Cary Eclipse Fluorescence Spectrophotometer - Basic Operating Instructions

January 2004

The Cary Eclipse fluorescence spectrophotometer is useful for obtaining fluorescence excitation and emission spectra, individual fluorescence measurements, and other fluorescence measurements. The operating modes are software-driven applications that the user can select from the main screen. Two applications described in this document are the SCAN and SIMPLE READS applications. Additional information about these and other applications are available in the online HELP for the instrument.

Startup

These instructions assume the use of the basic sample holder which accommodates a single 1-cm cell. A thermostatted cell holder is also available. Note the location of the label on the sample cell. For best precision hen making measurements, always place the cell in the holder with the same orientation.

1. Open the sample compartment to verify that the proper sampling accessory is installed before powering on the instrument.
2. Turn on the power with the switch on the lower right front corner of the instrument.
3. Turn on the computer and double click on the Cary Eclipse icon on the desktop.
4. If you are a new user, click on the Cary Eclipse Tour icon for a brief introduction to the instrument.

Warning. The xenon lamp emits high-energy UV radiation that can cause serious damage to your eyes if viewed directly. The lamp emits only when making measurements. Close the compartment lid when making measurements.

Scan (Excitation or Emission Spectra)

1. Setup
   • Double click on the Scan shortcut in the Cary Eclipse window. The scan window will open and after a minute the stoplight icon should turn green.
   • A small Status Display window shows the current settings. If not visible, select View/Status Display from the main menu.
   • Click on the Setup.. button. A window will open showing you the various setup options. Check the following on the page with the Cary tab:
     • Data Mode: fluorescence
     • Scan setup: choose Emission or Excitation.
     • Enter the appropriate wavelengths for your experiment.
     • Slits: The default values of 5 mm are a good starting place for the excitation and emission slits.
     • Scan Control: medium is a good starting value
     • Click on the Options tab.
       • If you check the Overlay Traces box you will get the spectra overlaid in one window. If the box is unchecked, each spectrum you take will appear in its own window.
   • When you are done, click Okay. The setup window will disappear.
2. Obtain the spectrum.

   Click on the big Start button. Enter a sample name when prompted. The instrument will acquire a spectrum. When it is done, click on the vertical arrow icon and the data will be scaled to fill the window. You can move the cursor along the spectrum and read its coordinates in the lower right corner.
   If you choose Graph ->Remove all traces (or click the trash can icon) the spectra will be removed from the window.

1. Finish

   Select File/Exit to close the application..
   

Simple Reads

1. Setup
• Click on the Simple Reads shortcut in the Cary Eclipse window. The Simple Reads window will open and after a minute the stoplight icon should turn green.
• Click on the Setup.. button. A window will open showing you the various setup options.
  • Set the excitation and emission wavelengths.
  • Note that there are also places to set the excitation and emission slit widths and to change the averaging time. Try the default values of 5 mm slits and 0.10 sec.
• When you are satisfied with the setup, click OK. The setup window will close.
2. Take readings
• Click the Read button. The instrument will take a reading and display it on the screen. Each time you click Read, a new value is recorded.
• When you have completed your readings, you can print a report or save the file in Rich Text Format (RTF) and then import your data into Excel.
3. Finish
   Select File/Exit to close the application.

Shutdown

At the end of the lab period:

• Close the Cary Eclipse software (File/Close).
• Shut down the PC
• Switch main power button on the instrument to OFF.