Presentation Details
| Quantification of fluorescence responses in a newly developed, dryable cell line expressing odorant receptors Andisheh Balouchi1, Redwan Haider1, Roy Anderson1, Richard Cornette2, Takahiro Kikawada2, Ricardo Araneda1, Elisabeth Smela1. 1University of Maryland, College Park, MD, USA.2NARO, Tsukuba, Japan |
Abstract
Olfactory sensors are powerful tools for potential applications in diverse fields from environmental monitoring to medical diagnostics. Specifically, cell-based sensors utilizing insect odorant receptors (ORs) show great potential capability to detect and distinguish odors. However, the limited durability of living cells outside aqueous culture conditions constrains the sensor’s storability and function. To overcome this issue, we utilized a novel desiccatable cell line Pv11, derived from Polypedilum vanderplanki, which can survive complete dehydration and storage in the dry state for over a year. The Pv11 cells were engineered to express a specific Drosophila melanogaster OR (Or47a) with its corresponding co-receptor Orco and the intracellular calcium indicator GCaMP6f. In our previous study, we demonstrated that Pv11-Or47a cells respond to their cognate ligand, pentyl acetate (PA), 12 hours after rehydration, in a dose-dependent manner and to repeated exposures. Here, we improve quantification of fluorescence responses in Pv11 cells by normalizing readouts to viable cells using Hoechst 33342 to stain the nuclei and ethidium homodimer-1 to identify membrane-compromised (dead) cells. We found that the cells’ response to PA 10 mM is undiminished even 12 hours after staining. These findings support the development of a viability-normalizing method for storable, cell-based olfactory sensors using engineered, desiccation-tolerant Pv11-Or47a cells.
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No part of this publication may be reproduced, distributed, or transmitted in any form or by any means, including photocopying, recording, or other electronic or mechanical methods, without the prior written permission of the author.
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