Aramis Raman Microscope

Staff Contact: Emory Chan (emchan@lbl.gov)

Instrument Location: 67-4218

Safety Points

You must be authorized to work under WPC activity MF-0001 to operate this instrument.
If using liquid nitrogen, you are required to be on the WPC activity MF-0124 for cryogen handling.
The staff contact for this instrument must approve the materials you will be measuring.
Nanoparticles are not allowed in the open air unless bound to a substrate. The staff contact will let you know which fume hood will be appropriate for you to do your sample prep.
The 4218 lab space is safety glasses exempt with the exception of when liquid nitrogen is being poured, in which case the user is required to ensure everyone in the room has their eyes protected with safety glasses.
The materials provided here offer guidance on using the instrument, but you must still coordinate with the staff member listed above to schedule a formal training.

Standard Operating Procedure

This video walks you through the basic operation of the Raman microscope, including start up & shutdown, loading samples, focusing, spectrometer configuration, and taking and saving spectra. This tutorial is required for all new users of the Raman microscope. During remote training, users should follow along with the video while they are at the instrument, first using a silicon standard and then using your own sample.

Calibrating the spectrometer

This video will teach you how to use the Horiba LabSpec 5 software to calibrate the spectrometer of a Horiba Raman microscope. This calibration ensures that spectral features occur at the correct wavenumber/wavelength.

Taking Raman/PL maps

This video will teach you how to use LabSpec 5 software to acquire hyperspectral maps in which Raman or photoluminescence (PL) spectra are acquired for every pixel in an image.

Using the NIR detector

This video will train users how to use the near-infrared detector (800-1700 nm) on the Horiba LabRAM Raman microscope. This video is part of required training that Users ust complete before they are approved to work with the NIR detector.

HORIBA LABRAM ARAMIS

RAMAN MICROSCOPE

QUICK START GUIDE

Overview

1. Turn on instrument (Section 2)

2. Calibrate the desired grating to the laser wavelength and to the 520.7 cm-1

silicon Raman peak. (See p. 247 of the LabSpec manual v2.02 PDF)

3. Put sample in microscope or macro sample chamber (Section 3)

4. Take spectra (Section 4)

5. Map a sample if desired. (Section 5)

6. Turn off lasers.

7. Clean up after yourself!

8. Sign the log book!

Turning the system on/off

Turning ON

1. Fiber optic white light lamp (under laser table)

2. Laser

3. LabSpec software

Turning OFF

1. Turn off lasers & lamps

2. Evaporate all LN2 only if turning off NIR CCD power supply (optional)

3. Other modules do not need to be turned off.

4. Sign log book!

Setting up Macro sample chamber

The Macro sample chamber is used for large samples that do not need to be

placed under a microscope. This is mainly for homogeneous liquid samples

or solid samples.

1. Click the Setup button in the ARAMIS control panel box (see Section 6)

2. In the “Measure Location” box, choose Macro 90 deg for liquids or retro

(180 degrees) for solids.

3. Change the video camera with Menu:Video > Camera Macro to see the

laser spot while in “retro” 180 degree mode. Adjust XYZ knobs until laser

is in center. With the RTD on, rotate the focus wheel to maximize signal.

ARAMIS Raman Microscope Manual p. 3 of 9

Taking Spectra

1. Open the default configuration:

a. On the top icon bar in the middle, click the button called

“Configurator." The button icon looks like a light blue sheet of

notepad with a "+" sign. It's next to the video camera button.

b. Click the "Load" button and load the file path: "My

Documents/Config/Default config.ngp". This will reset the software

to settings familiar for most Raman users.

2. Turn on live camera with the video button on the top tool bar.

Turn on the reflection fiber light.

3. Lower the microscope stage and load your sample.

4. Turn the microscope to the 10x objective

5. Focus using (stiff) coarse knobs on the left side of the microscope

6. Look for fiber bundle reflection. When the image on screen gets brighter, you are getting closer to focus.

7. Fine focus by rotating the joystick knob.

8. Change to desired objective. If using the 100X objective, BE CAREFUL NOT TO CRASH THE OBJECTIVE INTO YOUR SAMPLE. If using the 1064 laser, use the 100x Leica NIR objective. Use the joystick knob to focus so as not to crash the objective.

9. Stop video camera clicking the stop button (upper right)

10.Set values in control panel (lower toolbar, pictured above). See Section 6

for more detailed information.

a. Set laser wavelength. See Section 8 for laser selection

considerations.

b. Set ND filter.

c. Set hole diameter (aperture). 50 μm for high resolution, 200-300 μm

for homogeneous samples (low resolution, better signal).

d. Set spectrometer field to center of desired cm-1 range.

e. Set options box

i. Set grating to appropriate grooves/mm. See Section 9 for gratings details.

ii. Set Objective to appropriate magnification (e.g. 100x) IMPORTANT!!! Scale bars will be incorrect if you forget this step!

iii. Set data name if you want a prefix for your files.

f. Set acquisition times:

i. Set RTD exposure time (s), exposure time (s), and # of samples to average.

11.Set the detector (Vis or NIR) in Menu: Acquisition > Detector >Sensor

g. If you changed the detector, set the shutter switch on the left side of the ARAMIS box to the right shutter.

(Shutter 1/up) Synapse VIS CCD, (Shutter 2/down) NIR IGA Symphony detector.

12.Click on the “Spectrum RTD” button to take a live spectrum.

Adjust focus, ND filter, hole, and acquisition time to optimize signal.

13.If the x-axis units on the spectrum are not correct (e.g., nm instead of cm-1), you can change them in Menu: Options > Units...

14.Click on the extended range button (three humps) to have the detector take a spectrum over a wider wavenumber range

15.Click the Spectrum Acquisition piggy bank icon to acquire a spectrum.

Setting up a mapping experiment

1.Use the video camera and white light to find the area to be mapped. Focus using the objective to be used for Raman mapping. Stop the video acquisition using the STOP button.

2.Define the area/line to be analyzed using the toolbar on the left.

3.Click on the Mapping Properties icon in order to define the mapping parameters.

4. Click on the Mapping Acquisition button in order to acquire the map.

There are 6 neutral filters installed with the optical densities 0.3, 0.6, 1, 2, 3 or 4.

Lasers

Choose the appropriate laser for your sample. Scattering is proportional to 1/(excitation wavelength)4, and the NIR detector is not as sensitive as the visible CCD, so ideally you would want to use the shortest wavelength possible (e.g. the 532 nm laser). Some samples (e.g. organic samples) may exhibit too much absorption or fluorescence at shorter wavelengths, so your next option should be the 785 nm laser. The 1064 nm laser is used mainly for photoluminescence or for extreme cases to avoid absorption or PL.

Gratings

Choose the appropriate grating given the laser wavelength, your desired spectral resolution, and your desired spectral range.For maximum efficiency, one should match the laser wavelength at which the wavelength that the laser was blazed.Increasing the groove density increases the resolution, increases accuracy, and decreases intensity.

Looking for a printable manual to help you take notes? Look no further than here:

Aramis Raman Microscope.pdf

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