SBIR-STTR Award

This proposal will lead to a Breakthrough sub?cellular 3D chemical composition research platform for live cells, based on the new invention of sub?micron confocal photothermal IR micro?spectroscopy (CPIR). Label?free sub?micron chemical imaging and spectroscopy has long been sought for visualization of biomolecules and materials in complex living systems. Many diseases first manifest themselves at the cellular, or subcellular level. IR micro?spectroscopy is a powerful technique but its use in life sciences in general and cellular analysis in particular has been limited due to the following 2 key limitations: a) Spatial resolution limited by diffraction to ~ 10 µm. b) Inability for in?vivo imaging due to strong IR absorption by water. This proposal eliminates both of the above limitations and is based on a recently published patent? pending breakthroughby the PI, Prof. Ji?Xin Cheng of Boston University and is licensed exclusively by Anasys Instruments for commercialization. Prof. Cheng is one of the world's leading vibrational spectroscopy researchers and is the co?inventor of the CARS (Coherent Anti?Stokes Raman Scattering) microscope, which is a major innovation in the field of Raman Spectroscopy. Anasys pioneered the field of AFM (Atomic force microscope) based photothermal nanoscale IR Spectroscopy where an AFM probe detects the photothermal signal induced by IR absorption. The goal of this proposal is to create a high speed confocal optical microscopy platform capable of 300 nm IR Spectroscopy in fluid. This platform can be used in sub?cellular chemical composition research of live cells which has many applications in the life sciences. The proposed CPIR platform is highly innovative because it would enable real?time imaging of lipid metabolites in single live tumor cells based on fingerprint IR bands. Compared to the widely studied genetic aspect of cancer and the well?known Warburg effect, appreciation of the role of lipids in cancer development is still emerging. Aberrant expressions of lipogenic genes have been found in brain, mammary, prostate and many other cancer. Even with these discoveries, lipid metabolism has not been used as a prognostic factor for cancer aggressiveness due to lack of differential detection and quantitation technology. Our proposal aims to fill this gap by quantifying the amount and composition of lipid droplets, an important aspect of lipogenesis in tumor cells. Though we focus on lipid metabolites here, our spectroscopic imaging platform is generally applicable to monitor the intracellular dynamics of other metabolites, anti?cancer drugs, and nutrition molecules such as fatty acids and amino acids, thus having a far?reaching impact on cancer research.

Public Health Relevance Statement:
Project Narrative The goal of this proposal is to create a sub?cellular 3D chemical composition research platform for live cells, based on the new invention of sub?micron ? confocal photothermal IR micro?spectroscopy. It will a)Provide 300nm spatial resolution for IR imaging and Spectroscopy at high speeds. b) Work in fluid.

Project Terms:
absorption; Ache; Amino Acids; anticancer research; Antineoplastic Agents; base; Biological; Biological Sciences; Boston; Brain; cancer cell; cancer imaging; Cells; Characteristics; Chemicals; Cholesterol Esters; commercialization; Complex; Detection; Development; Disease; experimental study; Fatty Acids; Fingerprint; Genes; Genetic study; Goals; Image; Imagery; imaging platform; improved; in vivo imaging; innovation; instrument; interest; invention; Label; Lasers; Legal patent; Letters; leukemia; Lighting; lipid biosynthesis; lipid metabolism; Lipids; Lipoproteins; Liquid substance; live cell imaging; malignant breast neoplasm; malignant mouth neoplasm; Malignant neoplasm of brain; Malignant neoplasm of prostate; Malignant Neoplasms; Mammary gland; Maps; Measures; Metabolic; Metabolic Pathway; Metabolism; Microscope; Microscopy; Monitor; Morphologic artifacts; nanoscale; neoplastic cell; nutrition; Optics; Organism; Performance; Population; Prognostic Factor; Prostate; Publishing; Raman Spectrum Analysis; Renal carcinoma; Research; Research Personnel; Resolution; Role; Sampling; Scanning Probe Microscopes; Signal Transduction; simulation; Small Business Technology Transfer Research; spectroscopic imaging; Spectrum Analysis; Speed; submicron; System; targeted treatment; Techniques; Technology; therapy development; Time; Tissues; tool; Triglycerides; Universities; vibration; Warburg Effect; Water; Work

Optical photothermal IR (OPTIR) microscopy for chemical imaging of living cells at sub-micron resolution Project Summary/Abstract Photothermal Spectroscopy Corp (PSC) and Prof. Ji-Xin Cheng of Boston University in collaboration with Prof. Rohith Reddy (University of Houston) propose to develop, validate, and commercialize a novel technical called Optical Photothermal Infrared (OPTIR) spectroscopy. This optical microscope-based instrument that will provide microscopic chemical analysis and chemical imaging for the life sciences with sub-micron spatial resolution. The new OPTIR technique is based on infrared (IR) spectroscopy, one on the most commonly used analytical techniques for chemical analysis. IR spectroscopy has already been applied to diverse research problems in the biomedical sciences, but it has not been widely applied for live cell research because of two key limitations. First, fundamental limits on spatial resolution (~3 – 30 µm) have prevented broad application of IR microscopy to single-cell and sub-cellular investigations. Second, strong IR absorption by water has dramatically hindered application of IR spectroscopy to live cells and other hydrated samples. Leveraging successful Phase I research, this proposal aims to overcome the key barriers of conventional IR spectroscopy, while also providing ~10X better spatial resolution. This project will involve two major thrusts: (1) the development of a commercial OPTIR microscope that is designed specifically for the life sciences research community; (2) demonstration and validation of the OPTIR technology on specific problems in cancer research. Commercial instrument development will be performed at PSC facilities in Santa Barbara CA in collaboration with the group of Prof. Ji-Xin Cheng at Boston University. The cancer related research will be performed at both Boston University and the University of Houston. Instrument development activities will include the development of a fully engineering OTPIR prototype for the life sciences community, focus on improvements in spatial resolution, measurement speed, ease of use, and compatibility with measurements of live cells and other hydrated samples in physiological environments. Cancer related research will focus on using the OPTIR technique to measure and map key spectroscopic markers that are indicators of cancerous cells and cancer aggressiveness. Further studies will investigate correlations between IR spectroscopic markers and tumor fighting efficacy. The project team is uniquely qualified to perform this work. PI Prof. Cheng is an award-winning research pioneer in development and application of novel spectroscopic and imaging techniques for the life sciences while the technical and commercial team at PSC has decades of successful experience developing and commercializing high resolution imaging and analytical instrumentation. Project collaborator Prof. Reddy is an award winning researcher with extensive expertise in the application of infrared spectroscopy to issues in cancer research. Successful completion of this project will lead to a next-generation single-cell analysis tool that will profoundly impact cellular and sub-cellular biology by providing functional information about changes in cellular state, including membrane order, protein folding, and DNA damage. These changes can be measured in live cells and over time to provide new insights into processes in health and disease.

Public Health Relevance Statement:
Optical photothermal IR (OPTIR) microscopy for chemical imaging of living cells at sub-micron resolution Project narrative This STTR project will develop a new optical microscope-based platform that performs label-free chemical analysis with sub-cellular spatial resolution using a novel form of high resolution infrared spectroscopy. This instrument will enable chemical analysis on life sciences samples including tissue and live cells with sub-micron spatial resolution via optical photothermal infrared (OPTIR) spectroscopy. This next-generation single-cell analysis tool and its ability to measure and map cancer related biomolecules on a sub-cellular level will profoundly impact cancer research, impacting fundamental understandings of cancer progression and treatment.

Project Terms:
absorption; Adoption; anticancer research; Area; Atlas of Cancer Mortality in the United States; Award; base; Biological; Biological Sciences; Boston; Cancer Biology; cancer immunotherapy; cancer therapy; Cancerous; Cells; Cellular biology; Chemicals; chimeric antigen receptor T cells; Cholesterol Esters; Collaborations; Communities; Computer software; design; Detection; detector; Development; Device or Instrument Development; Disease; DNA Damage; Drug resistance; Engineering; Environment; experience; fatty acid metabolism; fighting; Health; Heating; high resolution imaging; Hydration status; Image; imaging system; Imaging Techniques; improved; infrared microscopy; Infrared Rays; infrared spectroscopy; insight; instrument; instrumentation; interest; Investigation; Label; Lasers; Light; light scattering; Lighting; live cell imaging; Malignant neoplasm of ovary; Malignant neoplasm of prostate; Malignant Neoplasms; Maps; Measurement; Measures; Membrane; Metabolic; metabolic imaging; Methods; Microscope; Microscopic; Microscopy; neoplastic cell; next generation; novel; Optics; Phase; Physiological; prevent; Process; protein folding; prototype; Research; Research Personnel; Research Proposals; Resolution; responders and non-responders; response; Sampling; Scheme; Science; sensor; single cell analysis; Small Business Technology Transfer Research; spatiotemporal; Specimen; spectroscopic imaging; Spectrum Analysis; Speed; submicron; System; T-Lymphocyte; Techniques; Technology; Testing; Time; Tissues; tool; tumor; tumor progression; Universities; Validation; vibration; Visible Radiation; Water; Work