Sop-12: Intensity Calibration Of The Ramanmicro 300 - PerkinElmer RamanMicro 300 Series Getting Started Manual

86 . RamanMicro 300 Getting Started Guide

SOP-12: Intensity Calibration of the RamanMicro 300

The intensity calibration must be performed using the National Institute of Standards and
Technology (NIST) Standard Reference Material. NIST SRM 2241 is the relative intensity
correction standard for Raman spectroscopy when using 785 nm excitation.
Intensity calibration should typically be performed on an annual basis, that is during an
annual instrument service, but also whenever the instrument is moved or following a service
repair.
NOTE: As intensity calibration is not required on a regular basis, NIST Standards are not
included in the basic RamanMicro 300 package. Standard Reference Materials can be
purchased from PerkinElmer (L1321831). Your instrument will be intensity calibrated
prior to shipping and during installation.
1. Ensure the RamanMicro 300 is switched on, if not follow SOP-1.
2. Select the 50x objective lens on the microscope.
3. Ensure that sample NIST 2241 is on the test plate, with the dull surface facing upwards.
4. Open the Scan and Instrument Setup dialog, and on the Hardware tab select
Microscope 50x from the Beam path drop-down list.
5. Ensure that the laser power is set to 100%.
6. Click Spectral Preview and then focus on the NIST standard.
When the NIST standard is optimally focused, close the dialog by clicking Exit.
7. Select the Dark Field (DF) switch position on the Olympus illuminator module.
8. Select Validate and then Instrument from the Instrument menu in Spectrum.
The Validation / Calibration dialog is displayed.
9. Select the Calibration tab.
10. Select Intensity Calibration Y-axis, and then click OK.
The instrument will now perform the intensity calibration routine.
After completing the intensity calibration routine, a report is generated on-screen, which
gives details of the outcome of the intensity calibration.
In the unlikely event that the intensity calibration routine should fail, please ensure the
NIST sample is correctly positioned, and that the laser key switch is set to on (ARM). If
this is the case shut down the PC, switch the instrument off, restart the PC and switch
the instrument back on and repeat the calibration.
11. Repeat steps 2–9 for each objective lens.
12. Remove the intensity correction standard and store safely.
Any data acquired will now be intensity corrected.