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Correlative Confocal Microscopy and Raman Spectroscopy

The WITec Alpha 300 RA Raman-AFM-SNOM system provides a state-of-the-art confocal Raman imaging, together with AFM and SNOM capabilities in one system. The system based is on an upright microscope from 10-100x objectives with VIS and NIR spectrometers 450nm-1100nm. It has three excitation wavelengths 488, 532 and 785nm. One can use any other microscopy system in the Core Facilites including Superresolution systems and overlay Raman spectroscopy images for correlative purposes. Examples are provided below.

 

Raman Principle


The software helps analyze the data efficiently to identify peaks and interpret.

Offline full-version of the image analysis software is available in the Yellowstone computer in the Core Facilities. Please fill out the training request form here.


Sample Data

Correlative Superresolution Microscopy and Raman Spectroscopy of Human Kidney Stones

Kidney Stones
 

Sample Data

Correlative High-Resolution Widefield Brightfield Microscopy and Raman Spectroscopy of Human Kidney Stones

The below image composite shows individual color coded Confocal Images of Raman components and corresponding spectra and the merged components overlaid on high resolution color brightfield images at different magnifications.
Kidney stones 2

Correlative Raman and Atomic Force Microscopy

Contact and AC Modes

 correlative raman atomic force microscopy

Measurement Position

Correlative Raman and Scanning Near-Field Optical Microscopy

 

Super-Resolution Microscopy

Superresolution microscopy down to 60 nm can be achieved with the WITec scanning near-field optical microscope (SNOM) alpha300 S. SNOM, as compared to other optical superresolution techniques such as STED or STORM which is not dependent on fluorescent tags or specialized laser excitation sources. The operation of the system is straightforward as various measurement procedures such as a high-speed automatic cantilever approach and adjustment can be controlled through the intuitive software.


TrueSurfaceTM Microscopy

With TrueSurface module Raman spectroscopy on rough, inclined, irregularly shaped and curvy samples are possible as it keeps focus on those contoured surfaces. It uses an advanced optical profilometer integrated with the system that provides simultaneous acquisition of topographic and chemical/molecular information.

Sample Data
 

Figure 5mm2
Surface of a 5mm2 polished concrete without TrueSurface (left) where the sample is focused only at the center and TrueSurface image on the right where the entire sample in focus retrieving the curvature of the sample.

 


Brochure and Manuals (pdfs)

WITec-Alpha300-Brochure

Raman Protocols and Procedures

Raman Auto and Manual

Confocal Protocol

AFM Contact Mode Protocol

SNOM Contact Mode Protocol

Free WITec Offline Software Download link-Please fill out the form at the end of this link to receive a download instructions.

https://www.witec.de/products/accessories/software-witec-suite/
 

Manufacturer: WITec Scientific Instrument and Technology GmbH, Germany

Equipment model: WITEC Alpha 300 RS

Location: Room 124 D, Carl R. Woese Institute for Genomic Biology

Contact Person for Correlative and Confocal Raman: Austin Cyphersmith, 217-244-9654, ajcypher@illinois.edu

Contact Person for AFM and SNOM: Austin Cyphersmith, 217-244-9654, ajcypher@illinois.edu

Training Request: https://www.igb.illinois.edu/corefacilities/training-request