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Rapid permeation of sucrose into rice callus cells as a component of cryoprotectant solutions

Ochoa-Castillo, Albert1; Volk, Gayle M.1; Levinger, Nancy E.1

Published Apr 15, 2026 on Dryad. https://doi.org/10.5061/dryad.6hdr7srfq

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Abstract

Sucrose was first included in plant vitrification solutions (PVS) in 1989 and has since remained a primary component of most formulations. Although sucrose is known to stabilize cell membranes, prevent protein denaturation, and promote cellular dehydration, its specific role in successful plant cryopreservation remains unclear. The study reported here evaluates sucrose permeation into rice callus cells for each step of a model cryopreservation cycle. By deuterating sucrose, its C–D bond can be tracked using coherent anti‑Stokes Raman scattering microscopy, a highly sensitive, non‑invasive vibrational microscopy technique, thus converting the molecule into its own imaging agent. Results show that sucrose permeates cells quickly, within 6 s of initial exposure, and takes only 20‑30 s to reach steady‑state concentration. Sucrose also permeates the cells regardless of pre‑existing intracellular sucrose from prior exposure in the model cryopreservation cycle. Beyond the permeation of sucrose as a cryoprotecting agent, the results presented here suggest that this method could measure permeation by sucrose into biological tissue or biomaterials, and can be extended to other molecules, such as the permeation of herbicides or drugs, showing kinetics and sequestration into live cells and tissue.