Perfection v700

After ex vivo CMR, the heart was flash-frozen using liquid nitrogen in a 4.5 L cryogenic container (Fisher Scientific, Waltham, MA). The heart was sectioned into uniformly thick short axis slices using a commercial-grade prosciutto slicer (330 M; Berkel) obtaining slices of uniform thickness of approximately 2 mm to be matched to CMR studies. Slices were submerged in a PBS solution with 0.1 M triphenyl tetrazolium chloride (TTC) and incubated at 50 ° C for 15 min [18 (link)]. Slices were removed from TTC solution and mounted on glass slides with aqueous mounting media (Aquatex; EMD Millipore). TTC stained viable tissue deep red, distinguishing scar tissue and viable myocardium. Slides were imaged at 800 dpi resolution using an optical scanner (Perfection V700; Epson). Tissue sections from remote and infarct regions were selected for further histological analysis and fixed in 10% neutral buffered formalin. Tissue samples were stained with Masson’s trichrome. Collagen was stained blue whereas keratin, muscle fibers, and cytoplasm were stained red and nuclei dark red.

For root and hypocotyl length measurements, seedlings were grown on vertically oriented plates in the light (root) or dark (hypocotyl). Plates were scanned with an Epson Perfection V700 scanner and the length was measured using ImageJ 1.41 software (http://rsb.info.nih.gov/ij/).

Twitching motility was assessed on agar plates as described (16 (link)) (SI Appendix). Plates were photographed using an Epson Perfection V700 photo scanner.

Three tomato plants were randomly selected from each plot to dig a soil volume with a depth of about 0.4 m and a diameter of 0.2 m centered on the plant at 76 and 78 days after planting spring tomato and autumn tomato, respectively. The samples were placed in a 150 mesh sieve to rinse the roots. The roots were scanned by Epson Perfection V700 scanner to obtain the TIF diagram. Finally, the TIF diagram was processed by WinRHIZO Pro software to obtain the total root length, total number of root tips and bifurcation number of greenhouse tomatoes. The root activity of tomato was determined by triphenyltetrazolium chloride method (Li et al., 2020 (link)).

Root system characteristics were scanned by an Epson Perfection V700 scanner (Seiko Epson, Nagano, Japan) to obtain WinRHIZO PRO STD4800 (Regent, Quebec, MTL, Canada) root images. The processing software analyzed the root length, surface area, average root diameter, root tip, branch, and crossing number.
The maize roots were soaked in ethylenediaminetetraacetic acid solution (EDTA, Damao Chemical Reagent Co., Ltd., Tianjin, China) for 3 h, then thoroughly washed with deionized water. The tissue was dried at 75 °C for 72 h to obtain the shoot and root biomass.

Col-0 and randomly drawn (negative control) T-DNA lines in Table S7 were grown and phenotyped for GSA. Confirmed homozygous lines were obtained using PCR with the standard primers for the particular T-DNA and transgenic line (Table S8). For each line tested, two plates of seedlings were grown. Each plate had 16 T-DNA or transgenic seeds and 16 Col-0 seeds on opposite sides of the plate with four rows of four seeds each, and were grown in the same growth chamber for seven days, then scanned on an Epson Perfection V700 photo flatbed scanner into 24-bit RGB TIF image files at 600 dpi.

Upon termination of this experiment, cuttings were harvested in succession over three days, one treatment group per day. The order of harvesting was reversed between repetitions. For each cutting, roots, and shoots were separated with scissors, and shoot lengths were measured with a ruler. Roots were photographed using an Epson Perfection V700 scanner (Nagano, Japan). Root scan images were analyzed using WinRhizo^TM (Regent Instruments Inc., Quebéc City, QC, Canada) software, which provides biometric data outputs based on images. The parameters of root length and root diameter were obtained using WinRhizo^TM. The roots and shoots of each cutting were then dried at 70 °C for 72 h. Dry weights of roots and shoots were recorded on a precision scale. In repetition 2, the lengths of cuttings were also measured before the immersion step, and the difference in shoot lengths provided a measure of shoot growth during this experiment. Lastly, the dry weights of roots and shoots were divided to obtain a root/shoot dry weight ratio. In summary, the following seven biometric parameters were recorded: shoot length; shoot dry weight; root/shoot dry weight ratio; root length; root dry weight; root diameter; and shoot growth.