Mid Infrared Spectromicroscopy (Mid-IR)

Agilent Operation

Agilent Instrument User Guide

Instrument Orientation

FPA Detector and Liquid Nitrogen

The FPA (imaging) detector is cryogenically cooled for operation with liquid nitrogen. A single detector fill will last for approximately 8 hours. It is very important not to allow the detector to warm up while it is powered on.

Liquid nitrogen filling is handled by a Norhof auto-filler, so you do not need to be present at the beamline to refill the detector. The auto-filler operates on a 4 hour refill cycle. It is possible to trigger an early detector fill to accommodate your experimental schedule. Take note of detector temperature and check the status of the Norhof auto-filler at regular intervals during your experiment. At the end of your beamtime, beamline staff will direct you how to leave the detector / auto-filler. If in doubt, keep the auto-filler running.

Spectrometer and Microscope

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Cary 670 Spectrometer (right) and Cary 620 Microscope (left).

Microscope controls

Manual and software controls exist for most microscope functions.

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Optics / Accessories

The Cary microscope is equipped with both 15X and 25X magnification objectives, as well as a 4X visible-only objective for sample viewing. There are internal zoom optics available which further increase the magnification by a factor of 5 at the cost of signal intensity at the detector. See the Capabilities section of the Agilent page for more details.

The 15X objective can be configured with a Ge ATR imaging crystal.

The main compartment of the Cary spectrometer can be configured for bulk spectroscopy using standard transmission mounts, a single-bounce Ge ATR or a multi-sample autosampler in both the Mid- and Near-IR.

The purge gas is constantly supplied to the sample but the purge shields must be in place. Ensure the clear plastic top plate and objective collar are in place for at least 15-20 minutes before acquiring data. This will provide adequate time for the purge gas to replace the ambient air in the system after the chamber is closed.

Software

Resolutions Pro (left) is the primary software for instrument control and data acquisition.

The Agilent Wizard (right) software assists with stage control, sample orientation and experiment planning.

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 Experimental Parameters

  • Spectral resolution and objective magnification trade off measurement time per area and S/N with increased spectral/spatial resolution.

  • Number of scans directly trades measurement time for S/N.

  • Pixel aggregation (binning) trades file size and S/N for spatial resolution, with no impact on measurement time.

Experimental Setup

These steps should be completed for you by beamline staff, however you should confirm the flap positions and detector cooling before starting your work. If you are remote, you can only confirm that the auto-filler is running.

  1. Start the Norhof liquid nitrogen auto-filler to cool the FPA using the "Fill FPA" desktop link.

  2. Open microscope input flap between FTIR and microscope.

  3. Ensure there is a light path through the main FTIR compartment to the internal DTGS detector and that the compartment flaps are open. Any of the following configurations will work for imaging measurements:

    1. Pike MIRacle ATR accessory

    2. Multisample wheel (wheel removed)

    3. Empty compartment (with purge cover)

  4. Mode specific setup:
    Transmission Confirm that the installed condenser matches the desired magnification
    ATR Imaging Confirm the ATR crystal mount is installed and the FPA has been resized to 64x64 pixels

  5. Turn on the detector

Resolutions Pro Method Settings

Experimental settings are controlled through the Resolutions Pro Imaging Method Editor. Settings specific to your experiment can be saved to a custom method in the Users folder. After saving a new method, you should close and re-open the Imaging Method Editor window.

Before starting your first measurement, double-check the Imaging Method Editor Settings.

Imaging Method Editor Settings

Page

Setting

Description

Comment

Remote Control

Microscope Configuration

Visible Objective

Objective used to collect Visible images

Setting must reflect objective in use

 

IR Objective

Objective used to collect IR hyperspectral images

Setting must reflect objective in use

 

Mode

Optical configuration, can be Transmission / Reflection / ATR

Changes microscope between Transmission / Reflection optical paths

 

High Magnification

Enable HiMag IR optics (additional 5X)

Adds / removes internal 5X optics from IR optical path

Common Settings

Number of scans

Number of co-added scans

Background is commonly set to match sample

 

Resolution

Spectral resolution desired

Determines time to collect a single scan

 

Scan type

Y-axis output of calculated spectra (Absorbance/Reflectance)

 

 

Scan range

Energy range to store in calculated output

Maximum range (FPA) is 3925 - 850 cm⁻¹, recommended 3900 -- 900 cm⁻¹

Advanced Settings: Collect

Speed

Interferometer mirror velocity

Set to 2.5 kHz for 128x128 FPA

 

Interferogram Sampling Interval

Data sampling interval

Set to 4 for 128x128 FPA

 

Number of Pixels Aggregated

Spatial averaging (binning) during collection

Influences S/N and file size, but not collection time

Advanced Settings: Spectrometer Configuration

IR Source

 

Set to "Rear: MIR Source"

 

Beam splitter

Set to "KBr"

Must reflect installed beam splitter

 

Aperture

 

Set to "Open" for FPA Imaging

 

Beam Attenuator Throughput

 

Set to "100%" for FPA Imaging

Agilent Wizard

The Agilent Wizard assists with sample location and experiment planning. At a minimum, you should ensure the objective setting on the Define Regions tab matches the infrared objective you plan to use. Important note: The Agilent Wizard assists with experiment setup and planning, but does not affect the data acquisition step. All data acquisition is controlled through Resolutions Pro.

The Wizard program may be launched from the Agilent desktop shortcut. Visible orientation images are collected of samples loaded onto the stage mounts. Before starting the IR experiment, we will obtain 4x visible images of each sample and import them into the wizard following this procedure: Agilent 4X Image Overviews. Once 4x images of all samples are collected, we can move on to experimental workflows.

Experimental Workflows

The system is now ready to begin sample measurements.