Ivis lumina 2 imaging system software

Harvested mango fruit with and without stem-end was used to measure the ROS level by fluorescence imaging, while using in-vivo Imaging System (IVIS Lumina II imaging system, PerkinElmer, USA). The ROS level was measured in mango stem-ends by staining with 10 µM of 2,7-dichlorodihydro fluorescein diacetate (H₂DCF-DA, Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBSX1), and incubation for 15 min. in the dark, then the stem-ends were washed twice with PBSX1 before IVIS evaluation. The stained fruit was exposed to 488 nm excitation and 510 nm emission wavelengths. The relative intensity of the fluorescent signal was calculated using IVIS Lumina II imaging system software (PerkinElmer, USA), as the average green fluorescence in three biological repeats for each treatment with/without stem-end.

Method for determining reactive oxygen species (ROS) was based on Galsurker et al. (2020) with slight modification [19 (link)]. Five ulvan-treated grapes and 5 DDW-treated grapes were tested on 2 and 4 DPT. The grapes were submerged in 10 μM DCF (2′,7′-Dichlorofluorescein, Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBS) and placed on an orbital shaker at 1.6 Hz for 15 min in the dark. Followed by de-staining with PBS solution on the orbital shaker for 1 min, and the washing repeated twice.
The fluorescence was measured at 488 nm excitation and 510 nm emission using in- vivo Imaging System (IVIS^®, PerkinElmer, Inc., Waltham, MA, USA) Results were calculated using IVIS^® Lumina II imaging system software (PerkinElmer, Inc., Waltham, MA, USA) and expressed in units of radiance [p/s/cm²/sr].