DOI: 10.5061/dryad.np5hqc02q
title: Data from: Direct evidence that cryoprotectant mixtures facilitate individual component permeation into living plant cells

description: The fundamental interactions between plant cells and cryoprotectants
 during vitrification are understudied in the field of plant
 cryopreservation. Within this area research, real time cryoprotectant
 permeation into plant cells is even less documented. In this study, we
 monitor the real time permeation of individual cryoprotectants into rice
 callus cells when in mixtures with other cryoprotectants. Specifically, we
 use coherent anti-Stokes Raman scattering (CARS) microscopy to observe the
 permeation of individually deuterated DMSO, ethylene glycol, and glycerol
 in plant vitrification solution 2 (PVS2) by probing vibrational
 frequencies that correspond to C-D stretching modes of the cryoprotectant
 molecules. Additionally, we measure cell plasma membrane responses to PVS2
 exposure using brightfield microscopy. We conclude that the permeation of
 PVS2 components into plant cells occurs faster than the first cell plasma
 membrane responses observed and therefore permeation and cell plasma
 membrane response do not appear to be directly correlated. In addition, we
 observe that cryoprotectant permeation into plant cells occurs more
 quickly and more uniformly when cryoprotectants are in PVS2 solution than
 when they are in single component aqueous solutions.
description: # Data from: Direct evidence that cryoprotectant mixtures facilitate
 individual component permeation into living plant cells
 [https://doi.org/10.5061/dryad.np5hqc02q](https://doi.org/10.5061/dryad.np5hqc02q) ## Description of the data and file structure This README file was generated on 2024-06-12 by Kylie Pearce. ## GENERAL INFORMATION Author/Principal Investigator Information Name: Nancy Levinger ORCID: 0000-0001-9624-4867 Institution: Colorado State University Email Address: [nancy.levinger@colostate.edu](mailto:nancy.levinger@colostate.edu) Author/Co-investigator Information Name: Kylie Pearce ORCID: 0000-0002-8629-5932 Institution: Colorado State University Email: [kylie.pearce@colostate.edu](mailto:kylie.pearce@colostate.edu) Date of data collection: 2022-02-16 to 2023-01-26 Geographic location of data collection: Fort Collins, CO, United States and Aroura, CO, United State Information about funding sources that supported the collection of the data: Main funding sources for this project include USDA NIFA Grant Number 12835363 and USDA ARS Non-Assistance Cooperative Agreement 58-3012-9-017. Author Fionna M. D. Samuels acknowledges support from the NSF Graduate Research Fellowship Program fellow number 2019273956. ## SHARING/ACCESS INFORMATION Licenses/restrictions placed on the data: none Links to publications that cite or use the data: [https://doi.org/10.1016/j.cryobiol.2024.104928](https://doi.org/10.1016/j.cryobiol.2024.104928) ## DATA & FILE OVERVIEW ### Folder List: Brightfield Experiments_Cell Response Times Statistical Analysis Brightfield Experiments_Raw Data and ROIs CARS Experiments_Analysis and Raw Data Relationship between folders: Data provided here is divided into two categories: brightfield microscopy data and CARS microscopy data. The folder “Brightfield Experiments_Cell Response Times Statistical Analysis” contains the analysis of the raw data contained in the folder “Brightfield Experiments_Raw Data and ROIs”. The folder “CARS Experiments_Analysis and Raw Data” contains the analysis and raw data for all coherent anti-stokes Raman scattering (CARS) microscopy experiments. METHODOLOGICAL INFORMATION Description of methods used for collection/generation of data: The two methods used for generating the data presented here are brightfield microscopy and coherent anti-stokes Raman scattering (CARS) microscopy. A full description of the experimental protocol is available in the full paper’s methods section. Methods for processing the data: All microscopy images present in the main text, supplementary information, and this dataset were analyzed using Fiji. First, the native file formats (.avi for brightfield data and .oib for CARS data) were converted to .tiff files for ease of use. Next, a Virdis color scheme was applied to CARS microscopy data. Finally, in both brightfield and CARS data, regions of interest (ROIs) were generated to identify individual callus cells and, for CARS data specifically, portions of the background. Any current version of Fiji can be used to analyze the raw data however, we recommend downloading the Stackreg plugin to make analysis of unstable images easier. Additional information about how the data processed with Fiji is further analyzed is provided in each folder’s own README file. The R code present in each file is compatible with version 2022.12.0. Environmental/experimental conditions: All experiments were performed at 25°C. People involved with sample collection, processing, analysis and/or submission: Kylie C. Pearce and Fionna M. D. Samuels