Biomedical imaging based on non-linear optical microscopy – deciphering the origin of intrinsic cellular autofluorescence targeting tissue development

Non-linear optical laser scanning microscopy has become an important imaging tool in the life-sciences. Operating in near infrared (NIR), it enables high resolution non-invasive 3D visualization of biological tissues.

A promising direction is the combination of multiphoton laser scanning microscopy with fluorescence life-time imaging (MPM-FLIM), as it can enable information about metabolic status.

However, development is hampered by incomplete understanding of the underlying molecular principles of intrinsic fluorescence.

Of particular importance is the contribution of nicotinamide adenine dinucleotide (NADH), a central coenzyme, and keratin, a highly abundant structural protein.

This project will tackle the challenge by posing two questions: (i) Is fluorescence lifetime of NADH influenced by protein binding or varying environmental conditions? (ii) What are the underlying molecular principles behind the intrinsic fluorescence of keratin?

The third task (iii) is to establish MPM combined with phasor-FLIM to enable studies of tissue status and development in an on-stage in vitro tissue culturing system.

The work builds on previous studies and techniques established in my lab, taking a novel direction by bringing in fundamental biochemistry and structural biology, in combination with new competence in the field of FLIM analysis.

The outcome is expected to promote translation of technology towards skin regeneration, pharmaceutical studies, and organ-on-a-chip models.