A spectrochemical imaging system for rapid and nondisruptive screening for pluripotency and differentiation in cultures of embryonic stem cells

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Embryonic stem cells (ESC) offer tremendous potential for addressing a host of important biomedical problems, both in clinical and laboratory settings. This potential has generated extraordinary excitement over the use of these cells to devise novel cellular therapies for many currently debilitating and costly human diseases (e.g., diabetes and Parkinson's disease), as well as irreparable injuries (e.g., spinal cord paralysis). There are, however, many hurdles to overcome between our present state and new therapeutic applications. One of the fundamental technological obstacles that dramatically slows progress in this area is the lack of rapid, nondestructive assays for distinguishing ESC from other (differentiated) cells within a growing culture. We are proposing to develop such a rapid, nondestructive, non-contact screening methodology based on an optical technique called coherent anti-Stokes Raman scattering (CARS) microscopy. This nonlinear optical method permits imaging of ESCs in situ within a culture and exploits the rich chemical information carried by the CARS signal to distinguish stem cells from differentiating cells. Specifically, we are proposing i) to develop a robust optical component system for efficient exposure and collection of CARS spectra and advanced signal-processing methods to aid the interpretation of CARS spectra of ESC; ii) to undertake the first spectrochemical imaging investigation involving human ESC to identify and characterize the spectral features most sensitive to cellular changes (and/or the rate of such changes) accompanying differentiation; iii) to prepare this technology for installation at an existing bioimaging core facility that provides access by stem cell biologists and other medical scientists. This project will lay the foundation for accelerated progress toward developing human embryonic stem cell-based therapies and will facilitate the broader use of these cells to identify new drugs that stimulate or repress particular regenerative events in vivo.