<p>Department of Chemistry, Tufts University</p><p>Cary Eclipse Fluorometer –Basic Operation </p><p>These directions are for a normal wavelength scan of emission or excitation. For further details, or other applications, consult the manual.</p><p>Turn ON</p><p>1. Sign the logbook. 2. Turn on the computer, monitor, and printer. Select (double click) the Cary Eclipse </p><p> group on the desktop , and then select . 3. Turn on main instrument. Self-calibration will start. Progress is indicated in the lower left corner of the window. 4. Insert sample. Fluorometer cells usually have all 4 sides clear. THOSE WITH OPPOSITE SIDES FROSTED WILL NOT WORK.</p><p>Software Operation</p><p>1. Select File/Open Method from the menu to load a previously defined method.</p><p>2. If you don’t have a method, select the button. Select the or button, and enter wavelength limits for excitation and emission, and scan speed. Select the tab, and select PMT Detector voltage. Start with Medium if you are not sure which to choose. Select . 3. If you get an error message “System Error-Single Cell Peltier”, software is set to control temperature, while temperature controller power is off. Turn off software temperature control or turn on temperature controller (See below)</p><p>4. Select to scan. 5. If spectrum intensity is weak, or clipped go back to , and adjust PMT voltage, and any other parameters desired. Higher PMT voltage and larger slits give higher sensitivity.</p><p>6. Select again. 7. Use Graph menu to add, remove, scale and format graphs. 8. Use File/Save as... to save data. File type Data (.FDSW) saves a single scan. File type Batch (.FBSW) saves all displayed data. File type Method (.FGSW) saves only parameters</p><p>Plotting the spectrum</p><p>1. Delete unwanted graphs from the window by selecting the graph, then select Graphs/Remove Graph from the menu.</p><p>Revised 02/24/2012 D. Wilbur Department of Chemistry, Tufts University</p><p>2. Select Edit/Clear Report, then Edit/Create Report. Check desired options. 3. Select File/Print Preview from menu. If OK select File/Print.</p><p>Exporting Data 1. To save data as text, select File/Save as from menu, select Spreadsheet ASCII as file type. 2. To export a graph, right click in the graph, select Copy Graph from the context menu.</p><p>Turn OFF</p><p>1. Select File/Exit. 2. Turn off fluorometer. 3. Turn off temperature controller and unplug pump. 4. Shut down Windows. 5. Turn off computer.</p><p>Variable Temperature Operation</p><p>Turn on 1. Turn on the temperature controller (Power switch is on back panel). 2. Plug in the circulator pump. Confirm that water is circulating.</p><p>Hardware control 1. Set desired temperature, using the up/down arrow buttons below the temperature display. 2. If sample stirring is desired, put a stir bar in the cell, and press and hold Stirring Min/Max buttons for desired speed.</p><p>Software control 1. In window select tab, then select Temperature control check box, and Single Cell Peltier from pull down menu. 2. Enter desired temperature (10-70C), select . The REM LED on the temperature controller should be on. Temperature will be displayed in Status Display window below the spectrum window. 3. Software control, if selected, will override hardware control.</p><p>4. The button will abort software control of temperature 5. If sample stirring is desired, put a stir bar in the cell, and press and hold Stir Min/Max buttons for desired speed. There is no software control of stir speed.</p><p>Turn off 1. Unplug circulating pump 2. Turn off temperature controller</p><p>Revised 02/24/2012 D. Wilbur</p>