Label-Free detection of antisense oligonucleotide uptake in Murine cells using hyperspectral CARS imaging

Prabuddha Mukherjee; Aneesh Alex; Edita Aksamitiene; Derek Shi; Darold R. Spillman Jr.; Marina Marjanovic; Michael Fazio; Punit Seth; Kendal Frazier; Steve R. Hood; Stephen A. Boppart

doi:10.1117/12.2578061

5 March 2021 Label-Free detection of antisense oligonucleotide uptake in Murine cells using hyperspectral CARS imaging

Prabuddha Mukherjee, Aneesh Alex, Edita Aksamitiene, Derek Shi, Darold R. Spillman Jr., Marina Marjanovic, Michael Fazio, Punit Seth, Kendal Frazier, Steve R. Hood, Stephen A. Boppart

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Proceedings Volume 11656, Advanced Chemical Microscopy for Life Science and Translational Medicine 2021; 116561F (2021) https://doi.org/10.1117/12.2578061
Event: SPIE BiOS, 2021, Online Only

Abstract

Antisense oligonucleotides (ASOs) are single stranded negatively charged molecules which downregulate the translation of specific target messenger RNA (mRNA). Chemically modified ASOs with phosphorothioate (PS) linkages have been extensively studied as research tools and as clinical therapeutics and nine oligonucleotide-based drugs have been approved by regulatory agencies. While several cell surface proteins that bind PS-ASOs and mediate their cellular uptake have been identified, the mechanisms leading to productive internalization of PS-ASOs are not well understood. We demonstrate the potential of hyperspectral CARS imaging to detect the intracellular presence of ASOs in a label-free manner.

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Prabuddha Mukherjee, Aneesh Alex, Edita Aksamitiene, Derek Shi, Darold R. Spillman Jr., Marina Marjanovic, Michael Fazio, Punit Seth, Kendal Frazier, Steve R. Hood, and Stephen A. Boppart "Label-Free detection of antisense oligonucleotide uptake in Murine cells using hyperspectral CARS imaging", Proc. SPIE 11656, Advanced Chemical Microscopy for Life Science and Translational Medicine 2021, 116561F (5 March 2021); https://doi.org/10.1117/12.2578061

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KEYWORDS

CARS tomography

Hyperspectral imaging

Microscopy

Imaging spectroscopy

In vivo imaging

Luminescence

Molecules

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