Nano-scale Optical and Hyperspectral Imaging CytoViva, Inc.
Sam Lawrence – Chief Executive Officer
Integrated optical & hyperspectral imaging technology… • Patented high S/N illumination optics, specifically
designed for imaging nano-scale samples
• VNIR hyperspectral imaging system (HSI) enabling
pixel level spectral characterization of the sample being imaged
…specifically designed to advance nano-scale research.
Live oral cancer cells with
aggregating Au nanoparticles
Technology
Overview
VNIR
Spectrophotometer Optical Camera
Optical Image Hyperspectral Data
CytoViva Equipped
Optical
Microscope
Motorized
Stage 150w Halogen Light
Source
CytoViva Dual
Mode
Fluorescence
Module
Current System
Footprint
Functionality Enabling Improved Optical Performance: 1. Pre-aligned Koehler illumination: Precisely focuses
the source light onto the entrance slit of the annular condenser
2. Main feature of critical illumination: Focuses the light precisely on the same plane of the sample as the focal point of the objective; result of pre-aligned Koehler illumination
Patented Optical Illumination System enables nano-scale imaging
Pre-aligned
Koehler Illumination
Critical Illumination
* Optics Letters, Vol. 31, Issue 19, pp. 2855-2857
* Annular illumination produces an improved point spread function. Through design enhancements in the alignment and focus of annular Illumination, CytoViva produces significantly improved optical performance over other comparable techniques including standard darkfield (annular) illumination.
Optical
Imaging
How It Works
• The VNIR spectrophotometer mounts onto a microscope equipped with the CytoViva Optical Illumination System
• It captures the unique reflectance spectra of objects from the microscope field of view (VNIR spectrum from 400nm-1,000nm)
• The complete spectra for each pixel of the
CCD detector is captured (pixel size as small as 128nm)
• Spectral data is reported in high resolution (down to 2.0nm)
• The data is presented as a spectral curve
and as a RGB image
• Detailed quantitative analysis of each object in the field of view can be performed.
Diagram of Diffraction Grating Hyperspectral Imaging System
Hyperspectral
Imaging (HSI)
Comparison of standard darkfield vs. CytoViva Optical Illumination System
Both images represent the identical field of view of 50nm colloidal gold captured under identical conditions: Olympus BX-41, 40X objective, Dage MTX color digital camera at a constant exposure and an X-Cite 120 light source.
CytoViva Optical Illumination System Standard darkfield condenser
50nm AuNP on glass substrate 50nm AuNP on glass substrate
Optical
Imaging
a.) Standard darkfield microscopy
b.) CytoViva Optical Illumination System
S/N Response of CytoViva Optical Illumination System
• Nanoparticle target is
scanned
• Unique spectral signatures of nanoparticle is loaded in a library database
• New sample containing
nanoparticle is scanned
• Using nanoparticle spectral library, nanoparticle is mapped in the new sample
AuNPs mapped (in red )
Library spectra from AuNP
Epithelial cells incubated with AuNPs: hyperspectral image
AuNPs in solution: hyperspectral image
Mapping AuNPs
in live cells
NIST IIT Madras India Dow Chemical
Nanomaterials cell interactions Nanomaterials development Nanoparticle analysis
FDA Taiwan National University Johnson & Johnson
Nanotoxicology Nanomedicine Nanoparticles in composites
NASA Rice University Pfizer Nanocomposites Nanomaterials characterization Pathogen characterization
Argonne National Labs Max Planck Institute Germany Schering-Plough
Nanoparticle development Nanoparticle development Nanoemulsions
NIH-National Cancer Institute University of Sao Paulo Merck
Nano drug characterization Nanomaterials development Nanomedicine NIOSH MD Anderson Cancer Center Proctor & Gamble Nanotoxicology Nanomedicine Emulsions & nanoparticles Lawrence Berkeley Labs Agroscope Zurich Switzerland Novozymes Biofuels development Nanoparticle plant tissue interaction Biofuels development
Sample Clients
and Applications
Nanomaterial Characterization
Examples
200nm Au shells (no PEG) 200nm Au shells (PEG coated) 200nm Au shells (no PEG)
PEG coated NPs are spectrally different from non-PEG coated
Characterizing Drug-Coated
Nanoparticles
AgNPs in Solution
Spectra from individual non-dispersed AgNPs
Aggregating AgNPs in water
Dispersed AgNPs in ionic solution Spectra from individual dispersed AgNPs
Hyperspectral Scan of Toray Fiber Sample #1 No CNT Coating
Hyperspectral Scan of Toray Fiber Sample #3
Specialized GOx CNT Coating
Hyperspectral Scan of Toray Fiber Sample #2
Raw CNT Coating
Hyperspectral measurements of single pixel areas from the three different samples quantitatively confirms different surface chemistry from each sample.
CNT characterization
application
Hyperspectral Image Scan Reference Spectra of GOx coated CNTs
Scanned image and Zoom illustrating GOx coated CNTs adhering to fiber
Overlay image illustrating all areas in red containing the selected spectral profile and location of the GOx functionalized CNTs
CNTs Mapped in Sample
Derived from previously scanned pure GOx coated CNT sample
CNT characterization
application
Nano-Bio Interface
Examples
Hyperspectral image 100x: Algae cell + AgNP
AgNPs in Algae
spectral library combined : Ag from all environments
Mapping in red all pixels matching spectral library Algae cell + AgNP
spectral library: AgNPs inside algae cell: chlorophyll modifies AgNP spectral response
AgNPs in Algae
Spectral signatures collected from the CNT sample and loaded into the
spectral library
Aggregating CNTs on glass slide
MWCNTs inhaled by mouse
in lung tissue
The red areas are the pixels matching the CNT spectral profiles within the spectral library.
Hyperspectral Scanned Image 100X
Carbon Nanotubes inhaled
by mouse in lung tissue
AuNP in Cancer Cells
Hyperspectral image: Rala coated AuNP in cells
Spectral library created from aggregating Rala coated Au particles in cells
Pixels in red indicate presence of aggregating Rala AuNP spectra in cells.
AuNP pixel match classification
Spectral Image: Sample 6
Spectral library
created from
Sample 6
Pixels in red indicate presence of cisplatin in tissue
Mapping bare drug
in tissue
TiO2 Injected in Whole
Animal Organism
Whole animal organism control TiO2 nanoparticles injected in whole animal organisms
TiO2 nanoparticles in solution
Note: This spectra response incorporates characteristics of both samples
CytoViva Inc. is a private company affiliated with Auburn University…
CytoViva technology is utilized in over 300 research laboratories world-wide
U.S.
Canada
Taiwan
Singapore
India
Japan
UK
Korea
Australia South Africa
Global
Presence
Italy Switzerland
Germany
Norway
Netherlands
Brazil
China
CytoViva
3D Optical Imaging
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