UA Biophysics: Spectrofluorometer Instructions
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To get authorization to use it, please send an email to Lab manager.
Before beginning any work on the equipment, make sure it is in good condition. If something does not seem right, should be reported to the lab manager.
TURN ON THE EQUIPMENT:
- Turn off computer and instrument before starting the lamp power supply.
- Remove unnecessary optical filters, polarizers, or another element in the optical pathway.
- Set the current knob of the lamp power supply completely counter-clock-wise turn the knob to the minimum current level. With Laurdan or DPH, use 1 mm excitation slits, 2 mm emission slits, and remove all the grating; while con Calcein, use 0.5 mm slits and install almost 3 grating. This setting may change during measurement.
- Turn on the lamp power supply and wait 10 min to stabilize. Then set the current to 15 -18 A.
- Turn on the circulating chiller using the black power switch, bubbles must move in the hoses.
- Turn on the temperature controller using the power switch on the back panel. Then push the run/stop button. If the temperature doesn’t change when you are manipulating the Vinci software, press the run button again.
- Turn on the instrument, the switch (red) is located on the right rear side of the instrument. You should hear the sound of the shutters and motors.
- Start Vinci software using the shortcut icon on the desktop. The open window is for analysis, for launch the window to measurement software: select <Experiment> and <Experiment and Instrument Control>. Dialog window is displayed, select “Biofisica” settings. Then, a characteristic sound from all the automatized parts is heard while the pass the software test. Please don’t try to scroll or resize Vinci windows, or open other programs because Vinci will crash.
- Check the dark signal in the bottom of the control window. With closed shutters the dark signal is between 300 and 1500 counts/sec.
- Check the monochromator dial wavelengths, if all was fine the last time when you shut down the instrument, it should be at 200 nm. Otherwise, ask for calibration procedure to the lab manager. Check the calibration when no typical intensity is displayed.
- Select <Settings> and <Global Settings> for save experiment file automatically in the output directory.
- Charge an old experiment or setup a new one. Remember that 1.5 ml is the minimum volume in the cuvette.
- Exit Vinci experiment and instrument control. Always, exit Vinci before shutting down the PC1.
- Turn off PC1 and set the lamp power supply to minimal current.
- Stop and Turn off the temperature controller.
- Turn off the circulating chiller
- Wait 10 min, and turn off the lamp power supply.