CAREER: Unlocking Deep-Tissue Dynamics with Ultrabroadband Multiplex Label-Free Microscopy

Massachusetts Institute Of Technology, Cambridge MA

Investigators

Abstract

Microscopy has revolutionized our understanding of the biological world since its inception centuries ago. Label-free microscopy stands out for its ability to investigate living systems by capitalizing on the inherent optical signatures of cells. This eliminates the need for staining agents, thereby preserving the natural state of biological specimens and enabling an authentic representation of dynamic cellular processes. However, the reach of current label-free techniques is hindered by a fundamental compromise between molecular sensitivity, specificity, and imaging depth. The objective of this CAREER proposal is to overcome these barriers through the development of innovative light sources, excitation methods, and algorithmic reconstruction techniques. This project will develop new fiber optic sources and imaging platforms tailored for intrinsic contrast extraction from deep within living tissues, mitigating perturbation and enhancing molecular contrast. Such advancements promise to enable breakthroughs across physics, biology, and biomedical engineering, with direct implications for the real-time exploration and diagnosis of diseases in their native environments. In addition, this project will leverage existing outreach programs at MIT to provide K-12 students and teachers with access to the micro-world and understanding of the working principles of imaging by developing LEGO-based microscope modules. The education and outreach program are designed to spark curiosity and deepen STEM literacy among students from diverse backgrounds, with particular emphasis on engaging groups traditionally underrepresented in the sciences. By connecting research with education, this project aims to not only expand the frontiers of label-free imaging but also to cultivate a more inclusive and well-rounded future scientific community. The goal of this CAREER proposal is to establish a research program focusing on advanced label-free imaging technologies, with the potential to enhance disease diagnosis and therapeutic assessment. Current gold-standard methods for evaluating cellular metabolism and tissue changes, including histochemistry, fluorescent antibody tags, and genetically encoded imaging probes, are fundamentally limited by their necessity to physically, chemically, or genetically alter the biosystems. Label-free nonlinear microscopy circumvents these limitations by harnessing intrinsic molecular contrast within the tissue, offering real-time metabolic and structural imaging of living tissue without modifications. However, the reach of current label-free techniques is hindered by a fundamental compromise between molecular sensitivity, specificity, and imaging depth. This project seeks to enable transformative capabilities in label-free imaging through innovations in visible-to-SWIR (short-wavelength infrared) light generation and shaping, with three directions: (1) pioneer a high-peak-power, tunable, visible-to-SWIR fiber source for deep-tissue label-free multiplex imaging; (2) leverage ultrabroadband spectral shaping solutions; (3) optimize excitation and reconstruction for high-throughput hyperspectral label-free imaging. Achieving these imaging capabilities will represent a major milestone in our long-term effort of delivering label-free imaging technologies that rival or surpass the capabilities of label-based imaging, thereby advancing our characterization and understanding of dynamic biological processes with minimal perturbation and immediate clinical translation potential. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.