Manual 517300C SPECTAOSHOTOWETER OPERATING INSTRUCTIONS |S.A80 900uEn 29001 BECKMAN Beckman Instruments, Inc. 2500 Harbor Boulevard * Fullerton, CA 92634-3100 (714)871-4848 * FAX: (714)773-8283 * Telex: 678413 July 1995 ©1995 Beckman Instruments, Inc. Printed in U.S.A.TABLE OF CONTENTS Section One - Introduction 14 12 Section Two - Installation 241 22 23 2A 25 2.6 General Description ... . Principles of Operation . . Installation Instructions Utility Requirements Power Up ... Configuration . Calibration . . . When Not In Use Section Three - General Operating Instructions 3.1 32 33 3.4 3.5 3.6 37 Analysis Windows General Operating Procedure Help Messages ...... The Mouse ....... Permanent Menu Bar Window Menu Bars . Parameter Input... . 3.8 Method Development and Use . . 3.9 Stored Data Section Four - Getting Started 4.1 Power Up 42 RediRead™ Mode a 4d 4.3 RediScan™ Mode o 46 4.4 Fixed Wavelength 48 4.5 Wavelength Scan » 411 4.6 Time Drive .... 4-14 4.7 Recalling Stored Files .................0.0000 000, 417 Section Five - Fixed Wavelength 5.1 Principles of Operation . 5 5.2 Parameter Setup » 52 5.3 Sample Analysis . . 55 5.4 Example Analyses vee 58 5.5 Data Output . . 5-11 5.6 Files ....... 5-13 5.7 ASCII Format 5-13 5.8 Lotus Format ... . 5-14 ContentsSection Six - Wavelength Scan 61 62 63 Principles of Operation. Parameter Setup ......-. : Analysis of Single Samples ... 64 Analysis of Multiple Samples 6-12 65 Data Manipulation . . 6-15 6.6 Function Selection 6-17 6.7 Tabulated Data .... 6-19 6.8 Spectral Addition, Subtraction and Multiplication ae 6-20 6.9 Scatter Correction 6-22 6.10 Net Absorbance Calculations . 6-24 6.11 Example Analyses . 6-26 6.12 Data Output . - 6-30 6.13 Files ....... 6-31 6.14 ASCII Format : : » 6-31 6.15 Lotus Format ......... 20005 -. 6-32 Section Seven - Kinetics/Time 7.1 Principles of Operation .......... 0000 e eee eee eee 12 7.2 Parameter Setup 73° Data Collection 7A Plotted Data .. 7.5 Rate Calculation 7.6 Raw Data ...... qd 78 19 7.10 ASCII Format . Example Analyses Data Output Files 7.11 Lotus Format ... Section Eight - File Utilities 8.1 8.2 83 89 8.10 Receive Command File Utility Window .. Directories ....... Rename Command . Copy Command ... Move Command .. . Delete Command . . Disc Status Command Format Command . Convert Command . 8.11 Transmit Command . . ContentsSection Nine - Data /O 9.1 Communications Configuration . . 9.2 Diagnostics . © 94 9.3 Output Mode .. «+ 9-8 9.4 Remote Control Mode ... vee OD 95 Remote Control Commands . - 911 9.6 Remote Examples 9-17 Section Ten - Preventative Maintenance 10.1 General Information 10-1 10.2 Status Window 10-2 Section Eleven - Troubleshooting 11.1 Power Up Diagnostics 11.2 Operational Failures . 11.3 Operational Messages Section Twelve - Corrective Maintenance 12.1 Fuse Replacement 12-1 12.2 UV Source Replacement . 123 12.3. Visible Source Replacement . . 12-7 Section Thirteen - Technical Specifications 13.1 Performance Specifications 13-1 13.2 Physical and Environmental Specifications . 13-2 13.3 Storage and Transport 13.4 Sample Compartment Configuration . . 13-3 Section Fourteen - Parts, Supplies and Accessories ............ 14-1 Section Fifteen - Beckman Sales and Service Offices .......... 15-1 Section Sixteen - Warranty ........ 20.20... ccc cece eee ee 16-1 Index oe ee ene teen eee 17-1 Contents iitSECTION ONE INTRODUCTION 1.1 General Description The DU® Series 600 Spectrophotometer is a microprocessor controlled spectrophotometer intended for use in quantitative and qualitative biological research and industrial procedures that require spectrophotometric measurements in the UV-visible region of the electromagnetic spectrum. If the instrument is used in a manner other than as described, the safety and performance of the instrument can be impaired. The DU Series 600 Spectrophotometer operates in the wavelength range of 190 to 1100 nm. Models of the instrument are available with cither a monochrome or a color video display. Data storage on a 3% inch diskette is optional. Various optional accessories are available, to configure the instrument for specific application needs. The instrument features a graphic video display, which provides operational information using windowing techniques, and a "mouse" for operator control. The mouse js used to position an arrow on the window. When the arrow points to the desired position, the left button on the mouse is pressed to initiate the desired action. In these instructions, the positioning of the arrow and pressing the left mouse button is called "clicking on". ‘The instrument has two. rapid reading modes: RediRead” for taking readings at a fixed wavelength and RediScan” for making a wavelength scan, The instrument has three standard Routine Measurement modes. They include: Fixed Wavelength - Takes absorbance or transmittance readings at up to 12 wavelengths. Readings at each wavelength can be multiplied by a factor. Wavelength Scan - Performs wavelength scans in absorbance or transmittance. Data are automatically stored for manipulations includ- ing Trace, zoom, overlay and tabulate. Calculations include peak pick, Introduction itvalley pick, point pick, first to fourth derivative, log of absorbance, scatter correction, spectral addition, subtraction and multiplication, and net absorbance. Repetitive scanning is also performed in this mode. Kinetics/Time - Calculates the rate of an absorbance versus time reaction with a choice of blank subtraction and graphic display of the data for multiple samples. Data are automatically stored for * manipulations including Trace, zoom, overlay and tabulate. Other Application modes are also available, which include: 12 Protein Analysis - Calculates the protein concentration using the Bradford, Lowry, Biuret, direct UV method, colloidal gold, and bicinchoninate (BCA). Prepares a standard curve using up to 30 stan- dards. The user can choose to add, delete or rerun individual standards based upon a statistical analysis of the standard curve. Nucleic Acid - Determines protein impurity in nucleic acid samples based upon the ratio of readings at two wavelengths with a choice of background correction. Protein and nucleic acid concentrations can also be calculated using the Warburg and Christian‘ coefficients. Fraction Read/Plot - Collects, plots and tabulates data from a set of related fractions. Readings can be corrected for dilution. After data collection, individual fractions can be added, deleted or rerun. Data are plotted versus either fraction number or volume. Other data obtained for the fractions can be input and plotted with the absorbance data. Single component quantitative analysis - Calculates a standard curve from up to 30 standards using either linear or nonlinear least-squares regression. Performs statistical analysis on the standard data. Allows the user to add, delete or rerun individual standards to optimize the calibration. Calculates the concentration of samples from the calibration data. Enzyme Mechanism - Guides the operator through the necessary steps to calculate Km and Vmax. Displays the following plot types: Michaelis-Menten, Lineweaver-Burk, Eadie-Hofstee, and Hanes-Woolf. Calculates the Hill constant from the Hilf Plot. Graphs inhibitor plots to determine Ki. ‘Warburg, O. and Christian, W., Biochem. Z:310, p. 384f (1942). Introductionsamples. Multicomponent analysis - Uses Full Spectrum Quantitation (FSQ™) to calculate the concentration of up to ten components in a mixture from up to 32 standards which are mixtures. Performs a statistical analysis of the standard data to determine the accuracy of the calibration. Allows individuat standards to be added, deleted or rerun to optimize the calibration. Calculates the concentration of samples from the calibration data. Gel Scan - Collects and plots absorbance data as a function of distance for a sample prepared by electrophoresis. Calculates peak and valley locations, which are used for subsequent area calculation or molecular weight determination. Performance Validation - Provides a simple procedure to verify the performance of an instrument. Tests which are performed include: wavelength accuracy and repeatability, resolution, baseline flatness, noise and stability. One model of the instrument also contains a program mode, which allows the user to customize applications by writing programs which blank, collect and store data at the desired wavelengths, calculate results from the data, prompt the operator, format and label the output and control sampling accessories. The instrument is provided with a paraliel output for a Dot Matrix Printer and a bidirectional RS-232 communications port that can be configured for either communications or an X-Y Plotter. The communications port can be used for data transfer to and from the instrument and for remote control of the instrument by an external computer. A full line of modular sampling accessories is available for the instrument. Included are temperature controlled cell holders, automatic multi-position cell holders, a batch sampler, sipper samplers, and long pathlength cell holders. The automated sampling accessories (sipper samplers, batch sampler, and auto samplers) are designed to operate in the modes described previously to automate analyses. Microsampling capabilities include the use of the 50 wl. Microcell, the 100 pL Multi-Microcell, and the 5 pL Ultra- Microcell. Introduction 1312 Principles of Operation Optical Principle The optical diagram of the DU Series 600 Spectrophotometer is shown in Figure 1-1. @ SEUTERIUM source ruussren cca COMBIKER, SOURCE aH Prove MWRFOR TRANSFER, ‘TRANSFER MIRROR } wianon ENTRANCE su FILTER ox & si MIRROR ‘SAMPLE COMPARTMENT GRATING Figure 1-1, DU Series 600 Spectrophotometer Optical Diagram The DU Series 600 Spectrophotometer is a single beam spectrophotometer. Light from both sources enters the monochromator, where it is dispersed by the concave holographic grating. Monochromatic light exits the monochromator and illuminates the sample. The amount of light that passes through the sample is measured by the single photodiode detector. The focal point of the beam in the sample compartment is on the right-hand side. This location permits the maximum amount of transmitted light to reach the detector from scattering samples. All sampling accessories position the sample at the focal point for best performance with | microsamples and gels. 14 IntroductionBlanking Method A blank is always required before data collection; any reading taken without a blank is invalid. A blank reading is taken when <> (located in the permanent menu bar on the bottom of the window) is clicked on. NOTICE In the RediRead” Mode the blank command is . In the RediScan™ Mode, the blank command is . When the instrument blanks, the following steps are performed: 1. The monochromator is moved to the proper wavelength. This is the specified wavelength for a single wavelength reading. 2. The proper detector gain value is selected automatically. This minimizes the noise level and maximizes photometric accuracy. 3. Dark current is measured and corrected. This compensation assures accurate readings at high absorbance. 4. In the Wavelength Scan mode, only, a background scan is made. The blank (or reference) is automatically scanned over the same range at the same speed that the samples will be scanned, so that the background correction is optimal. This calibration assures repeatable readings every time the instrument is used. In all modes, a blank solution should be in the sample compartment during the blank. It is suggested that the solvent used to prepare the samples be used for the blank. However, air (no sample) may be used. A new blank reading should be taken each time the solvent is changed. NOTICE Plastic cuvettes, glass (Pyrex) cuvettes, and some solvents have significant absorption in the UV region. Verify that they transmit UV light by scanning them versus air before using them in the UV region. To re-zero the instrument at any time between samples, insert the same blank solution and click on <>. Introduction 1-5The instrument stores the blank and uses it until cither the sources are turned off or another blank reading is taken. For best results, the instrument should be blanked frequently, allowing the blank reading to be taken shortly before the sample measurement is taken. A new blank should be read if the instrument has not been used for an hour. Scanning The background scan, made as part of the Blank procedure, is stored in the instrument and can be reused for an unlimited number of sample scans as Jong as the range and scan speed remain the same; unless the user blanks, the sources are turned off or the instrument is turned off. (The range can be decreased as long as the scan speed remains constant and no new back- ground is required.) When a new background is required, it is indicated on the display. A new background scan should be made every time a solvent is changed, because the background spectrum will likely be different. A new background scan should also be made if no scan has been made for over an hour. To rescan the background, click on <>, while in the Wavelength Scan mode. The selected scanning speed determines the distance between each data point that is collected as the instrument scans through the chosen region. At 1200 nm/min, a data point is collected every nanometer. At 600 nm/min, a data point is collected every half nanometer. As the sample data are collected the background is subtracted and the difference in absorbance (or ratio in transmittance) is plotted on the display. Read Average Time The noise level of the instrument, and therefore the uncertainty of a sample reading, is decreased by taking a number of readings and averaging them. The instrument takes a reading every 0.05 second. It takes a series of these readings over a user-selected time and averages them to obtain the blank and sample readings. For example, with a read average time of 0.5 seconds, ten readings are taken and averaged. The operator can specify a read average time from 0.05 to 99.9 seconds in all modes except Wavelength Scan and Multicomponent Analysis. 16 IntroductionRead averaging is not used in the RediScan and Wavelength Scan modes, Background and sample scans are collected without averaging. Smoothing is used to improve the appearance of the data. | | Smoothing The displayed wavelength scan can be smoothed using a selectable smoothing function. The calculation, using the Savitzky and Golay* coefficients (as modified for end points by Peter A. Gorry’), is done for every data point in the scan, using the data points before and after the point of interest. The user selects the total number of data points used for the calculation, from 7 to 25. Use of too few points does not rid the scan | of noise. Use of too many points can cause real peaks to be combined. *savitzky, A., and Golay, M., Anal Chem, 36, 1964, p1627f. SGorry, Peter A., Anal Chem, 62, 1990, p570f. IntroductionSECTION TWO INSTALLATION 2.1 Installation Instructions The DU Series 600 Spectrophotometer is user installable. As an option, it can be installed by a qualified Beckman Field Service Engineer. The instructions for user installation are provided in Manual 517315. Location ‘The DU Series 600 Spectrophotometer is designed to sit on a lab bench or table, which is level and flat and is capable of supporting its weight and the weight of all accessories. ‘The instrument is designed to operate in a clean laboratory environment, free from dust, fumes, excessive moisture, and corrosive chemicals. It should not be exposed to drafts from heating and cooling vents, heating elements, open windows or doors, Lab areas that receive direct sunlight should also be avoided. An ambient room temperature of 15 - 40°C (59 - 104°F) should be maintained. Relative humidity should be 85% or less. Instrument performance can be affected by strong electromagnetic fields that can exist in the proximity of large electric motors, centrifuges, diathermy machines and microwave sources. The batch sampler must be placed adjacent to the right-hand side of the instrument. The other accessories can be placed in a convenient place, near the instrument, and within reach of the cables. Additional space is required for air circulation around the instrument and accessories for proper performance. Do not block these air spaces. Installation ad | i | | | | | | |2.2 Utility Requirements Each of the following needs its own electrical outlet: the spectrophotome- ter, display, dot matrix printer, X-Y Plotter, batch sampler, and Peltier temperature controller. The electrical requirements of each are summarized in Table 1 for 100/120V and Table 2 for 220/240V systems. The same circuit should be used for the instrument and all accessories. A dedicated circuit is preferred. Do not use a circuit which is also used by equipment that operates intermittently and creates wide fluctuations in power demand, such as refrigerators, water baths and centrifuges. For 100/120V operation. The power line should provide three-wire single phase power. To provide multiple outlets, grounding type power strips may be used. Extension cords or multiple outlet adapters should not be used. For 220/240V operation. The power line should provide three-wire single phase power. To provide multiple outlets, grounding type power strips may be used. Extension cords or multiple outlet adapters should not be used. 22 InstallationFrequency —_—_-Voltage Current (Hz) (VAC) (Amps) Spectrophotometer 50/60 100/120V+10% 18 Display 50/60 110V+10% 08 Dot Matrix Printer 50/60 120V+10% 0.6 X-Y Plotter 48 - 66 120V -10%,+5% 02 Batch Sampler 50/60 117V+10% 0.8 Peltier Temperature Controller 50/60 120V+10% 1.0 Table 1. Electrical Requirements for 100/120V Systems Frequency Voltage Current (iz) (VAC) (Amps) Spectrophotometer 50/60 220/240V+10% 0.9 Display 50/60 220V+10% 04 Dot Matrix Printer 50/60 220/240V+10% 0.3, X-Y Plotter 48 - 66 240V -10%,+5% 01 Batch Sampler 50/60 234V+10% 04 Peltier Temperature Controller 50/60 220/240V+ 10% 05 Table 2. Electrical Requirements for 220/240V Systems Installation 232.3 Power Up The DU Series 600 Spectrophotometer is powered up using the following steps. i. Verify that the voltage on the rating plates on the instrument, display and printer are the same as the power source. AS caution The voltage indicated on the display, the instrument and the printer must be the same as the power source. If the incorrect voltage is indicated, do not plug in the instrument and contact the local Beckman service office. The instrument, display and printer must be plugged into grounded power outlets. NOTICE If a power cord is not supplied, contact the local Beckman service office. 2. Plug the power cords on the instrument, the display and the printer into grounded, three-prong outlets that are on the same power line. 3. Turn on the display. The indicator light on the front of the display should illuminate. 4, Turn on the instrument. The power switch is located on the right-hand side of the back of the instrument. The instrument fan should turn on. If the fan does not turn on, turn off the instrument and check the fuses. Directions for fuse replacement are provided in the Corrective Maintenance section of this manual. 5. The display should illuminate, a message window with "Executing Power Up Diagnostics" should be displayed, and an arrow should appear on the display. The arrow should move when the mouse is moved. If the display does not illuminate: a. Verify that the indicator light on the display is illuminated, showing that there is power to the display. 2d Installationb. Verify that the brightness control on the display is adjusted properly. ¢. Verify that the cable on the display is connected to the "DISPLAY" port on the back of the instrument. If the mouse does not move the arrow, verify that the cable attached to the mouse is connected to the "MOUSE" port on the back of the instrument. NOTICE If the recommended action does not correct the problem, power down the instrument, then power it up again. If the problem persists, contact the local Beckman service office. 6. Turn on the printer using the switch on the left-hand side of the printer. Verify that the "ON LINE" light is illuminated (or blinking). 7, Verify that the printer paper is loaded and that the top of the first page is aligned properly. NOTICE The instructions for loading of the paper and the replacement of the ribbon are located in Manual 514521. Installation: 252.4 Configuration After the DU Series 600 Spectrophotometer has been installed and powered up, the Configuration mode is used to select parameters which setup the instrument, the output devices, and the sampling accessories. It is also used to assign the passwords for method protection. Configuration parameters are generally selected when the instrument is installed and are not changed during operation of the instrument. The DU Series 600 Spectrophotometer user interface operates on the principle of windows. The "mouse" is used to position an arrow on the window. When the arrow points to the desired position, the left button on the mouse is pressed to initiate the desired action. In these instructions, the positioning of the arrow and pressing the left mouse button is called “clicking on". If the Power Up Diagnostic window is displayed, click on to remove the window and display the Main window. The Configuration window, Figure 2-1, is used to select one of seven windows that are used for different configurations. ‘It is displayed when "CONFIGURATION" is clicked on from the Main window. eAEUTEIETAD Userinterface Print/Plot Clock accessory Communication Protection BECKHAN DU-600 CORFIGURATION Figure 2-1. Configuration Window 2.6 InstallationNOTICE Jastructions to configure the instrument and Printer/Plotter, only, are provided in this section. Instructions for the X-Y Plotter are provided in Manual 514523. Instructions for the sampling devices are provided in Manual 517314. User Interface Configuration The User Interface Configuration window, Figure 2-2, is displayed when is clicked on from the Configuration window. eer Intertare Color Palette: Blues Screen saver delay: 15 min Screen keyboard image: (Typewriter) Plot grid enable: [Yes] Program to run after power up: Automatically mark data file for save: [No ] OK Figure 2-2. User Interface Configuration Window Set the following: 1, Color palette - This determines the set of colors used on the display for a color monitor, or if a monochrome monitor is used. Click on the current selection to display a window with the palette options. Click on the box before the palette of choice, then [OK] to remove the window. 2. Screen saver delay (minutes) « The instrument dims the display whenever the instrument is not used for the selected amount of time and both sources are turned off. Click on and input the desired amount of time. 3. Screen keyboard image - This determines the order of keys on the alphanumeric keypad. Click on to toggle between [Typewriter] and [Alphabetic]. 4. Plot grid enable - This determines whether a grid is displayed on graphs. Click on to toggle between [Yes] and [No] to enable or disable the grid, respectively. Tstallation 2The input to this selection determines whether the grid is plotted on printouts generated on the Dot Matrix Printer: For printouts on the X- Y Plotter, however, the grid is enabled or disabled on the Printer and Plotter Configuration window, 35. Program to run after power up - This allows a user-written program to _ be executed automatically after power up is completed. The program could turn on the sources, have a delay to allow the sources to warm up, then execute the Performance Validation tests. (The Program mode, which is used to write the program, is not included on all models.) To select a program, click on the displayed program name to display the Program directory. Click on one of the program names to select it. 6. Automatically mark data file for save - This affects the Save Clear and Quit windows, only. If this is enabled, the box is automatically darkened for each data file when the window is displayed. If this is disabled, the box is darkened when it is clicked on or if the file name is input. When the desired parameters are selected, click on [OK] to store the entries and remove the User Interface Configuration window from the , display. Clock Configuration The Clock Configuration window, Figure 2-3, is displayed when is clicked on from the Configuration window. CLOCK SET: Set time: 13:30:22 Set date: 10/25/91 Displayed time format: [24 hours] Date format: [mmddyy] WAKE UP: Yake up enabled: [No] Wake up time: 07:00:08 Program to pun at wake up: Figure 2-3. Clock Configuration Window i | | 28 InstallationCLOCK SET Input the following information to set the date and time, displayed in the permanent menu bar. 1. Set time - Input the current time, using the 24 hour clock. The clock starts when [OK] is clicked on to remove the window. Set date - Input the current date. Time format - Select format for the time: [12 hours] or [24 hours}. If a 12 hour clock is selected, am and pm indications are not given. Date format - Select format for the date: [mmddyy], [ddmmyy], or [yymmdd]. WAKE UP This allows the instrument to turn on the sources at an operator determined time, so that the instrument and the sources can be warmed up and ready to use. The instrument should be left with the Main window displayed and the sources turned off in preparation for Wake Up to execute. 5. Wake up enabled - Toggle between [Yes] and [No] to enable or disable the wake up mode. Wake up time - Input the wake up time using the 24-hour clock. Program to run at wake up - This allows a user-written program to be executed automatically after the sources are turned on. The program could have a delay to allow the sources to warm up, followed by the Performance Validation mode or some other diagnostic tests. (The Program mode, which is used to write the program, is not included on all models.) To select a program, click on the displayed program name to display the Program directory. Click on one of the program names to select it, When the desired parameters are selected, click on [OK] to store the entries, start the clock and remove the Clock Configuration window from the display. Installation 29Protection Configuration The Protection Configuration window is used to identify each user and assign a password, The password prevents methods from being modified by another user. The master access code is required before passwords can be assigned or changed. This prevents unauthorized change of password. To input or change passwords: 1. Click on from the Configuration window. The Protection Configuration window, Figure 2-4, is displayed. CSR ee sea Naster access code: XXXXXXK (ok) Figure 2-4. Protection Configuration Window 2. Click on the X’s that are displayed for the master access code, input the code, then click on [OK]. The Edit User Names and Passwords window, Figure 2-5, is displayed. NOTICE The master access code is assigned by Beckman Instruments and cannot be changed. PELE SET ao) User Nane Password « Frank Aralis Fa2 . Tsafpina Meir Barry Master . Mike Simonian . Linda Wheeler . Valerie Kohler Figure 2-5. Edit User Names and Passwords Window 2410 Installation | : i3. Click on a position in the table in the "User Name" column and input a user name. Then click on the corresponding position in the "Password" column and input a password. Repeat for each user. 4. To edit any of the entries, click on it and input the desired information. 5. When all of the information has been input correctly, click on [OK] to store the information and remove the Edit User Names and Passwords window from the display. Then click on [OK] to remove the Protection Configuration window from the display. Printer Configuration The Printer and Plotter Configuration window, Figure 2-6, is displayed when is clicked on from the Configuration window. Printer and Platie onfigurst Printer Drivers Epson FX-858 Titles BECKMAN DU-690 Subtitle: Send graphs to: [Printer] Send screen to: [Bit Image File) XY PLOTTER: Grid: (Off) Number of pens: 8 Figure 2-6. Printer and Plotter Configuration Window Select the following parameters: 1. Printer Driver - Selects the printer type being used, If a HP Desk Jet is selected, all text screens with 80 characters or less are printed portrait, and all graphs and all text screens with more than 80 characters are printed landscape, and, if any line has more than 100 characters, the print is condensed. To use the Cannon BJ-200, select the Epson LQ-510 and make sure that the DIP switches are set for the Epson LQ-510 mode. 2. Title - Input up to 60 alphanumeric characters which are printed at the top of each printout made with the command. This can be used to identify the institution or department where the data were generated. Installation 2A3. Subtitle - Input up to 60 alphanumeric characters which are printed on the second line of each printout made with the command. This can be used to provide an analysis description or 1o identify the laboratory where the data were generated. 4, Send graph to - If both the printer and plotter are installed, select from [Printer] and [XY Plotter]. This option determines where a printout, which includes a graph, is printed. 5. Send screen to ~ This determines where a screen copy is sent when [PrtSern] is clicked on. Select from either [Printer] or {Bit Image File]. If [Bit Image File] is selected, the information on the screen is made into a TIFF file. The TIFF file is named by the user and the file is placed in the ASCII directory. 6. Grid - This parameter is used for the X-Y Plotter, only. Refer to Manual 514523 for more information. 7. Number of pens - This parameter is used for the X-Y Plotter, only. Refer to Manual 514523 for more information. When the desired parameters are input, click on [OK] to store the entries and remove the Printer and Plotter Configuration window from the display, then to return to the Main window. 212 Installation2.5 Calibration There are three calibrations that are performed by the DU Series 600 Spectrophotometer: absorbance (or transmittance), wavelength and scan gain. Absorbance (or transmittance) - The absorbance reading is calibrated each time a blank reading is taken. Blanking is discussed in section 1.2. Wavelength - The wavelength is calibrated at the factory prior to shipment. At any time after installation, the wavelength can be recalibrated using the following instructions: 1. Verify that both sources are turned on. If either is off, turn it on by clicking on the appropriate command in the permanent menu bar. 2. With the Main window displayed, click on "DIAGNOSTICS" to display the Diagnostics window. 3. Click on . The instrument finds the deuterium emission line at 656.1 nm, recalibrates the wavelength and stores the information. (The instrument also checks the scan gain values and adjusts them, if necessary. Scan gain is discussed below.) 4. When the wavelength calibration is complete, click on to remove the Diagnostics window and display the Main window. Under normal operating conditions, it should not be necessary to recalibrate the wavelength on a routine basis. However, it may be desirable to recalibrate the wavelength after moving the instrument. Scan Gain The scan gain is set at the factory prior to shipment. The gain is checked and adjusted each time that the wavelength is calibrated, as described above. When a wavelength scan is performed using a blank that has significant absorbance in the scanning range, it may be desirable to change the scan gain settings in the instrument to compensate for the absorbance of the blank. This will increase the dynamic range of the instrument when scanning using the absorbing blank. To change the scan gain: 1. With the Main window displayed, click on "DIAGNOSTICS" to display the Diagnostics window. Installation 213 | i2. Place a cuvette of the blank in the cell holder. 3. Click on . The instrument adjusts the scan gain values 1o compensate for the absorbance of the blank. 4, When the scan gain has been set, click on to remove the Diagnostics window and display the Main window. NOTICE The instrument will continue to use the new gain values until the gain is adjusted using either or . It may be desirable to repeat the above procedure with nothing in the sample compartment after changing the gain for a significantly absorbing blank. 2.6 When Not in Use When the instrument is not in use for more than two hours, turn off both sources. The power to the instrument can, but does not need to, remain turned on. The “Screen Saver Delay". set in the User Interface Configuration Window, dims the display after the selected time. Any movement of the mouse will re-ifluminate the display. Before using the instrument again, turn on either or both sources,” depending upon the application, and allow them to warm up for at least 30 minutes before taking readings. 214 InstallationSECTION THREE GENERAL OPERATING INFORMATION The DU Series 600 Spectrophotometer user interface operates on the principle of windows. A "mouse" is used to move an arrow around a window. Action is initiated when a mouse button is clicked on. The instrument contains several analysis modes, which are selected from the Main window by using the mouse to position an arrow on the mode name and clicking on the left mouse button. When the mode is selected, an analysis window for the mode is displayed. Analysis parameters can be input in the analysis window or can be recalled from a stored method. The analysis modes have an associated Method window, which is used to recall stored methods and to input the parameters to develop new methods. The methods, sample data, calculated results, and standard data can be stored, in a designated file. These files are located in the main instrument or in an optional external disk. Operating Information BA 1 | | | :3.1 Analysis Windows Analysis windows are displayed during normal operation. These windows are used to select parameters, collect data and calculate results. A typical analysis window is shown in Figure 3-1. S>] Results #11 Fo] Read averag: Method neae: ar 1 Cabs} Sampling eeleg y 380.7 a ATE Factor 56,00 Factor 230.0 Abs Result Abs Abs. 9.2790 15. . 6.3152 53 93647 26: "037 ! 63784 8.5747 1224 : 10832 1242 18; 76 9.6413 as86a 118447 114303 O.9504 5312246 86.8482 1.3883 Rediscay Devices 486.0 nn TINE UATE TeMF CELL |. Read PrtScen @.9Bi3 ADs 1:36 12/38/91 N/A BLANK TUIS GFF WRTCH OFF [UV OFF Figure 3-1. Typical Analysis Window The manual uses the following nomenclature when referring to analysis windows: 1- 2. 3. 3-2 Window The entire display, or any portion of the display that is enclosed with a box. More than one window can be displayed at a time. Window Name All analysis windows have a name on the top line of the window. An example of each analysis window is provided in this manual. A listing of these windows is provided at the end of the Table of Contents. Help Messages "HELP" is displayed on the top line of the window, to the right of the window name. Click on "HELP" to display Help windows. Operating Information4- Permanent Menu Bar A list of commands is always displayed at the bottom of the display. A permanent menu bar command is referred to in these instructions with | the use of double angle brackets, ic. <>. Status informa- | tion is provided on the right-hand side of the permanent menu bar. / 3 - Window Menu Bar A list of commands is located at the top of each window, directly below the window name. The specific commands change with each window. A window menu bar command is referred to in these instructions with the use of single angle brackets, ic. . Commands that can be used are identified by color. 6 - File Names The files where the method, standard data, and/or results data are stored are listed directly under the window menu bar. | 7 - Parameters | The analysis parameters are listed near the top of a window. Parameters are referred to in these instructions with the use of quotes, ie. "Wavelength". The values for parameters that can be changed are displayed in a different color than the parameter name. 8 - Data Column Labels Some column labels can be changed, such as the units for calculated results, Labels that can be changed are displayed in the same color as parameters that can be changed. 9 - Scrolling Arrows Click on the arrows to display data that is not displayed because of insufficient room. The window shown in Figure 3-1 has both up and down arrows and right and left arrows. Many windows have only up and down arrows. i i i i | | | | | Operating Information 333.2. General Operating Procedure When the DU Series 600 Spectrophotometer is powered up, it performs a series of diagnostic tests. When these tests are completed satisfactorily, the Power Up Diagnostics window, Figure 3-2, is displayed. Computer and Hardware Diagnostics: Passed PROM RAM Controller Ran Video Controller Video RAN Video Palette RS232 Ports i and 2 EE PROM Passed Spectrophotoneter and Sustens Diagnostics: PROM Option Passed Software Option Ran Option Keyboard Processor Detector ain Welorg aan> Ligne. te | i Lamp sete Waveleni ti Brive Systea Clo Passed Figure 3-2. Power Up Window If all power up tests pass, remove the Power Up Diagnostics window from the display by using the mouse to move the arrow to and clicking on the left mouse button. The Main window, Figure 3-3, is displayed. The Main window is used to select an operating mode. NOTICE If any power up test fails, refer to the Troubleshooting section for instructions. Programmability is an option and may not be installed on all instruments. | | a4 Operating InformationReutene A aunars DURUELENGTH y | | WAVELENGTH SCAN RACTION READ/PLOT PELE UTILITIES IC CID CONFLGURETION SIAGNOSTICS KINETICS? TINE WELE ‘BLANK TWIS_OFFY Regiscan DEVICES “0 on TIRE DATE ——TERP CELT HATCH OFF CUV OFF 3 RediRead PrtScrn 19 abs e123? 12/30/91 NAL Figure 33. Main Window / ‘The following are general operating instructions. Specific instructions for each analysis mode are provided in the respective section in this manual, 1. To select an analysis mode from the Main window, use the mouse to move the arrow to the desired mode. Press the left mouse button to click on the analysis mode. An analysis window, such as shown in | Figure 3-1, is displayed. i 2. Analysis parameters are listed on the window. The mouse is used to input different values for the parameters. More information on parameter input is given in section 3.7. As an alternative, a stored method can be recalled or a new method can be developed. Either of these is done by clicking on , Jocated on the menu bar at the top of the window to display the Method window. A typical Method window is shown in Figure 3-16. 3. When the desired parameters are displayed, place a cuvette of blank solution in the sample compartment and click on <>. (<> is located in the permanent menu bar.) Operating Information 354. Place the first sample in the sample compartment and click on . ( is located on the memn bar at the top of the window.) As an alternative, the right mouse button can be used to take a sample reading, with the cursor in any position. 5. To print the data, click on . All information in the window is printed, even if only partial information is displayed, because of insufficient room. If the window contains graphic data, it is printed on the device selected in the Printer and Plotter Configuration window. If the window contains no graphic data, it is printed on the printer, if installed and operational. The permanent menu bar is not printed. To stop printing while the DU-600 is transferring the information to the printer, the Stop Printing window is placed at the top of the display. To stop printing, click on [QUIT]. Reset the paper to the top of a new page before starting another printout. | To stop printing after the Stop Print window is removed, click on <> to dispiay the Device Control window. Click on i [STOP PRINTING], then to remove the Device Control : window from the display. Reset the paper to the top of a new page I before starting another printout. | 6. To clear all data from the window, with the option of storing the data, | click on . If the data are stored, no additional data can i be placed in the data file. | 7, When the analysis is complete, click on to return to the Main window. ( is located on the menu bar at the top of the window.) An opportunity is given to store the method and/or sample i data. 36 Operating Information33 Help Messages Help windows are displayed by clicking on "HELP", displayed to the right of the window name. Typical Help windows are displayed in Figures 3-4 and 3-5. If a Help window similar to Figure 3-4 is displayed, click on the desired selection to display a Help window similar to Figure 3-5, Click on to remove the Help windows from the display. Help: Main Wing Click on a topic. General Permanent Custom App Remote Con Nethods are stored autonatically when you exit the Method window. Data files cen be stored using . Once stored, neu deta cannot be added to the files (except in Fraction Read}. All files can be stored when you teeve the operating node; . Unen the Save window is displayed: . Click on the box next to the file type to save. . Enter a different file nane, if desired. Default file names nust be changed to save a file. . Click on OK to store the changes. CANCEL returns to previous vindow without saving. Figure 3-5. Help Window Operating Information 373.4 The Mouse The mouse, Figure 3-6, is used to move an arrow to desired locations on the DU Series 600 Spectrophotometer display. When the arrow is in a desired location, click on the left mouse button to initiate action. The items that can be clicked on include window menu bar commands, file names, parameters, sample data, and permanent menu bar commands. Figure 3-6. Mouse : The left mouse button is used the most frequently to click on a command or parameter. However, the mouse buttons have other uses: 1. To take a reading - The right mouse button can be used to initiate a reading, with the cursor in any position, as an alternate to in the window menu bar. 2. To input lower case letters - When the alphanumeric keypad is displayed for character input, click on the right mouse button, rather than the left button, to input lower case letters. 3. To use Trace - Trace is a feature that is used to find the ordinate and abscissa values from graphic data. Trace is a window menu bar command () that is clicked on in the normal manner. After is clicked on, the mouse is used to move the arrow to the! desired location on the graph and the center mouse button is clicked on to position a line on the graph. Then the right and left mouse buttons can be used to move the line to the right and ieft, respectively. 38 Operating InformationThe Hour Glass Most of the time the arrow appears on the window, However, when an action is initiated that cannot be completed quickly, the arrow is changed into an hour glass. The hour glass can be moved across the window in the same manner as the arrow, and the mouse buttons can be clicked on, but no action occurs until the arrow returns. Typical actions that cause the hour glass to be displayed include printing a window and performing complex data calculations. The Diskette Symbol When the optional disk drive is accessed, the arrow is changed into a diskette symbol. The diskette symbo{ cannot be moved and no action can be taken until disk access is completed. Operating Information 393.5 Permanent Menu Bar The permanent menu bar, Figure 3-7, is always displayed at the bottom of the display. Commands that can be used at any time during the operation of the instrument are listed on the left-hand side of the permanent menu bar. Current nanometer position and reading are displayed in the middle. Status information is displayed on the right-hand side. If a source burns out during operation, an error message is displayed under the commands on the left-hand side. LANK 1UES OFF) BLaw Devites MATCH OFF [UU OFF I TEMP CELL Potscen 9. 1 on DATE. abe 01:37 12/30/91 Hea Figure 3-7. Permanent Menu Bar ‘The permanent menu bar commands are: BLANK Take a reading at the analytical and background wavelength(s) and set the value 0.000 absorbance and 100%T. In the Wavelength Scan mode, only, make a background scan, MATCH OFF/ON If an Auto Cell Holder is used for the analysis, zero readings can be taken on each of the cuvettes. When Match is enabled, the zero readings are subtracted from all subsequent readings for the appropriate cell, Operational information is provided in Manual 517314. VIS OFF/ON Turn the visible source on or off. UV OFF/WAIT/ON Turn the UV source on or off. The UV source requires about 30 seconds to light after being turned on. During this time < > is displayed. NOTICE Do not blank the instrument or take sample readings while <> is displayed, even in the visible region. When the UV source lights, readings at all wavelengths are affected. RediScan Enter the RediScan mode. This is described in section 4.3. 310 Operating Informatién| RediRead Enter the RediRead mode. This is described in section 4.2. | DEVICES Display the Device Control window, Figure 3-8. The window is used to stop sending information to the printer or plotter after printing has begun; to position the transport at a ceil position for the Auto Cell Holder, a millimeter position, or the home position to align; to control the temperature controller; and to move the aspirator arm on the batch sampler. When the desired action has been taken, click on to remove the window from the display. device is automatically controlled as part of an analysis. STOP PRINTING. STOP PLOTTING TRANSPORT? single] NAIA) 22) el a) 2] rome] Position: 28.790 mm | 1 | The information input in the Device Control window is overridden if the | | | | | ! i | TEMPERATURE CONTROLLER: Eneble Temperature Controller (No J] I Temperature setting: 30.6 C BATCH SAMPLER: RAISE ARH [LOWER aRW ADVANCE] ARM TO WASH, ARM TO SAMPLE Figure 3-8. Device Control Window PrtSern Print an exact copy of the entire display on the Dot Matrix Printer. Information that is not displayed because of insufficient room will not be printed. The X-Y Plotter cannot be used for this printout. NOTICE There is also a command in the menu bar at the top of most windows, which is used to print the window in which it appears and also prints the continuation of data that are not displayed. i Operating Information sd |The status information includes: Nanometer Position The current nanometer position, Absorbance/Transmittance Reading The current reading, in either absorbance or transmittance, determined by the reading mode in the selected analysis mode. This field is blank until the instrument is blanked. TIME The current time. The time is set in the configuration mode, using the Clock Configuration window. DATE The current date. The date is set in the configuration mode, using the Clock Configuration window. TEMP ‘The current temperature, if the Temperature Controller is being used. When the Temperature Controller is controlling temperature, "on" is displayed under the temperature. If the Temperature Controller is not installed or is not turned on, "N/A" is displayed. The Temperature Controller is turned on using the Device Control window, Figure 3-9. CELL If the Transport Accessory is installed, the Auto Cell Holder cell number that is in the beam is displayed. If the Transport Accessory is not installed, "N/A" is displayed. 3.6 Window Menu Bars Across the top of analysis windows is a menu bar specific to the window. ‘This is referred to in the manual as a window menu bar or simply a menu bar. The following are some of the commands that typically appear in the menu bar. 3:12 . ‘Operating Information Take a sample reading. In some modes, and with some sampling accessories, this command causes the window shown in Figure 3-9 to be displayed. Sample readings are started when [START] is clicked on and are stopped when [QUIT] is clicked on. Insert sanp lets) Click on START when ready Click on QUIT te stop START Figure 3-9. Read Samples Window In most cases, clicking on the right mouse button, with the cursor in any position, performs the same function as and (START]. Most operating modes have more than one analysis window. The other windows are displayed by clicking on the window name, displayed in the menu bar. Display the Method window to select a stored method or create a new method. Clear all data from the window, without leaving the analysis mode. Before the data are removed, the Save Clear window, Figure 3-10, is displayed, allowing the data to be stored, To store the data, click on the box to darken it, verify that the desired file name is displayed and click on [OK]. To change the file name, click on it to display the Results File Directory window, which is described in section 3.9. To change the storage location, click on the displayed location to toggle between [A:\] and [Ba]. fl) Save results before clearing? Results file name: LA: X1008K RES Figure 3-10. Save Clear Window Operating Information 343NOTICE The user can select to have the save box darkened automatically each time the Quit window is displayed. This selection is made on the User Interface window in the Configuration mode, which is described in section 2.4, Print the data displayed on the window. Alphanumeric data are printed on the Dot Matrix Printer. Sample data and other information that are not displayed on the screen because of insufficient room are printed. If the X-Y Plotter is installed, graphic data can be printed on either the Dot Matrix Printer or the X-¥ Plotter. This is selected in the Printer and Plotter Configuration window. Remove the window. Leave an operating mode and return to the Main window, If the method, standards file or results file has changed, the Quit window, Figure 3-11, is-displayed. To store any of the information, click on the associated box to darken it, verify that the desired file name is dis- played and click on [OK]. To change a method file name, click on it to display the alphanumeric keypad and input the desired file name. To change a data file name, click on it to display the Results File Directory window, which is described in section 3.9. To change the storage location, click on the displayed location to toggle between [A:\] and [B:\j. fH) Save changes in the method method 2 Method file name: CA:\IDEFAULT Gl Save results before clearing? Results file name: [CA:\]WORK_RES [on Figure 3-11. Quit Window NOTICE The user can select to have the save boxes darkened automatically each time the Quit window is displayed. This selection is made on the User Interface window in the Configuration mode, which is described in section 2.4. 3-14 Operating Information3.7 Parameter Input | Parameters that pertain to a particular analysis window are listed near the top of the window. The Method window contains a complete listing of analysis parameters. The selected value can be changed for any parameter where the displayed value is a different cofor than the parameter. For example: the parameter, "Wavelength", is a different color that the value, "500". | There are several types of parameter input. These include: Numeric Input When a parameter is clicked on that requires a numeric input, the numeric keypad, Figure 3-12, is displayed. The limits of the input are displayed at the bottom of the keypad. Input the desired value and click on [OK] to accept the value and remove the numeric keypad, Ex- amples of parameters that require numeric input include the wavelength, number of samples, and read average time. Figure 3-12, Numeric Keypad As an alternative, the optional keyboard can be used to input numbers. When the numbers are input, they are displayed in the box at the top of the keypad. When the desired number(s) are displayed, press enter on the keyboard (or click on [OK] on the keypad) to accept the value and remove the keypad, Alphanumeric Input When a parameter is clicked on that requires an alphanumeric input, the alphanumeric keypad, Figure 3-13, is displayed. The maximum number of characters that can be input is indicated by the space provided for the input. Input the desired information and click on | : i : L Operating Information 35[OK] to accept the information and remove the alphanumeric keypad. Examples of parameters that require alphanumeric input include file names, concentration units and sample identifications. Results2 SHBEReEL BKSP CLEAR Figure 3-13. Alphanumeric Keypad As an alternative, the optional keyboard can be used to input this information. When the characters are input, they are displayed in the box at the top of the keypad. When the desired characters are displayed, press enter on the keyboard (or click on [OK] on the keypad) to accept the information and remove the keypad. Option Selection If a parameter has a variety of options, a selection window is displayed when the parameter is clicked on. In some cases, the word "VIEW" is displayed and clicked on to get the selection window. A typical L selection window is shown in Figure 3-14. | O None Ed One cell [Single Cell] OF Auto smpir Number of cells: 6 4 Batch smplr Figure 3-14, Sampling Device Window | | i [OK] ‘ 3:16 Operating InformationEach of the options on the selection window is preceded by a box. Click on the box to darken it to select the desired option. A second click on the box removes the darkened area. Some selection windows accept only one choice. Others allow multiple selections. When the selection(s) are made, click on [OK] to input the option and remove the selection window. Bracketed Options Some parameters require a selection from only two or three options, such as "yes" or "no". ‘The selection for these types of parameters is displayed with brackets, such as [Yes]. When the selection in brackets is clicked on, the selection changes to the other option. For example, click on [Yes] to display [No]. Examples of parameters that have bracketed options include the selection of absorbance or transmittance readings and the use of a background wavelength. Parameters that Cannot Change If the selected value for an analysis parameter is displayed in the same color as the parameter, the value cannot be changed. There are several reasons that a parameter cannot be changed. They include: 1. The method in use has been protected. After a method is developed, it can be protected, so that the analysis must be done with the selected parameters. Parameters used for sample collection must be the same as for the related standards. For example, if the standard curve was calculated using an analytical wavelength of 500 nm, the wavelength for the sample analysis must be 500 nm. Data collection is complete. For example, the temperature cannot be changed if data were previously collected. Operating Information BAT | | |3.8 Method Development and Use Parameter setup for analyses that are repeated often can be simplified by storing the analysis parameters in a method file. Each analysis mode (with the exception of the Fraction Read mode) has a Method window associated with it. The Method window is used to setup, store and retrieve analysis parameters. A typical Method window is shown in Figure 3-15. Each Method window is divided into two parts. The top part is the Method File Directory; the bottom part lists the analysis parameters for the "Method Create BENScAN Save Reni Delete Protect Print Nethog in use: as\DEFA Mole: This is an deanple user note, Read Node: [Abs] Start easier ens 200, am <8 [sec] inits: 3.0008 fmits? G:0998 ar Auvopeints cua ] autosave: (uo J autosave File nane: CAz\ISCANS Function: VIEU Overlay scans Saal in ‘ce: None Scan speed 1260 ne/ain Figure 3-15. ‘Typical Method Window in use". Creating a Method To create a new method: 1 3-18 With the Main window displayed, click on the desired analysis mode. The first window for the analysis mode is displayed. Click on to display the Method window for the analysis ‘ mode. The analysis parameters are listed in the lower part of the window, Operating Information3. If the method to be created is similar to an existing method, click on the existing method name in the directory at the top of the Method window. The existing parameters are displayed on the lower part of the | window. i 4. Click on to display the Create window, Figure 3-16. New File name: CA:\]DEFAULT CANCEL OK Figure 3-16. Create Window Input the new method file name and location in this window. a. To input the location, click on the displayed location to toggle between [A:\J and [B:\]. b. To input the new file name, click on the name that is displayed | following "New file name" to display the alphanumeric keypad. | Input the new name, then click on [OK] to remove the keypad. | c. Click on [OK] to remove the Create window and accept the new file name. d. The new method file name is displayed following "Method in use". 5, To input comments about the method, click on "Note" and use the alphanumeric keypad to input a message with a maximum of 40 characters, 6. To input the analysis parameters, click on the displayed value(s) and | input the desired value(s). Detailed instructions for parameter input are provided in section 3.7. i 7. After the desired parameters are displayed on the Method window, the method can be used or stored, and if stored, protected. To use or to store the method, click on . The method is stored automatically and the appropriate analysis window is displayed with the parameters for the method. To store the method without removing the Method window, click on . This allows more than one method to be created without leaving the Method window. Operating Information 3-19Protected Methods After a method has been developed and stored, it can be protected. The parameters for a method that has been protected cannot be changed, as long as the protection remains. To protect a method: 1. Display the desired method name on the Method window, following “Method in use". 2. Click on to display the Set Protection window, Figure 3-17. SET protection on method: A:\FIXED User nane: Password: Figure 3-17. Set Protection Window 3. Input the assigned "User name" and "Password", then click on [OK]. If both are accepted, the protection is implemented and the window is removed from the display. **PROTECTED** is displayed following the method name on the Method window. To remove protection: Protection is removed using the same steps. If a protected method is displayed following "Method in use", when is clicked on, the Remove Protection window shown in Figure 3-18, is displayed, To delete the protection, input the assigned "User name" and "Password", and click on {OK}. REMOVE protection on method: A:\FIXED User name: Password: CANCEL fo] Figure 3-18, Remove Protection Window 3-20 Operating InformationPlacing the Method Name on the Main Window The method can be listed on the Main window in the area entitled “Custom Applications". To place it there, use the File Utilities mode to copy the method file from the appropriate method directory 10 the "CUST_APP" directory. Up to 30 methods can be listed on the Main window. Using a Stored Method To use a stored method listed in the “Custom Applications" area on the Main window, click on the desired method name. The appropriate analysis window with the values for the method is displayed. NOTICE When a method listed in the "Custom Applications" area is selected, the parameters can be modified on the applications window or the Method window. However, the changes cannot be stored. To store the changes, recall the method from the applications mode, make the changes, store the method, then move it to the Custom Applications directory. To use a stored method not listed on the Main window: 1. With the Main window displayed, click on the desired analysis mode. The first window for the analysis mode is displayed. 2. Click on 1o display the Method window for the analysis mode. 3. The Method File Directory for the analysis mode is displayed at the top of the Method window. Click on the desired method name. The parameters are displayed on the lower part of the window. 4. If the method is protected, the protection is indicated to the right of the method name on the lower part of the window. If the method is not protected, any of the parameters can be modified. 5. Click on to display the appropriate analysis window with the values for the method. Operating Information 321Renaming a Method File 1. Display the desired method name, following "Method in use", on the Method window. 2. Click on to display the Rename window, Figure 3-19. Current file name: AS\FIXED New File name: TEST [eaNcet] [ok Figure 3-19, Rename Window 3. Click on the name following "New file name" and input the desired file name. Then click on [OK] to rename the file and remove the window from the display. 4. The file is stored with the new name when either or (to remove the Method window) is clicked on. Deleting a Method File 1. Display the desired method name, following "Method in use", on the Method window. 2. Click on to display the Delete window, Figure 3-20. WARNING File AZ\FIXED will be deleted CANCEL Figure 3-20. Delete Window 3. Verify that the desired file name is displayed, then click on [OK] to delete the file and remove the window from the display. 3-22 Operating Information3.9 Stored Data The instrument has the capability of storing the data it collects either in internal memory (drive A) or on the optional external disk drive (drive B). Data from most modes are stored in “Results Files". However, some modes have special types of data files, such as "Standard Files" in the protein analysis mode. All types of data files operate the same. Data files are renamed, copied to another location, moved to another location and deleted using the File Utilities mode. Instructions are provided in section 8. The following options are available for data storage: 1. Not storing the data. The data are removed from the window by clicking on either or to display the appropriate window. The data are deleted if the box next to the file name is not darkened. The Save Clear window is shown in Figure 3-10. The Quit window is shown in Figure 3-11. 2. Designating a file name for data storage before the data are collected. The file name is input by clicking on "Results file", displayed with the other parameters on the analysis window. 3, Collect the data and then decide whether to store the data. When the data are removed from the window by clicking on either or , the appropriate window is displayed. The data are stored if the box next to the file name is darkened. Naming a File When "Results file" is clicked on from an analysis window, or when the file name is clicked on from the Save Clear or Quit window, the Results File Directory window specific for the analysis mode is displayed. A typical Results File Directory window is shown in Figure 3-21. To name a new results file: 1. To input the location, click on the location that is displayed following "Selected file” to toggle between [Ad] and [B:\]. . 2. To input the new file name, click on the name that is displayed following "Selected file" to display the alphanumeric keypad. Operating Information 3.233. 4. 5. Aro Resuits File Dirertory Selected File: [Az\]WUORK_RES As 441389 AZNEXAMPLES AS\FIXEDI Figure 3-21. Results File Directory Window Input the new file name, then click on [OK] to remove the keypad. Click on [OK] to remove the window and accept the new file name. The new file name is displayed following “Results file". Recalling Data Data files that are stored in the instrument can be recalled in the mode where they were created. The stored information can be used to recalculate results in the same way that the data was manipulated after it was collected. With the exception of the Fraction Read mode, data cannot be added to an existing file, To recall data: 1 3.24 With the Main window displayed, click on the desired analysis mode. The first window for the analysis mode is displayed. Verify that no data are displayed on the window. If data are displayed, click on . Click on the file name following "Results file". The Results File Directory window is displayed. Click on the desired file name, listed in the directory to place the name after "Results file name”. Click on [OK] 10 remove the Results File | Directory window and display the data in the appropriate analysis window. Operating InformationSECTION FOUR GETTING STARTED The DU Series 600 Spectrophotometer user interface operates on the principle of windows. The "mouse" is used to position an arrow on the window. When the arrow points to the desired position, the left button on the mouse is pressed to initiate the desired action. In these instructions, the positioning of the arrow and pressing the left mouse bution is called “clicking on". 4.1 Power Up Power Up Diagnostics Window When the DU Series 600 Spectrophotometer is powered up, the Power Up Diagnostics window, Figure 4-1, is displayed. If all tests passed, use the mouse to move the arrow so that it points to "Quit", located near the top right-hand corner of the window, and press the left mouse button. Computer and Harduare Diagnostics: Passed PROM RAM Controller Ran Video Control ter Video R Vides Felet RS259 pores T ana 2 EE PROM Passed Spectrophotometer and Systeas Diagnostics: PROM Option Passed Software Option Ran Gption Keyboard Processor tector a Ufetble Lanp Ligne Pat FEES rector Wavelength Drive Systea Clock Passed Figure 4-1. Power Up Window Getting Started 41 |NOTICE If any of these tests fail, refer to the Troubleshooting instructions in section 11.1 of this manual. Programmability is an option and may not be installed on all instruments. The "Quit" command is located in the menu bar at the top of the Power Up Diagnostics window. Most windows have a menu bar associated with them. Commands in the menu bar at the top of a window are referred to in these instructions with single angle brackets, ic. . Main Window When the Power Up Diagnostics Window is removed, the Main window, Figure 4-2, is displayed. The desired operating mode is selected from the Main window. The Fixed Wavelength, Wavelength Scan and Kinetics/Time modes are standard on all instruments. All other modes are optional, and are only displayed if they are installed on the instrument. PROGRAR REROTE CONTROL SCAN AREA GEL SCAN MOLECULAR WEIGHTS 1 DATE _TeRF CELT Prescrn 8.8813 Abs i2/3e/s1 WAAL Figure 4-2. Main Window 42 Getting StartedSources ‘The commands to turn the sources on and off are located in the Permanent Menu bar, which is always located at the bottom of the display. Commands in the permanent menu bar are referred to in the instructions with double angie brackets, i.e. <>. ‘To turn on the visible source, click on <> to display <>. The visible source lights immediately. To turn on the UV source, click on <> to display <>. The UV source requires about 30 seconds to warm up before it lights. When the source lights the command is changed to <>. Do not blank while < > is displayed. ‘The instrument should be allowed to warm up for at least 30 minutes before blanking and taking sample readings. Any reading taken with the sources turned off is invalid. To turn off the sources, click on <> and/or <>. When the source is turned back on, a new blank will be required. Data Collection Modes The DU Series 600 Spectrophotometer has five data collection modes: RediRead™ Mode, RediScan™ Mode, Fixed Wavelength, Wavelength Scan and Kinetics/Time. They are described in the following sections. Getting Started 43 : i i | 1 i42 RediRead™ Mode The RediRead window is used to take fixed wavelengths readings at one or more wavelengths quickly and easily. This window can be displayed whenever the instrument is not collecting data, regardless of the operating mode of the instrument. Data collected in this mode cannot be stored. 1. Click on <>, located in the permanent menu bar at the bottom of the display, to display the RediRead window, Figure 4-3. ReadSanple 500.dnn 6.1278 A Read avg times 0.50 Reac Mode: CAbs) Sample Wavelength ding Re 560 .8nm 8. 500 .Ona 8. a. 8. §00.0nn 569 .Onm Figure 4-3. RediRead Window 2. Set the parameters: a. Click on the wavelength value displayed and input the desired wavelength. b. Click on "Read avg time” and input the desired read average time. c. Verify that the desired reading mode is displayed, [Abs] or [%T]. Click on the mode to change it. 3. Place a cuvette of solvent in the cell holder and click on . (If the instrument has previously been blanked at the selected wavelength using <>, it is not necessary to blank in the RediRead mode. in the RediRead mode does | not affect the blank stored using < >.) 4. Place a cuvette of sample_solution in the cell holder and click on . The reading is displayed in the table on the window. 44 Getting Started5. Repeat step 4 for all the samples. The parameters input in step 2 can be changed at any time. Readings from 11 samples are displayed on the window. When the sample 12 is read, the data is written over the data for sample 1. 6. To print the window, click on . Only the data that are displayed are printed. 7. To remove the RediRead window, click on . | Getting Started 4s43 RediScan™ Mode The RediScan window is used to make a wavelength scan at 1200 nm/min on a sample with minimum parameter setup. Data collected using this window cannot be stored; the Wavelength Scan mode must be used for data storage. 1. Click on <>, located in the permanent menu bar at the bottom of the display, to display the RediScan window, Figure 4-4. Wavelength (na) Figure 4-4. RediScan Window Verify that the proper ordinate label is displayed, [Abs] or [%T]. Click on the label to change it. Verify that the desired wavelength limits are displayed. To change them, click on the displayed value and input the desired value. The sample will be scanned over the displayed wavelength range, only. Place a cuvette of solvent in the cell holder and click on . (if the instrument has previously been blanked in the Wavelength Scan mode at 1200 nm/min over the selected range, it is not necessary to blank in the RediScan made.) Place a cuvette of sample solution in the cell holder and click on . The scan data is displayed. Getting Started | | | |6. The following functions are available to reformat the data: a. The data can be autoscaled by clicking on . b. Individual axis limit values can be changed by clicking on them and inputting the desired value. 7. To display the wavelength and ordinate readings at any point in the spectrum, click on . Then move the mouse to the point of interest in the spectrum and click on the center mouse button to place a vertical line on the spectrum. ‘The values at the place where the vertical line is placed are displayed in the lower right-hand side of the window. To move the vertical line to either the right or left, click on the right or left mouse button, respectively. 8. To annotate the data, click on , Then, click on the graph to position a cross and input information from the alphanumeric keypad or keyboard. Up to four annotations can be placed on the graph. The annotations are printed with the window, but are not stored with the data. 9. To print the wavelength scan in the window, click on . 10. Repeat steps 5 to 8 for all the samples. 11. To remove the RediScan window, click on . Getting Started a74.4 Fixed Wavelength The Fixed Wavelength mode is used to collect data from a series of samples at up to 12 wavelengths. The data can be multiplied by user-input factor(s) to calculate a result at each wavelength. Any of the sampling devices can be used to simplify sample handling. Data can be stored for later recall. To select the analysis parameters: 1. With the Main window displayed, click on "FIXED WAVELENGTH" to display the Fixed Wavelength window, Figure 4-5. Paraneters 2 AZ NFIXEDL Kethod ni Read average tine: 0.68 Read mode: (Abs) Sanpling ‘Sample 10 > 380.0 Pry Factor 56.08 © Factor 230.0 Abs Result. Result 58. : 59.2248 9: 86.8482 Figure 4-5. Fixed Wavelength Window 48 Getting Started2, Click on 10 display the Parameters window, Figure 4-6. Factor Units ng/n! ng/nl agen) ng/ml ng/n) ag/at ng/al ng/nt ag/at ngénl ngent ng/m} Figure 4.6. Parameters Window a, Listed in the Parameters window are 12 wavelength values, with a factor and units that correspond to each wavelength. To change any of these values, click on the displayed value to display a keypad. Input the desired value on the keypad, then click on [OK] to accept the input and remove the keypad. b. The fourth column in the Parameters window is the "Use" column. Each wavelength that is to be used in the analysis must have a | "Yes" displayed. If a "No" is displayed for a desired wavelength, i click on the "No" to display a "Yes", : c. When all the desired values are displayed, click on to remove the Parameters window. The input values are immediately displayed on the Fixed Wavelength window. 3. Readings can be taken in either absorbance or transmittance. The selection is displayed following "Read mode" in the parameter listing near the top of the window. To change the read mode, click on the displayed option. \ To take readings: i. Place a cuvette of solvent in the instrument. Click on <>. 2. If desired, click on the next displayed sample number and input up to an 1i-digit alphanumeric sample identification. If a sample identification is not input, the instrument numbers the samples consecutively. Getting Started +Place a cuvette of sample solution in the cell holder and click on . Data from up to 3 wavelengths are displayed at one time. To display data at other selected wavelengths, click on the right and left arrows, located on the right-hand side of the analysis parameters. Repeat steps 2 to 4 until all samples have been read. To print the sample data, click on . When the analysis is complete, click on . To store the method and/or results, click on the displayed file name(s) and input the desired file name(s). Then click on [OK] to store the data and return to the Main window. The complete capabilities of the Fixed Wavelength mode are described in section 5 of this manual. 410 Getting Started4.5 Wavelength Scan The Wavelength Scan mode is used to collect, manipulate and store scan data, To select the analysis parameters: 1. With the Main window displayed, click on "WAVELENGTH SCAN" to display the Wavelength Scan window, Figure 4-7. EB 7 Readsaaples tabulate ¢-xSeans Scatter eta fethod Saveciear Prin fean directory: VEU — Autoprint: (No 1 Rethod rane: Start viz 200) na futosave: {No 1 Autosave na End wiz 800. nm Scans Overlay scans: (Wo 1 Interv: AE\SCANOOL (1208) angle; Sanpling devi: 0.00 tsec] Scan speed 121 Functions: Scan Smoothing: None | i | | Zeon Zoandut_ Trace ~— Function —_—futoscale annotate Print. 2.cno ; | | Wavelength tom) Figure 4-7, Wavelength Scan Window 2. ‘Twelve parameters are listed near the top of the window. a. Locate the "Start wi" and “End wl" parameters. To change the values, click on the displayed value and input the desired values. b. Verify that the following parameters are as follows: Overlay scans: [No] Autoprint: [No] | Autosave: [No] : Scans per sample: 1 | Sampling device: None i Scan speed: 1200 nm/min I If any of the parameters are different than those listed, click on the | displayed value and input the listed value. Getting Started 44dThe ordinate label and limits are displayed on the graphic portion of the window. To change any of these values, click on the displayed value and input the desired value. To take readings: 1. 2. 412 Place a cuvette of solvent in the cell holder. Click on <>. Place a cuvette of sample solution in the cell holder and click on . The following functions are available to reformat the data: Autoscale - Automatically scales the ordinate axis. Limit changes - The limits on cither axis can be changed by clicking on the displayed value and inputting the desired value. Zoom - The "zoom" feature is used to expand any portion of the graph. Click on , then click on two points on the graph to place crosses at the opposite corners of the area to be enlarged. When the second cross is clicked on, the graph is replotted. This can be repeated as often as desired. To return to the original plot, click on , To smooth the data, click on "Smoothing" and select the desired number of points to use for the calculation. If too few points are used, the data may appear to be noisy. If too many points are used, real peaks can be lost. To display a derivative or log scan, peak pick, valley pick and/or point pick, click on to display the Function Selection window. To select a function, click on to darken the box preceding the selection. Then click on to remove the Function selection window from the display. The data are replotted using the selected function(s). To display the wavelength and ordinate readings at any point in the spectrum, click on . Then move the mouse to the point of interest in the spectrum and click on the center mouse button to place a vertical line on the spectrum. The values at the place where the | vertical line is placed are displayed in the lower right-hand side of the window. To move the vertical line to either the right or left, click on the right or left mouse button, respectively. Getting Started10. i. To annotate the data, click on . Then, click on the graph to position a cross and input information from the alphanumeric keypad or keyboard. The amotation is printed with the window, but is not stored with the data. To print the sample data, click on . To store data before scanning another sample, click on . Click on the displayed file name, input the desired file name, then click on [OK]. The data are stored and the graphic area is cleared. To scan additional samples, repeat steps 2 to 9, above. When all the samples have been scanned, click on . To store the method and/or displayed scan, click on the displayed file name(s) and input the desired file name(s). Then click on [OK] to store the data and return to the Main window. The complete capabilities of the Wavelength Scan mode are described in section 6 of this manual. These include selection of different analysis parameters, overlaid scans, repetitive scanning, scatter correction, spectral manipulation (addition, subtraction and multiplication), and net absorbance. Getting Started 41346 Time Drive The Kinetics/Time mode is used to collect, manipulate and store time drive data. This mode is also used to calculate the rate of kinetic reactions. To select the analysis parameters for time drive: 1. With the Main window displayed, click on "KINETICS/TIME" to display the Plotting window, Figure 4-8. Analytical vavelengtn: $09.6 m Beekgraund vavelangtritto J 286.0 ne Sapling dev: huaber ‘OF ‘sanple: Seapl2 assignnent: {31 Interval tine: 3.000 [sect Curve offset: {None) Total bine: 68.50 tsec} Reac average tine: 1.00 sec i \ Resales file: 7 i i Zoon ZoonDut Trace Autoscele Annotate Print Tenpereture: N/A 3.60000 00008 see 60,808 Figure 48. Plotting Window 2. Fourteen parameters are listed near the top of the window. a, Locate “Analytical wavelength". To change the wavelength, click on the displayed value and input the desired wavelength. b. Locate "Interval time", which determines the frequency of data collection. To change the displayed value, click on it and input the desired value. c. Locate "Total time", which determines when data collection is stopped. To change the displayed value, click on it and input the desired value. 414 Getting Started3. d. Verify that the following parameters are as follows: Background wavelength: [No] Sampling device: None Number of samples: 1 Sample assignment: [S] Read average time: 0.5 sec If any of the parameters are different than those listed, click on the displayed value and input the listed value. The absorbance limits are displayed on the graphic portion of the window. To change the limits, click on the displayed vaiue and input the desired value. To take readings: 1. 2. 6. Place a cuvette of solvent in the cell holder. Click on <>. Place the sample in the cell holder and click on . The Read Samples window is displayed. Click on [START]. The data are displayed as they are collected. If no data appear on the graph, the data probably do not fall within the axis limits. After data collection is complete, the following functions are available to reformat the data: Autoscale - Automatically scales the ordinate axis. Limit changes - The limits on either axis can be changed by clicking on the displayed value and inputting the desired value. Zoom - The "zoom" feature is used to expand any portion of the graph. Click on , then click on two points on the graph to place crosses at the opposite corners of the area to be enlarged. When the second cross is clicked on, the graph is replotted. This can be repeated as often as desired. To return to the original plot, click on . To annotate the data, click on . Then, click on the graph to position a cross and input information from the alphanumeric keypad or keyboard. The annotation is printed with the window, but is not stored with the data. To print the sample data, click on . Getting Started 4187. To display the tabulated data, click on . 8. Before analyzing another sample, click on . To store the data, if desired, click on the displayed file name and input the desired file name, then click on [OK]. The data are stored and the graphic area is cleared. 9, To analyze additional samples, repeat steps 2 to 8, above. 10. When all the samples have been analyzed, click on . To store the method and/or displayed data, click on the displayed file name(s) and input the desired file name(s). Then click on [OK] to store the data and return to the Main window. The complete capabilities of the Kinetics/Time mode are described in section 7 of this manual, These include selection of different analysis parameters, analysis of multiple samples, and rate calculations. 416 Getting Started4.7 Recalling Stored Files Method File If a method was stored in one of the operating modes, it can be recalled by clicking on the file name following "Method name", listed with the analysis parameters at the top of the analysis window. When the file name is clicked on, the Method window is displayed. The stored method files are displayed near the top of the window. To select a stored file, click on the file name. The parameters will be listed in the lower portion of the window. Then click on to remove the method window and display the analysis parameters on the analysis window. Results File - Fixed Wavelength and Kinetics/Time If a results file was stored in the Fixed Wavelength or Kinetics/Time mode, it can be recalled by clicking on the file name following "Results file" on the analysis window. When the file name is clicked on, the directory is displayed. ‘To select a stored file, click on the file name, then click on [OK] to remove the directory and display the data on the analysis window. Scan File - Wavelength Scan If a scan file was stored, it can be recalled by clicking on "VIEW", following "Scan directory" on the Wavelength Scan window. When the file name is clicked on, the directory is displayed. To select a stored file, click on the file name, then click on [OK] to remove the directory and display the data on the Wavelength Scan window. Getting Started 4a7SECTION FIVE FIXED WAVELENGTH The Fixed Wavelength mode is used to collect data in either absorbance or transmittance at up to 12 wavelengths. The reading at each wavelength can be multiplied by a user input factor to calculate a final result. 5.1 Calculations ‘The result is calculated using the equation: Result = Reading x Factor, where the reading is in either absorbance or transmittance. The result is a concentration value if the reading is taken in absorbance. i i j | | | NOTICE Use of this mode to calculate concentration requires that the slope of the standard curve is constant and known, and that the y- intercept is zero. Concentration calculations, derived from a standard curve with multiple standards, are possible using the optional Single Component Analysis mode. Fixed Wavelength 5i | 5.2 Parameter Setup Click on "FIXED WAVELENGTH" from the Main window to start the analysis. The Fixed Wavelength window, Figure 5-1, is displayed. The Fixed Wavelength window is used to select analysis parameters, collect sample data and display stored sample data. eedsanples. Rethod Prin Resuits file: az\voR fethog nane= @:\bEFRULT Read average tine: 0.50 feed noaes (abs} Sanpling Govice: None a anaTe TT 5 OEE > 2508 ys Factor iveoo Factor iveo0 Factor i000 bs Result ADs Result «= abs. Re suit agen agra ng7at Figure 5-1, Fixed Wavelength Window Use the Method window to setup the analysis parameters: 1. Click on to display the Method window, Figure 5-2. The Method window is used to setup analysis parameters, recall stored methods and create new methods. General information on method | windows is provided in section 3.8. 2. To recall a stored method, click on the desired method name in the listing at the top of the Method window. 3. The analysis parameters are displayed on the lower part of the Method window. Input the desired analysis parameters: Method in use - This displays the name of the method that has ( i been selected. If the method is protected, **PROTECTED** is displayed following the method name. If the method is protected, the analysis parameters cannot be changed. To input a new method name, click on . 52 Fixed Wavelength