Cary Eclipse Fluorimeter – Basic Instructions
Susan J. Kohler
Jan.19, 2001
The Cary Eclipse fluorimeter is a versatile instrument with a variety of sample accessories and operation modes. This will serve as a brief introduction to its use for recording emission and excitation spectra and for making "simple reads" at one wavelength.
- Setup
Open the sample compartment to see if the proper sampling accessory is installed. For the purposes of these instructions, either the single cell holder or the 4-cell holder will suffice. If a suitable sample holder is not in place, install it before powering on the instrument.
Turn on the power with the switch on the lower right front corner of the instrument.
Turn on the computer and double click on the Cary Eclipse icon on the desktop.
2. Excitation or Emission Spectra.
- 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.
- 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. Then enter the appropriate wavelengths for your experiment. 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
- Next click on the Accessories tab. If you have the 4-cell holder in place, check the Multicell Holder box. Then check one of the four cells. This will be the cell where you will place your cuvette, so remember which one you chose!
- Next 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.
- 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.
- Choose File-> Exit to close this module.
- Simple Reads
- 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.
- 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 are done, you can save the file in Rich Text Format (RTF) and then import your data into Excel.