Molecules are the fundamental building blocks of all living matter. Disease is intrinsically linked to molecular malfunction: if undetected, tiny molecular errors can kill an organism. Visualising biology at the molecular level is thus key to advancing diagnostics and treatment. Yet, most current diagnostic imaging techniques only provide indirect molecular information that rarely allows drawing quantitatively meaningful conclusions. The PIRO project aims to overcome these limitations by developing a new vibrationally resolved holographic imaging methodology to enable high-resolution, quantitative, molecular imaging.
Compared to existing approaches, such as photothermal microscopy, a key aspect of PIRO is to shift from so-called “steady-state” heat-observations towards phototransient imaging. This paradigm shift conveniently eliminates heat-diffusion and time-dependent signal scaling effects thus ensuring high-quality, quantitative, vibrational imaging. Combined with phase-sensitive, holographic, widefield read-out the PIRO platform aims at providing truly quantitative, molecularly resolved, imaging at ~200 nm resolution across large fields of view at high throughput.
Technologically, PIRO’s ambitious goals are enabled by several core deliverables:
1. Advanced ultrafast infrared sources tailored for phototransient vibrational imaging.
2. A dedicated phototransient imaging system, the PIROscope, for rapid, high-resolution, label-free infrared imaging.
3. All-optical holographic lock-in schemes for widefield pump-probe imaging at the absolute sensitivity limit.
Enabled by these advances the PIRO project will then explore the fundamentally exciting transition from the transient to the well-studied steady state regime to both provide crucially necessary insight for informed imaging-system design and to uncover novel imaging methodologies based on dynamic observations. Finally, PIRO will take first steps towards enabling vibrationally-resolved phototransient diagnostics focusing on pressing challenges in the context of breast cancer diagnostics and antimicrobial resistance.
