Life science

High resolution tissue imaging with multi-modal O-PTIR

Breast tissue calcification – Demonstration of <1 micron spatial resolution with O-PTIR

A: Optical image (mosaic). Red box indicates IR image measurement area. B: Single frequency image at 1050cm^-1 to highlight calcification locations. C: O-PTIR Spectra from colored circle markers in IR image (B). IR image area 200×200 microns at 500nm step size. Image time, ~10mins.

Calcification IR image at 1050cm^-1, clearly resolves calcifications averaging only a few microns in size, many even <1 micron. At 1050cm^-1, traditional FTIR has a spatial of ~12microns, which is much larger than the actual features, which is why such small an localized calcifications had not been seen before.

Sample courtesy of Prof Nick Stone, Exeter University, UK. Publication in preparation (Dec, 2020)

Mouse Brain tissue section – Multi-modal, label-free chemical imaging

Label-free chemical imaging is essential in life sciences as it visualizes and characterizes biological samples without dyes, preserving their natural state and minimizing artifacts. It enables the direct identification of molecular composition, offering precise insights into biochemical processes with high specificity.

Shown below is a standard histological thin section of mouse brain, chemically imaged for key macromolecules and metabolites.

Label free imaging in 10’s of minutes with high spatial resolution.

Initial brightfield (BF) and Autofluorescence (AutoFL) images are shown on the left hand side. All images, BF, AutoFL and IR chemical images are inherently perfectly spatially registered as there are no optics (objective) or sample movements between these images.

In red, cell nuclei are clearly highlighted in the image by the 1080/1140cm-1 ratio. In green, creatine is highlighted by the 1040/1655cm-1 ratio and lipid is highlighted by the 1738/1665cm-1 ratio.

Representative single point spectra (~400nm spot size) from each of these chemical images is shown in the bottom pane with location marked by white circles.

Asterisks on spectra indicate the image ratio wavenumber positions described above.

Using 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 right is shown a brightfield image, top left, 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.

Webinars

Life Science

Alzheimer’s Disease Research with Sub-micron Simultaneous IR+Raman: Co-localization of Beta-Sheets and Carotenoids in Aggregates

Two-part webinar to learn how submicron IR (O-PTIR) and Raman has been applied to characterize the co-localization of amyloid β aggregates with carotenoids in human brain tissues to help better understand Alzheimer’s Disease progression.

Life Science

Life science applications using novel submicron simultaneous IR and Raman microscopy – A new paradigm in vibrational spectroscopy

Professor Ji-Xin Cheng of Boston University Photonics Center, author of over 230 peer-reviewed papers, co-inventor of CARS microscopy and pioneer in the field of O-PTIR applied to life science systems, in conjunction with Dr Mustafa Kansiz, Director of Product Management at PSC are proud to present this webinar.

Life Science

Amyloid aggregates in neurons – Life science applications using submicron simultaneous IR and Raman microscopy

Dr Oxana Klementieva and co-workers identifying amyloid-beta aggregates directly in neurons (neurites an dendritic spines) at the subcellular (submicron) level using Optical Photothermal Infrared (O-PTIR) imaging (mIRage submicron Infrared Microscope).

Life Science

Collagen orientation, fiber to submicron fibril life science applications of O-PTIR, from tissues to single cells and bacteria

In this webinar, we will show how, submicron simultaneous Optical-Photothermal IR (O-PTIR) spectroscopy and Raman microscopy (IR+Raman) is being used and published in life science applications, from tissues, to cells and even single bacterial cells.

Life Science

Single and intra-cell bacterial IR spectroscopy

Life science applications of O-PTIR, from tissues to live cells and single cell bacteria. In this webinar, we will show how submicron simultaneous Optical-Photothermal IR (O-PTIR) spectroscopy and Raman microscopy (IR+Raman) is being used and published in life science applications, from tissues, to cells and even single bacterial cells.

Life Science

Live single cell analysis with simultaneous submicron IR+Raman spectroscopy

In this webinar, Prof Peter Gardner (Manchester University, UK) will present results from their recent and first published study on live (and fixed) cell analysis with simultaneous submicron IR+Raman microscopy with wide spectral coverage using two pump lasers (QCL and OPO).

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