SoftWare. M.Wave Professional PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION

Operation Manual SoftWare M.Wave Professional PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION 3, Hagavish st. Israel 58817 Tel: 972 3 5595252, Fax: 972 3 5594529 mrc@mrclab.com MRC.011.14

Contents 1. Functions...1 1.1 Main Functions...1 2. Installation...2 2.1 System Requirements...2 2.2 Install M.Wave Professional...2 2.3 Uninstall M.Wave Professional...2 2.4 Rum M.Wave Professional...3 2.5 Set Communication Port...3 2.6 Set User s Information...3 2.7 Set Data Format...4 2.8 Set Interface Style...4 2.9 Connect to PC...4 3. Introduction...5 3.1 Main Interface...5 3.2 Menu & Tools Bar...5 4. Operation...6 4.1 Quantitative Analysis...6 4.2 Kinetics Analysis...8 4.3 Spectrum Scan...9 4.4 Multi Wavelength Analysis...10 4.5 DNA/Protein Analysis...11 4.6 Energy Scan...12 4.7 Modify a Result...13 4.8 Delete a Result...13 4.9 Name a Result...13 4.10 Spectrum Processing...13 4.10.1 Switch Display Mode...14 4.10.2 Zoom Selected Area...14 4.10.3 Modify Scale...14

4.10.4 Auto Fitting...14 4.10.5 Select as Current Spectrum...14 4.10.6 Change the Color of Current Spectrum...14 4.10.7 Search Peaks...14 4.10.8 Mathematics Functions...15 4.10.9 Spectrum Smooth...15 5. Other Functions...16 5.1 Turn On/Off W Lamp...16 5.2 Turn On/Off D2 Lamp...16 5.3 Set Light Source Switch Point...16 5.4 Set Wavelength...16 5.5 Get Dark Current...16 5.6 Scan System Baseline...16 5.7 Auto 8-Cells Holder...16 5.8 Cell Calibration...16 6. File Operation...18 6.1 Save a Testing...18 6.2 Open a Testing...18 6.3 Print Test Report...18 6.4 Export to Excel...18 6.5 Export Curves as Bmp Picture...18 Appendix 1 Methods of Quantitative Analysis...19 Appendix 2 Methods of DNA/Protein Analysis...19

Functions 1. We ll introduce the main functions of M.Wave Professional in this chapter. 1.1 Main Functions Quantitative Analysis We provide 2 methods to establish a Standard Curve; Up to 20 standard samples to establish a new curve or input the coefficients to make a standard curve; There are three methods, which are which are Linear, Square and Cubic ways to fit the standard curve. Kinetics Analysis We can choose the Time Intervals(0.5, 1.0, 2.0, 5.0, 10.0, 30.0 or 60.0 Seconds); You can choose different Photometric Mode to display the curve (TransmittanceTime & Absorbance-Time). Spectrum Scan You can save and load the System Baseline; You can choose the Scan Intervals(0.1, 0.2, 0.5, 1.0, 2.0 and 5nm); You can choose the Photometric Mode to display the spectrum.(wavelengthtransmittance, Wavelength-Absorbance); Automatically list spectrum peaks; Mathematics and smooth. Multi-wavelength Analysis Up to20 wavelengths to measure a sample. DNA/Protein Analysis We provide 2 methods; You can input the coefficients by yourself. Energy Scan You can switch the light source point(w Lamp, D2 Lamp or automatically switch); You can choose the scanning intervals(0.1,0.2,0.5,1.0,2.0 and 5.0); You can set the gain(1 to 8 times); Automatically search wavelength peak.

2. Installation We will show you how to install M.Wave Professional on your PC in this chapter. 2.1 PC System Requirements Pentium or above PC; CD-ROM; 2 USB Ports. 32MB Memory(256MB or Above is strongly recommended); 50MB or above hard disc Space; Microsoft Windows 2000, Windows XP or Vista. 2.2 Install M.Wave Professional Put M.Wave Professional disc in the CD-ROM; Double Click to open the CD-ROM, and then, double click Setup.exe which is under the root directory of CD to start installation (Fig.2-1), click Next; Input user s information (Fig.2-2), click Next; Choose install path (Fig.2-3), then click Next; Choose Shortcut file folder, then click Next, all the setting information will be displayed on the screen (Fig.2-5), click Next to copy files to Hard disc; Click Finish to finish the installation(fig.2-6). Fig.2-1 Welcome Interface Fig.2-3 Install Path Fig.2-2 User s Information Fig.2-4Choose Shortcut Folder Fig2-5 Display User Information Fig.2-6 Finish Installation 2.3 Uninstall M.Wave Professional Start All Files M.Wave Professional Uninstall M.Wave Professional 2.0 (Fig.2-7), click Next to uninstall. After all the files were removed (Fig.2-8), click Finish to end.

Fig.2-7 Uninstall Information Fig.2-8 Finish Uninstallation 2.4 Run M.Wave Professional Insert the USB Key to the USB Port. (Fig.2-9). Two ways are under your choice to run M.Wave Professional. 1. Double click the icon on desktop. 2. Start All Files M.Wave Professional M.Wave Professional 2.0. Fig.2-9 Install USB Key 2.5 Set up Communication Port Be sure that the USB line which is connecting PC with instrument is working and the spectrophotometer is at the standby state interface. After M.Wave Professional being started, then click Main Menu Option, a dialog box of Option will display in the screen(fig.2-10), click Search to automatically search the instrument s communication port, after get the communication port, click OK to save setting. If you choose the option of Connect to Spectrophotometer When Start, the instrument will connect to the PC automatically when you run M.Wave Professional next time. Fig.2-10 Search the Communication Port 2.6 Set User s Information Click Main Menu Option, pop-up Option windows, click User s Information tag, choose it and input relative user information (Fig.2-11), click OK to save settings, all this information will appear in your testing report.

Fig.2-11 Set User s Information 2.7 Set Data Format Click Main Menu Option, a dialog box of Option will display in the screen, click Data Format tag, choose the displaying format (Fig.2-12), and click OK to save setting. Fig.2-12 Set Data Format 2.8 Set Interface Style Click Main Menu Option, a dialog box of Option will display in the screen, and click Interface tag. You can custom the style and color schemes of the interface as you like. Click OK to save setting. Fig.2-13 Set Interface Style 2.9 Connect to PC After enter the Main Interface, click in the Tool Bar to connect with PC. After connection, the icon is on the pitched on state, you can release PC if you click the icon again.

3. Introduction We will introduce the IDE of M.Wave Professional in this chapter. 3.1 Main Interface This is the main interface after start. (Fig.3-1) Fig.3-1 Main Interface 3.2 Menus Bar and Tools Bar Menus Bar and Tools Bar provide three different ways to operate this software. You can finish all the operation through Keyboard or Mouse. Most of the functions can be finished through the Shortcut Bar. (Fig.3-3) Most of the functions can be finished through the menu by Right-hand button. (Fig.3-4) Fig.3-2 Pull-down style Main Menu Fig.3-3 Toolbar Fig.3-4 Popup Menu

4. Operation We will introduce you how to use M.Wave Professional. 4.1 Quantitative Analysis M.Wave Professional use Standard Curve to test the sample s concentration at a fixed wavelength point. 1. (Fig.4-1) in the Tools Bar shortcut to create a new Quantitative Analysis. Fig.4-1 Quantitative Analysis 2. in the Tools Bar Shortcut to set the parameters of Quantitative Analysis(Fig.4-2). Fig.4-2 Parameters Setting of Quantitative Analysis Choose test method; Input test wavelength in WL Frame; Choose the Concentration Unit ; Choose curve fitting method; Create Standard Curve; Two methods are under your choice to create a new curve. Method 1: Coefficient (1) Click Coefficient option; (2) Click Fit Method to choose Fit Curve method; (3) Input the curve equation s coefficients in corresponding frames; (4) Click OK to finish setting. 3. 4. 5. 6. 7. Fig.4-3 Fit Method Equation s coefficient Method 2: Use Standard Samples to create a new curve

(1) Click Standard Sampling option(fig.4-2); (2) Click Standard Sample Quantity to choose the quantity( 20); (3) Input the sample concentration in corresponding Frame; Fig.4-4 Input Standard Sample Concentration (4) Click OK to finish setting; (5) Pull the Reference into the light path and click the icon, then the system will move the wavelength to the one you just set, it blanks then; (6) Put 1# standard sample into the light path and move the cursor on the fist frame of Abs. The sample Absorbance value will appear in this frame after click in the shortcut toolbar; (7) Measure other standard samples in the same way as step(6) shows. The curve displays automatically when finish(fig.4-5); Fig.4-5 Create Standard Curve and Measure You can modify the curve s coordinates by Clicking the icon.(fig.4-7). You can also save the curve by clicking, and you can reload it by clicking next time. Fig.4-6 Standard Curve Fig.4-7 Rescale the Curve s Coordinates 8. Pull the unknown into the light path (if it is Automatic 8-cell Holder in the instruments, click to set the location of the relative cell first), Then click the icon, click in the shortcut toolbar to test, the test result will display in the data sheet (Fig.4-8).

Fig.4-8 Quantitative Analysis 4.2 Kinetics Analysis We will introduce how to measure a sample s change over a selected period of time. 1. to begin a new test of Kinetics(Fig.4-9); Fig.4-9 Kinetics Analysis 2. Click the button of to set the parameters(fig.4-6); Fig.4-10 Parameters Setting of Kinetics Choose Photometric Display Mode, input the Wavelength, Display Time, Total Time, Intervals, Responsible Mode and Coordinates Values; 4. Click OK to finish and exit setting; 5. Pull the reference into the light path and click icon in the Tool Bar to set zero and full scale. Pull the sample into the light path and click the button of to begin the test. Fig.411 you can click icon in the Tools Bar to cancel at any time. 3.

Fig.4-11 Kinetics Analysis 4.3 Spectrum Scan We ll introduce how to create the spectrum while using Spectrum Scan function. 1. on the Tools Bar to start a new Spectrum scan(fig.4-12); Fig.4-12 Spectrum Scan 2. Scan(Fig.4-11); in the Tool Bar to set the parameters of Wavelength Fig.4-13 Parameters Setting of Spectrum Scan 3. 4. 5. Choose the Photometric Mode, Wavelength Range for Scanning, Intervals, Repeat Times, Response Mode, and Coordinate Range; Click OK to finish and exit setting; Put reference in to the light path, click the icon in the Tool Bar, then the

dialog box of System Baseline displays in the screen (Fig.4-14), click Cancel to interrupt scanning and exit the course; Fig.4-14 Scan Baseline 6. Put the unknown sample in the light path, click the icon in the Tools Bar to start scanning (Fig.4-5), click to interrupt and cancel scanning. Fig.4-15 Spectrum Scan 4.4 Multi-wavelength Analysis We ll introduce how to measure a sample in different wavelengths ( 20). 1. on the Tools Bar, a new form appears on the screen (Fig.4-16); Fig.4-16 Multi-wavelength Analysis 2. on the Tools Bar to set the parameters (Fig.4-17);

Fig.4-17 Parameters Setting of Multi-wavelength Analysis 3. 4. 5. 6. Choose the quantity of the wavelength, you need input the value of each wavelength; Click OK to finish and exit setting; Pull the Reference into the light path, click the icon in the Tools Bar to set zero; Pull the sample into the light path, click to start test. And the test result will be listed in the data sheet(fig.4-18). Fig.4-18 Multi-wavelength Analysis 4.5 DNA/Protein Analysis We will introduce how to operate a DNA/Protein Analysis. 1. on the Tools Bar to begin a DNA/Protein analysis (Fig.4-19); Fig.4-19 DNA/Protein Analysis

2. to set parameters. (Fig.4-20); Fig.4-20 Parameters Setting of DNA/Protein Analysis 3. 4. 5. 6. Choose the measure method first. We provide you 2 methods to measure, and you can also definite the coefficients and wavelength by yourself; Click OK to finish and exit the setting; Pull the reference into the light path, click in the Tools Bar to set zero; Pull the sample in the light path, click the icon to measure. The result will display in the data sheet (Fig.4-27). Fig. 4-21 DNA/Protein Analysis 4.6 Energy Scanning We will introduce how to scan the variability of energy in the range of wavelength. 1. Click Main Menu File New Energy Scan to build energy scanning(fig.4-22); Fig.4-22 Energy Scan

2. on the Tools Bar to set parameter; Fig.4-23 Energy Scan Setup 3. Select Lamp Switch Mode, Wavelength Range, Intervals, Repeat Times and Coordinate Range; 4. 5. Automatic Switch Mode---The instrument can choose D-Lamp or WLamp according to setting; Deuterium Lamp Mode---Using Deuterium Lamp in the whole wavelength; Tungsten Lamp Mode---Using Tungsten Lamp in whole wavelength. Click OK to finish and exit setting; Pull the unknown sample in the light path, click the icon in Tools Bar to start scanning (Fig.4-24), click to interrupt scanning and exit the course; Fig.4-24 Energy Scan 4.7 Modify a Sample 1. Choose the data frame you want to modify or set the curve as current curve; 2. Pull the unknown sample in light path; 3. in Tools Bar to re-measure it, and save the current result to substitute the original one. 4.8 Delete a Test Result 1. Choose the data frame you want to modify or set the curve as current curve; 2. in the Tools Bar to delete the data. 4.9 Name a Sample 1. Select the line you want to rename.

2. Double-click Sample Name, input the sample s name, and press Enter on the keyboard to confirm. 4.10 Curve Processing We will show you the operation of curve. 4.10.1 Switch Display Mode or in the Tools Bar to switch the display mode (Abs. or Transmit). 4.10.2 Zoom Selected Area in Tools Bar, choose the area you want to magnify, then you ll see the magnified one, the Mouse s cursor turns into the form of +. again to cancel the current status. 4.10.3 Modify Scale on Tools Bar to display coordinate as you like. Fig.4-25 Modify Scale 4.10.4 Auto Fitting on Tools Bar to change the scale to appropriate value. 4.10.5 Select as Current Curve Click Main Menu Curve Operate Choose the curve as you like. 4.10.6 Change the Color of Current Spectrum Click Main Menu Curve Operate Choose the color as you like. 4.10.7 Search Peaks in Tools Bar to search the peaks, they labeled automatically on the curve, and the peaks will be listed in the Right Frame. You can set the Peak height by clicking icon. Fig.4-26 Search Peak

Fig.4-27 Search Setup 4.10.8 Mathematics Function s or in the Tools Bar(Fig.4-28, Fig.4-29), a new dialog box will display on the screen respectively. You can add, subtract, multiply or divide two spectrums (Fig.4-28,Fig.4-29). Fig. 4-28 Spectrum Addition Fig. 4-29 Spectrum Derivative Class Select Fig.4-30 Spectrum after Addition Fig.4-31 Spectrum after Derivation 4.10.9 Spectrum Smooth We use this function to wipe off the interferer of noise during the course of scan, so as to make the spectrum look much smoother. Click Main Menu Spectrum Spectrum Smooth

5. Other Functions We will introduce the additional function about M.Wave Professional in this chapter. 5.1 Turn On/Off W Lamp Main Menu Device Turn On/Off W Lamp to open or close W Lamp. 5.2 Turn On/Off D2 Lamp Main Menu Device Turn On/Off D2 Lamp to open or close D2 Lamp. 5.3 Setup Switch Point of Light Source Click Main Menu Device Setup Switch Point of Light Source (Fig.5-1), input the value of wavelength, and then click OK to confirm. Fig. 5-1 Set Switch Point of Light Source 5.4 Set Wavelength on the Tools Bar to set wavelength (Fig.5-2). Input the value of wavelength in the frame of Set Wavelength, click Goto to confirm. Then the instrument will move the wavelength to the point you want, then it blanks automatically. Fig.5-2 Set Wavelength 5.5 Get Dark Current Click Main Menu Device Get Dark Current, the system will reget the dark current and the new data will replace the old one. 5.6 Scan System Baseline Click Main Menu Device System Baseline, the dialogue box will display in the screen (Fig.5-3), you can click Open to choose the storage base line to test, be sure that nothing in the light path, you can click Scan to build system base line. You can click Cancel to interrupt and exit the course. You can choose Save System Base Line option; a dialogue box will display in the screen and click Save to save system base line for your after use. Fig.5-3 Scan System Baseline You needn t to build system base line normally, only when it is a very long time that the instrument wasn t be used or the environment changed much. 5.7 Automatic 8-cells Changer(Accessories) to pull the correspond cell in the light path. 5.8 Cell Calibration(Need install auto 8 cells Changer) Click Main Menu Device Cell Calibration,this operation for matching difference of all the cells selected.

6. File Operation We will introduce how to process the test files in this chapter. Lists of the file s type Quantity Analysis File Kinetics Analysis File Spectrum Scan File Multi-wavelength File DNA/Protein Analysis File Energy Scan File Standard Curve Baseline *.qua *.kin *.wls *.mti *.dna *.ens *.std *.sbl 6.1 Save a Testing in Tools Bar, input the file name and click Save to confirm in the dialog box. 6.2 Open a Testing in Tools Bar, choose the file name you need in the dialog box, then click Open to confirm. 6.3 Print Test Report in Tools Bar, then set Print Parameters in the dialog box, press Print to confirm. 6.4 Export Data to Excel (the Microsoft Excel should be installed) Click Main Menu File Export to Microsoft Excel, Excel will be started up, and data will be stored in it. 6.5 Export Curve as bmp Picture Click Main Menu File Export Bitmap, input the file name and click save to confirm the dialogue box.

Appendix 1 Methods of Quantitative Analysis Single Wavelength Method Abs.=A1 Double Wavelengths Method Abs.= A1-A2 Three Wavelengths Method Abs.=A1-(W1-W2)*(A2-A3)/(W2-W3)-A3 Appendix 2 Methods of DNA/Protein Analysis Method 1 CDNA=(A1-Aref)*f1-(A2-Aref)*f2 CProtein= (A2-Aref)*f3-(A1-Aref)*f4 Ratio=(A1-Aref)/ (A2-Aref) A1=A260nm A2=A280nm Aref=A320nm (Optional) f1=62.9 f2=36.0 f3=1552 f4=757.3 Method 2 CDNA=(A1-Aref)*f1-(A2-Aref)*f2 CProtein= (A2-Aref)*f3-(A1-Aref)*f4 Ratio=(A1-Aref)/ (A2-Aref) A1=A260nm A2=A230nm Aref=A320nm (Optional) f1=49.1 f2=3.48 f3=183 f4=75.8