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Biological imaging with time domain fluorescence

Anand Kumar, PhD

Molecular imaging combines the use of disease targeted contrast agents with advanced imaging techniques to visualize disease processes in whole living organisms from the small animal to the human scale. Established techniques for whole animal imaging, including positron emission tomography (PET) and magnetic resonance imaging (MRI) offer high resolution and depth penetration but are expensive, highly complex in design and require significant expertise for their operation and maintenance. These techniques also offer limited flexibility for multiplexing, which refers to the ability to simultaneously visualize or quantify multiple biological components or pathways from the same subject. Optical techniques are emerging as a cost effective and portable alternative for functional contrast, and are particularly attractive given the spectral and fluorescence lifetime tunability of near infrared fluorophores. This allows the exciting possibility of multiplexing using fluorescence spectral and lifetime contrast.

My laboratory is focused on the development and application of whole-body time domain imaging techniques, with emphasis on exploiting lifetime contrast for functional imaging. Although fluorescence lifetime imaging has been widely used in microscopy using fluorescence lifetime imaging (FLIM), the application of lifetime imaging for macroscopic subjects has been limited by several challenges. This presentation will outline these challenges and present novel theoretical and experimental methods for tomographic lifetime imaging in the whole body. In particular, I will discuss a novel algorithm for tomographic lifetime multiplexing which allows the complete separation and 3-D localization of multiple lifetimes present simultaneously within biological tissue. Recent extensions of this work to the spatial frequency domain using modulated sources will also be discussed. I will then present in vivo applications of the technology pertaining to cancer and cardiac disease models. I will finally discuss recent progress towards clinical applications of fluorescence lifetime imaging using exogenous probes.

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