Ruby Fluorescence measurement
Laser box details
The laser box has a few important features:
Laser - A 532 nm 80 mW laser which will irradiate your sample. It is a Class 3B laser, so it is not safe to view.
DAC holder - where you need to place your DAC so that it gets irradiated
Optical fiber - this is attached to a spectrometer, for detecting the fluorescence of your sample
Camera - a camera inside the box to see what is going on
Thermocouple - attached to the laser to make sure it doesn’t overheat
Interior light - a light inside the box, switch on and off via the “Interior Light” button on the grey box
Motorised stages - where the laser is mounted. This allows you to remotely move the laser to center it on your sample and get maximum sample
Due to the danger of the laser, it can only be turned on when the box is locked up.
Workflow
Part 1: Laser and Laptop Set up
Move the optical fiber out of the way by moving it along the rail
Put your DAC in the clamp and tighten with the thumb screw
Move the optical fiber back along the rail so it is close to your sample. It does not need to be tight up against it.
Close the red door and lock with the key
Sign into laptop:
Username MXuser
Password Beam109Line
Open the camera viewer:
Open Razor Synapse
Click on Razor Kiyo X to view camera
Turn on the motors for the stages. Each one needs to be turned on, as well as the switch on the base they are both attached to.
Open Kinesis to control the motors. You may need to connect to the motors, but clicking Connect in the top left corner.
Home the motors.
Move the motors to starting positions.
Top motor is the Y axis (up/down). Start at 0.4 mm
Bottom motor is the X axis (left/right). Start at 2.6 mm
Part 2: Fluorescence Measurement Set up
Note: This will need to be done any time after the laptop has gone to sleep.
Open OceanView on the laptop
Select Spectroscopy Application Wizard
Select Fluorescence
Click Next to select Active Acquisition
Change Scans to Average to 10 and click Next
Click the big lightbulb icon at the top to store the dark spectrum
Tick the box for Add clean peaks to schematic
Click Finish
Part 3. Ruby Fluorescence measurement and analysis
Click the Play button. The graph should update every few second and just be showing noise.
Turn on the laser:
Press the Safety Reset button if it is flashing. This allows power to the laser
Turn the Laser ON/OFF switch to ON
Monitor the temperature of the laser via the thermocouple. If it gets above 26 degrees, turn it off and open the door to allow it to cool down.
Click the up-down arrow icon to scale the graph height to fill the window. You should be seeing two peaks.
If the peaks are small, or not present, you will need to adjust the laser position.
Click on the Jog button on the controller, and change Step Size to 0.05 mm for both motors.
Jog each motor slowly to see if the signal increases.
If you still can’t get a signal, talk to your friendly beamline scientist.
Click the Play/Pause button so that the collection stops after a single measurement. You might want to do it a few times to get a good looking peak.
Right click on the graph and a blue line will appear with a readout of the wavelength under the graph. This is a good way to find your max peak, or easily see if the fluorescence peak has moved since your last data collection. You can also zoom in on a selected section of the graph by clicking the icon that looks like a magnifying glass with a 2 inside it. Note down the wavelength of the peak of the second doublet.
Open the Excel spreadsheet Ruby fluorescence measurement which is saved on the desktop. Make a folder under your name and save the spreadsheet under a new name for the relevant pressure and experiment.
Enter the wavelength found in Step 6 into the bright green cell called Lambda R1.
The pressure in the cell is given in column C. Save the spreadsheet again and close.