mIRage-LS

Sub-500nm O-PTIR with co-located Fluorescence and simultaneous Raman spectroscopy: mIRage-LS, a world first and only

Co-located Fluorescence microscopy and sub-micron O-PTIR spectroscopy

Sub <500nm IR chemical spatial resolution

Fast fluorescence identification and imaging of cells, tissues, bacteria

About mIRage-LS

The mIRage-LS sub-500nm O-PTIR platform revolutionizes life science research by combining co-located O-PTIR and fluorescence microscopy. Fluorescence microscopy provides molecular specificity, while vibrational spectroscopy (IR and Raman) offers spatially resolved macromolecular characterization. The integration of these techniques enables researchers to explore biological samples with unmatched detail.

With the combination of the mIRage-LS sub-500nm O-PTIR resolution, simultaneous Raman and co-located Fluorescence, correlative imaging at scales <500 nm can be achieved. This seamless, registration-free platform synergizes these methods, unlocking deeper molecular insights and enabling transformative discoveries unattainable with either technology alone.

Mirage OPTIR mode image_LS
Co-located fluorescence + sub-micron IR of cells

Co-located fluorescence + submicron IR of cells

Neuroglioma cells were stained with G3BP1 for protein stress granules, DAPI for nucleus and BODIPY for lipids.

Image: RBG overlay widefield epi-fluorescence image with red showing protein stress granules, Blue showing nucleus and Green showing lipids. Square markers show locations of O-PTIR spectral collection.

The right side shows the brightfield image. Square markers show locations of O-PTIR spectral collection.

The O-PTIR spectra was collected in seconds from the marker locations shown in the left and middle panes. Clear spectral differences can be observed, consistent with the targeted sub-cellular features. Of particular note, is the subtle shift in the Amide I band of the protein stress granule indicating a likely different protein secondary structure to the other locations.

mIRage-LS applications

Tissue and cell imaging

  • Single Cells: normal/diseased cell differentiation, drug-cell interactions, intra-cellular (lipid droplet) imaging studies
  • Tissues; cell typing, calcifications, disease state, collagen orientation
  • Bacteria; Bacterial identification at the single cell level, cell metabolism studied with stable isotopic labelling (13C, 15N, Deuterium)
Tissue and cell imaging mIRage-LS
Using fluorescence to localize O-PTIR measurements

mIRage-LS uses co-located fluorescence to localize O-PTIR measurements

An Alzheimer’s disease mouse model brain tissue section was stained with Amytracker 630 to highlight amyloid aggregates, AF488 to highlight proteins and DAPI for the nucleus.

In the figure to the left is shown a brightfield image of the stained sample. In the top right is the RBG composite fluorescence image, which highlights in red/orange the regions of amyloid aggregation. Note how some amyloid aggregates highlighted in the fluorescence image are not readily distinguishable in the brightfield image.

At the bottom is an averaged O-PTIR spectra, from the line profile indicated in the fluorescence image, with spectra averaged on (in blue) and off (in red) the aggregate. The average spectrum of the aggregate shows distinct spectral differences in the amide I band with a significant spectral feature at 1631cm-1, typical of protein beta sheet structures.

This clearly demonstrates the utility of combining fluorescence imaging to highlight regions of amyloid aggregation, some of which cannot be readily seen in brightfield microscopy, with submicrometer O-PTIR spectroscopy which can then provide the molecular compositional information, in this case, being particularly sensitive to protein secondary structure, a characteristic strength of IR spectroscopy.

Application note:

mIRage-LS IR microscopy with co-located fluorescence imaging for life science applications

Webinar:

New sub-500nm IR with simultaneous Raman and co-located fluorescence microscopy

Data sheet:

mIRage-LS, sub-500nm IR multimodal microscope

“The capability of the mIRage system to provide submicron simultaneous IR+Raman is truly unique and offers significant advantages compared to other technologies. Moreover, Photothermal team have been amazing to work with!”

Tanya Hutter, Ph.D
Assistant Professor The University of Texas at Austin

“The mIRage obtains valuable information about the structure of multilayered plastic films and ageing of adhesives not available to conventional FTIR and Raman. O-PTIR is valuable for the development of high-quality plastic recyclables”

Madina Shamsuyeva, Ph.D
Leibnez University Hannover, Germany. Institute for Plastics and Recycling Technology

“Our first user experiment in the field of Alzheimer’s went so well that the researchers are submitting a manuscript to a high impact journal. The OPTIR is a disruptive technology.” 

Ferenc Borondics, Ph.D
Principal Beamline Scientist SOLEIL synchrotron France

Applications

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Sub-500nm Laser Scanning O-PTIR Microscope for High-Speed Imaging. With co-located Fluorescence and Raman spectroscopy.

The mIRage-LS addresses both life science applications and the broad application set of multiuser facilities where the most advanced vibrational spectroscopy needs are required.

mIRage® submicron IR spectroscopy and imaging with FTIR quality data

Optimized for Value. Same high performance as the mIRage platform but leveraged for dedicated O-PTIR only measurements and a focused range of accessories for application flexibility.

Need more information?

Discover how O-PTIR technology can elevate your research or help solve your toughest challenges. Our team are happy to assist and answer your questions.