DOI: 10.5061/dryad.hx3ffbgn2
title: Data from: Preparation of isolated guard cells, containing cell walls, from Vicia faba

description: Stomatal movement, initiated by specialized epidermal cells known as guard
 cells (GCs), plays a pivotal role in plant gas exchange and water use
 efficiency. Despite protocols existing for isolating GCs through
 proplasting for carrying out biochemical, physiological, and molecular
 studies, protocols for isolating GCs with their cell walls still intact
 have been lacking in the literature. In this paper, we introduce a method
 for the isolation of complete GCs from Vicia faba and show their membrane
 to remain impermeable through propidium iodide staining. This methodology
 enables further in-depth analyses into the cell wall composition of GCs,
 facilitating our understanding of structure-function relationship
 governing reversible actuation within cells.
description: iGC yields were determined by calculating the dry weight
 percentages of remaining plant cells after isolation. Measurements were
 taken on the same day that leaves were collected and GCs were isolated.
 First, the harvested leaves were sorted by size (small, medium, and large)
 into approximately three equal piles. In order to document the size of
 leaves used to isolate GCs, leaf surface areas were measured using pixel
 counts in ImageJ (S1 Table). However, surface area measurements were not
 used in calculating 109<br>the final yield of iGCs.
 To calculate the final percentage of iGCs, dry weights from three
 different isolation experiments for the initial leaves and final iGCs were
 used. Dry weight of the leaves was determined by first measuring the wet
 weight of the three piles of leaves with a SECURA324-1S analytical
 balance. Their moisture content was then measured with a Mettler Toldeo
 Halogen Moisture Analyzer HC103 (115V). A standard drying program was run
 at 60 °C with 5 (1 mg/140 s) switch-off criterion. Using the wet weight
 and moisture content of each pile, the dry weight was calculated using
 Equation 1. Similar to the dry weight of the leaves, the dry weight of the
 iGCs was determined by first measuring the wet weight and then the
 moisture content. Mass dried (g) = Mass original −
 ((Moisture level x Mass original)/100)
description: # S1 Table Raw Data - Vicia faba dry weight & leaf size
 [https://doi.org/10.5061/dryad.hx3ffbgn2](https://doi.org/10.5061/dryad.hx3ffbgn2) Raw data for the S1 Table titled, "*Vicia faba* dry weight & leaf size." ## Description of the data and file structure Included are figures from which surface areas were calculated using ImageJ pixel counts. The raw data from each of these pixel counts can be found as tabs within the excel file titled, "S1 table raw," with the title structure "IMG_####-size#." The size (S, M, or L) within each tab indicates the leaf size of small, medium, or large. Also included within the excel file is a "Summary" tab, which contains raw data for the dry weight and leaf size of *Vicia faba* leaves that were used in this study. This information is subdivided into leaf size and then further subdivided into the number of leaves used, wet weight (g), dry weight (g), dry weight/leaf (g), Img file, and the leaf surface areas (cm^2). Also included is the percent moisture content (%MC) for each batch of leaves. The "Img file" corresponds to filenames of each image used for determining the average leaf surface area. Additional tabs included within the excel file contain raw data from each image.