Recent advancements in mid-infrared (MIR) spectroscopy and nanofabrication have led to the development of Metasurface-Enhanced Infrared Reflection Spectroscopy (MEIRS), a field that explores the joint design of plasmonic metasurfaces and IR spectroscopy for biomedical research. The control over such high-dimensional data is unprecedented: chemical composition, protein structures, and cellular dynamics can be interactively monitored. However, traditional IR spectroscopy sensors lack the ability to capture critical morphology data. An “ultimate” IR imaging technique would enable real-time, label-free monitoring and imaging of live cells under diverse conditions. Chapter 1 sets the stage by providing the background and motivation for the research, highlighting the limitations of conventional techniques. The chapter delves into the theory of metasurfaces and their application in live cell spectroscopic sensing. Building on advancements in conventional optical microscopy, I propose metasurface-enhanced IR imaging as a transformative approach. Chapter 2 demonstrates the utility of MEIRS in tracking the effects of TRIP, a novel metal-based chemotherapeutic drug, on live A431 cells. This technique effectively observed apoptosis and loss of cell viability by monitoring changes in protein and lipid IR vibrations, as well as local refractive index shifts. The bio-orthogonal C≡O stretching signatures of TRIP provided precise drug delivery timelines, enhancing the interpretation of biological signaling. Chapter 3 documents the implementation and benchmarks of the inverted MIR laser stage-scanning microscope system. I started with a prototype and improved it through changes to hardware, optics, and software. The improved system achieved a spatial resolution of approximately 5 µm and a scan speed 55 times that of the prototype. I also demonstrated the imaging principles of metasurface-enhanced MIR chemical imaging with a PMMA nano-block experiment. Chapter 4 focuses on metasurface-enhanced MIR chemical imaging, providing non-destructive, label-free imaging of live 3T3-L1 cells. This technique offered exceptional vibrational contrast for cellular organelles. The method ensured high surface sensitivity, crucial for imaging cell morphology and adhesion. Chapter 5 introduces the advanced application of MIR chemical imaging on live 3T3-L1 cells, preserving cellular physiology while providing high-content imaging of cellular morphology and biochemistry. This method effectively profiled cellular responses to treatments, demonstrating significant changes in cell morphology and absorbance for nocodazole- and cytochalasin D-treated cells. In summary, this dissertation establishes metasurface-enhanced MIR imaging as a powerful tool for real-time, label-free imaging of live cells. These techniques provide valuable insights into cellular responses to treatments, enabling precise drug delivery monitoring, high-throughput screening, and detailed cell profiling. The advancements pave the way for further research and development in IR microscopy, with significant potential for clinical applications.