SBIR-STTR Award

Label-free sub-micron chemical imaging and spectroscopy has long been sought for visualization of biomolecules and materials in complex living systems- As stated in the DOE’s Biological and Environmental Research Program Office website, Bioimaging and measurement technologies that can resolve multiple key metabolic processes over time within or among cells will act as a crucial bridge toward linking molecular-scale information to whole-cell, systems-level understanding- Although Raman spectroscopy has made some inroads here, it is generally accepted by most Raman practitioners that the two major problems faced by Raman for cellular research, namely fluorescence and slow speed (tied to lack of sensitivity) limit its utility- IR micro-spectroscopy is a powerful technique but its use in cellular analysis in particular has been limited due to the following 3 key limitations: a) Spatial resolution limited by diffraction to ~ 10 µm- b) Inability for in-vivo imaging due to strong IR absorption by water, and c) accurate measurement of absorption in biological samples is hard to achieve because of scattering artifacts tied to sample heterogeneity, where the wavelength dependence of light scattering causes significant baseline artifacts- This proposal eliminates all 3 of the above limitations of IR micro spectroscopy while ensuring that the instrument is able to do the measurement simultaneously over a wide area (128*128 pixels)- The proposal is based on a recently published and patent-pending photo-thermal IR spectroscopy (PT-IR) breakthrough- Anasys previously pioneered the field of AFM (Atomic force microscope) based nanoscale photothermal IR Spectroscopy where an AFM probe detects the photothermal signal induced by IR absorption, thus obtaining ~ 10 nm spatial resolution [5]- Over the last several years, this product has been commercialized successfully in multiple industries valuing nanoscale- However, the constraints imposed by the use of an AFM, have restricted its adoption in cellular research in particular and life sciences in general- However, his experience of pioneering nanoscale photothermal IR with an AFM probe has given Anasys almost 10 years (including 3 years of R&D prior to the commercialization) of product and technology development experience in optimizing a signal based on photothermal physics, which will prove very useful for this project-

A key goal of bioenergy research is largely focused on converting low-energy, photosynthetically-generated plant sugars to high-value, high-energy bio-chemicals and biofuel products. Biofuel precursors are generated via cellular processes that occur on the microscopic scale, but currently available analytical instrumentation does not provide the ability to adequately probe critical biochemical processes on these length scales. Infrared spectroscopy, for example, arguably the most widely used technique for chemical analysis, has a fundamental spatial resolution limit on the scale of many microns. This phase II project aims to develop a novel analytical instrument based on Optical Photothermal Infrared (OPTIR) spectroscopy. The OPTIR technique can achieve ~10X better spatial resolution than conventional infrared spectroscopy (down to 500 nm). Phase I research demonstrated the feasibility of achieving >150X improvement in measurement sensitivity, achieving signal to noise values that are competitive with conventional infrared spectroscopy, but with much higher spatial resolution. Phase I research also demonstrated the ability to perform spectroscopy and chemical imaging on live biological cells, including mapping the distribution of lipids within the cells. The Phase II project is designed to mature technologies demonstrated in Phase I and extend these capabilities to achieve further improvements in measurement sensitivity, speed, and ease of use, as well as the ability to perform time-resolved spectroscopy and chemical imaging under physiological environments. To achieve further improvements in sensitivity and speed, Phase II activities will include additional technology development activities at PSC’s facilities in Santa Barbara CA, as well as at MIT Lincoln Labs. Additional engineering and applications efforts will be focused on automating alignment, optimization, measurement, and analysis steps, thus dramatically reducing the required operator skill level. This project will also focused on demonstrating the application of OPTIR to studies of functional metabolism for bioenergy applications. Towards that goal, the project includes close collaboration with bioenergy expert Prof. Basil Nikolau of Iowa State University and researchers at the Advanced Light Source at DOE’s Lawrence Berkeley Labs. Completion of this project will lead to the development of a new commercial instrument based on the OPTIR technology that will provide a robust ability to perform non-destructive chemical analysis for a key problems in bioenergy research, but also diverse applications in the life science and materials sciences.