測快速簡便。 Sample Type: 1Abs =1mg/ml, BSA, IgG, and Lysozyme.,Example4：LABELED-PROTEINS,The Labeled-Proteins function will simultaneously measure both protein and fluorescent dye concentrations at appropriate wavelengths.,Sample Type: 1Abs =1mg/ml, BSA, IgG, and Lysozyme D

18、ye1 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 in uM. 4. Vertical Lines: the green line on the absorbance-wavelength graph indicates the peak p

19、osition 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 spectrophotometer. The UV-Vis measurement module provides the operator with a sample absorbance rang

20、e from 220 to 850 nm. Q5000 是一款功能強大的實驗室分光光度計。 UV-Vis measurement 模式可以在220-850nm的波長范圍內(nèi)對樣品的吸光值進行測量。,(1) nm: select the wavelength by using the up/down arrows. Abs 1 (1mm): corresponding absorbance at (1) nm. (2) nm: select the wavelength by using the up/down arrows. Abs 2 (1mm): corresponding absorban

21、ce at (2) nm. 1 和2設(shè)定需要測定吸光值的波長位置。 Baseline: if selected, the absorbance value of the baseline is subtracted from the absorbance of Abs1 and Abs2. 如果選擇此項，則基準線處吸光值將從Abs1 和Abs2中去除。 Vertical Lines: the green line on the absorbance-wavelength graph indicates the peak position of the wavelength for (1)nm,

22、 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 at 600 nm. Q5000 可以通過檢測600nm處的光散射情況來觀察懸浮細胞或微生物培養(yǎng)物的密度。,Baseline:

23、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 selected nm.,THE PREDEFINED FLUORESCENT DYES LIST,The list w

24、orks 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 the light integration. The integration number is se

25、t 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, the light integration can be adjusted with this

26、 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 value (1-12)”. 4. Click the “Check” an

27、d 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 OF ERROR,樣品殘留（Sample Overlap ）

28、 解決辦法： 1.每次測量后用干的擦鏡紙擦凈上下檢測孔表面； 2.測完高濃度樣品后用2ul去離子水清洗上下表面； 3.全部測量完畢后，用去離子水清洗上下表面及周圍； 4.樣品測量前按下面方法空白儀器（ blank the machine）： 1. 打開適合的應(yīng)用模式. 2. 吸取2.5 ul空白溶液加到基座上. 3. 放下上臂，點擊Blank按鈕. 4. 打開上臂，擦干液體并用去離子水清洗. 5.吸取2.5 ul空白溶液加到基座上. 6.放下上臂，點擊Measure按鈕. 7. 如果260nm處吸光值高于0.004, 重復(fù)以上步驟。,樣品同質(zhì)性（Sample Homogeneity） Non-homogenous samples would cause significant deviation in the data generated by any measurement s