Instructions for the Alpha300 Raman Confocal Microscope

Instructions for the alpha300 Raman Confocal Microscope.

First, turn on the instrument. The instrument is powered off the extension cords at the bottom. There are two of them and one powers the other. Turn on both switches.

You should hear a beeping sound right away, this is the microscope positioning itself.

Turn on the laser from the on switch at the back then position the key in the on position. It will take a while for the laser to stabilize, wait for the power ok green light to turn on.

Once the laser is stable, you can now open the operating software. The instrument is operated using the Witec Control software on the desktop.

Upon opening the software, the instrument will go through a self check and the events will be displayed on the dialogue box. Each major operation should be checked as successful to ensure the instrument works well. If any of the initialization steps is recorded as unsuccessful, do NOT try to run the instrument, check with the people in charge of the instrument in the lab.

After successful initialization, you are now ready to calibrate it. This is done using a silicon sample and observing the LO phonon line at 522cm-1 . Place the sample on the stage and focus in. As a rule of thumb, always move the microscope Z control up first for safety purposes to avoid crashing the objective. Turn on the illumination, and as you focus look at the sample on the video screen to determine if you are in focus. Once focused, push in the block to close out the white light then pull out the push out rod to allow the laser light to illuminate the sample and the light to be collected back into the spectrophotometer. Click on run oscilloscope to determine if you are getting any peaks. Optimize the counts on the line by focusing in and adjusting the rods on the fiber coupling optical units. If this is your first time to use the instrument consult with a more experienced student before performing this step. Once you are satisfied with the number of CCD counts, stop the oscilloscope mode and remove sample. You are now ready to run your samples.

Now place your sample on the sample stage and focus in once again observing the image on the video screen. Once you determine you are in focus, block the white light and pull out the push rod and run the oscilloscope. If the signal/peaks you are getting satisfy you, then choose what kind of data you aredesiring to get from your sample from the menu on the left hand of the screen.

For single scans: Choose how many scans you want to accumulate, integration time desired and then click on acquire single scan.

Image scans: Choose your integration times, geometry, type of scan and then click on acquire scan. To acquire images you will need prior knowledge of the characteristics peaks of your sample. Creat a filter for these in the filter box and lock it to allow intergration of the peaks for the images. Remember to save your data. These are the two most common runs on the instruments. If you desire more sophisticated runs please consult with more experienced members of the group.

Once you are done with your experiments, make sure you have the microscope raised up and then close out the software. It will take a while since the camera has to warm up. The software will only close once the camera is warmed up. Turn of the laser by turning the key into the off position then the switch at the back.

Shut down the computer and then finally switch off the power plugs at the bottom.