SBIR-STTR Award

Anasys Instruments proposes to develop an optical platform that combines confocal Raman spectroscopy with infrared spectroscopy with sub-micron spatial resolution on a single optical microscope based platform (patents pending). Infrared spectroscopy and Raman spectroscopy are separately extremely successful and complementary techniques for chemical composition analysis. Infrared spectroscopy characterizes samples by detecting absorption of infrared light by dipole moment changes induced in molecular bond vibrations. Raman spectroscopy detects scattered light that wavelength shifted by a sample due to molecular vibrations that induce changes in polarizability. The techniques are highly complementary because molecular vibrations that are strong IR absorbers are typically weak Raman scatterers and vice versa. Conventional IR spectroscopy has two significant limit fundamental limitations: (1) spatial resolution limited by optical diffraction from its longer wavelengths to the scale of many microns; (2) sample preparation is often complex, requiring preparation of thin IR transparent samples. These limitation, along with sample preparation complexities have limited applicability of IR spectroscopy in many life sciences applications. Differences in IR and Raman wavelength ranges, light sources and detectors have also made it impossible to obtain IR and Raman spectroscopy from the same sample region. As a result, there is no currently available instrument that can perform infrared and Raman analysis on the same sample with sub-micron spatial resolution for both techniques. We propose to overcome this limitation developing and demonstrating the applicability of the IRaman platform which will provide simultaneous and complementary IR and Raman analysis in the same instrument. This project will leverage the recent invention of sub-micron photothermal IR spectroscopy, a technique that uses a tightly focused visible laser to probe the infrared absorption on a scale ~10X smaller than the IR optical diffraction limit. Complementary Raman measurements will be obtained by using the same visible laser beam in combination with a Raman spectrometer to excite and detect Raman scattering in the sample. A Prototype instrument will be built to demonstrate utility of simultaneous IR/Raman technique in a couple of high value biomedical applications. Anasys has extensive experience in successful commercialization of breakthrough IR analysis products based on photothermal physics Anasys Instruments Proprietary and Confidential 1

Project Terms:
absorption; Address; Aging; base; bioimaging; Biological; Biological Sciences; Biomedical Research; Cells; Cellular biology; Chemicals; commercialization; Complex; Computer software; detector; dipole moment; Drug Evaluation; Erythrocytes; Excipients; experience; Goals; Heating; Hybrids; Image; infrared spectroscopy; instrument; invention; Lasers; Legal patent; Light; light scattering; Maps; Measurement; Measures; Microscope; Microscopic; Molecular; Optics; Oranges; particle; Performance; performance tests; Pharmaceutical Preparations; Pharmacologic Substance; Physics; Preparation; prototype; Raman Spectrum Analysis; Research; Resolution; response; Sampling; Science; Small Business Innovation Research Grant; Source; Specimen; spectroscopic imaging; submicron; Tablets; Techniques; Temperature; Thinness; vibration;

This Phase II proposal aims to develop and commercialize IR+Raman a breakthrough instrument that will for the first time enable simultaneous infrared (IR) and Raman spectroscopy on the same instrument with sub-micron spatial resolution. This project is well aligned with NIH goals as it incorporates several key thrusts of the National Institute of Biomedical Imaging and Bioengineering, including optical imaging and spectroscopy, infrared imaging, confocal microscopy, and multimodal imaging. IR and Raman have gained interest in investigating the composition and molecular structure of biological materials as they operate label free and are sensitive towards macromolecular composition, such as proteins, lipids, nucleic acids and carbohydrates, as well as the detection of isotopic labelling of these macromolecules and even smaller metabolites. Both Infrared and Raman spectroscopies are widely used in analytical laboratories and are often referred to as “complementary techniques” as they both probe different types of molecular vibrations. For example, IR spectroscopy is very sensitive to protein secondary structure, whereas Raman is particularly sensitive to lipids as well as certain amino acids. And in pharma applications Raman is more sensitive to drugs, whereas IR is more sensitive to excipients (additives) that often have weak Raman signals and/or have large fluorescent backgrounds. Raman can achieve sub-micron spatial resolution, but IR is limited by the longer excitation wavelengths to spatial resolution ~10 um. This project aims to overcome this limitation by providing IR and Raman spectroscopy, both at sub-micron spatial resolution. A compelling recent example of the power of the multimodal combination of IR and Raman in health sciences involved analysis of malaria parasite infected red blood cells (D. Perez-Guaita et al Vib. Spectrosc. 91, 46-58 (2017)). The research showed “that the combination of both techniques provides complementary information not evident” using the techniques individually. This research was performed however using a painstaking process of separately and sequentially measuring the exact same cell locations in two different instruments, requiring substantial additional time and cost. This proposal aims to develop an instrument that makes simultaneous IR and Raman measurements simple, robust, and routine. This project will leverage successful Phase I research to develop and commercialize a new optical microscope-based platform that can perform simultaneous IR and Raman on the same instrument. The project will involve a collaboration between proposer Photothermal Spectroscopy Corp and Dr. Ji-Xin Cheng (Boston University) and Dr. Lynne Taylor (Purdue). The team at photothermal will design and build a next generation IR+Raman instrument to overcome key limitations and expand the capabilities over the prototype developed in Phase I. The two year project will develop alpha and beta prototype units for applications testing at Photothermal’s applications lab in Santa Barbara, CA, and will install a beta unit in the labs of Prof. Lynne Taylor at Purdue University, with a focus on demonstrating applicability of IR+Raman to key problems in pharmaceutical sciences. Photothermal scientists will also collaborate closely with Prof. Cheng’s group at Boston University in cell biology, specifically related to investigate antibody susceptibility at the single bacterium level. The beta IR+Raman will also be used to investigate other applications in cells/tissue and microplastics characterization.

Public Health Relevance Statement:
Project narrative Raman spectroscopy and infrared spectroscopy are complementary analytical techniques that can provide critical information about chemical composition on the microscopic scale, but to date incompatibilities between the two techniques has made it impossible to perform simultaneous IR and Raman measurements, especially on the scale required for most biomedical samples. This SBIR project will overcome the previous barrier, developing a breakthrough instrument for biomedical imaging that will enable simultaneous infrared and Raman spectroscopy on the same optical microscope based platform while providing 10X better spatial resolution than conventional infrared spectroscopy. The new instrument will have profound effects on biomedical research, including dramatically improved characterization of pharmaceuticals and biological cells/tissue that will lead to improved drug efficacy and stability and provide fundamental insights into disease and human health.

Project Terms:
absorption; Address; Amino Acids; Antibiotic susceptibility; Antibiotics; Antibodies; Bacteria; base; Biocompatible Materials; bioimaging; Biological; Biological Sciences; Biology; Biomedical Research; Boston; Carbohydrates; Cells; Cellular biology; Chemicals; Collaborations; Confocal Microscopy; controlled release; cost; crystallinity; Crystallization; Data; design; Detection; detector; Disease; Drops; drug efficacy; Drug Formulations; Drug Stability; Engineering; Environmental Impact; Erythrocytes; Excipients; extracellular vesicles; Formulation; Goals; Health; Health Sciences; Heating; Hour; Human; Image; improved; Individual; infrared spectroscopy; insight; instrument; instrumentation; interest; Isotope Labeling; Label; Laboratories; Lasers; Lipids; Location; macromolecule; Malaria; Measurement; Measures; Mechanics; Metabolic; Microscope; Microscopic; microscopic imaging; Molecular; Molecular Structure; Multimodal Imaging; multimodality; National Institute of Biomedical Imaging and Bioengineering; next generation; Nucleic Acids; optical imaging; Optics; Parasites; Particle Size; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Predisposition; Process; Proteins; prototype; Raman Spectrum Analysis; Research; Resolution; Sampling; Scanning; Science; Scientist; Secondary Protein Structure; Signal Transduction; Small Business Innovation Research Grant; software development; Solubility; spectroscopic imaging; Spectrum Analysis; submicron; Techniques; temporal measurement; Testing; Time; Tissues; transmission process; United States National Institutes of Health; Universities; vibration