Life science

Single Cell Chemical Imaging and Spectroscopy with O-PTIR

High resolution chemical cell imaging

Cell imaging with co-located O-PTIR and fluorescence microscopy

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

The top left image is an 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 top right image is the brightfield image. Square markers show locations of O-PTIR spectral collection.

On the bottom is the O-PTIR spectra that 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 from the other locations.

Hydrated cell chemical imaging and spectroscopy with high resolution O-PTIR

Cheek cells were deposited in water on thin substrate (350µm CaF2) and measured in counter-propagating mode using a water dipping objective, 40x, 1.0NA.

Single IR wavelength chemical images were collected in minutes and combined to show ratio images. Individual cells were identified through the imaged and subsequent O-PTIR spectra were collected in seconds.

There is no water background compensation and the spectra show little water absorbances. O-PTIR, due to the large heat capacity of water, is fundamentally less sensitive to water than existing direct IR spectroscopy techniques, such as FTIR and QCL imaging systems.

Sub-300nm resolution imaging of fixed cells

Cheek cells were deposited in water on thin (170um) glass coverslip and measured in counter-propagating mode using an oil immersion dipping objective, 100x, 1.3NA.
The single IR wavelength chemical image was collected at 1740cm^-1 (lipid) in minutes at 50nm pixel sizes, even highlighting lipid domains that were ~ 285nm in size.
Spectra were collected from ~300nm spot size in seconds

Submicron amyloid aggregate imaging in neurons

Left; O-PTIR, single frequency ratio image of 1630/1656cm^-1. Shows distribution of beta protein structures with separation of 282nm! Right; O-PTIR spectra from IR image (left) showing spectra on (#1) and off (#2) the beta protein structure. Spectral differences, clearly show the differences in the amide I band, typical of beta sheet structured proteins, despite these two locations only being separated by 282nm!
Published: Oxana Klementieva et al., “Super-resolution infrared imaging of polymorphic amyloid aggregates directly in neurons”, Adv Sci, Adv. Sci. 2020, 1903004

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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