Abstract
BackgroundThe in vivo observation of diffusible components, such as ions and small phenolic compounds, remains a challenge in turgid plant organs. The analytical techniques used to localize such components in water-rich tissue with a large field of view are lacking. It remains an issue to limit compound diffusion during sample preparation and observation processes.ResultsAn experimental setup involving the infusion staining of plant tissue and the cryo-fixation and cryo-sectioning of tissue samples followed by fluorescence cryo-observation by laser scanning confocal microscopy (LSCM) was developed. This setup was successfully applied to investigate the structure of the apple fruit cortex and table grape berry and was shown to be relevant for localizing calcium, potassium and flavonoid compounds.ConclusionThe cryo-approach was well adapted and opens new opportunities for imaging other diffusible components in hydrated tissues.
Highlights
The in vivo observation of diffusible components, such as ions and small phenolic compounds, remains a challenge in turgid plant organs
Staining approaches Staining of the sample with aqueous dye solutions must be completed prior to cryo-fixation. It was achieved by the infusion or perfusion (Fig. 1) of fresh samples using acridine orange (AO), a fluorescent dye for the cell walls and anionic sites of cell organelles, DNA and RNA [14,15,16]
The different cryo-fixation methods of AO infused samples were compared with regard to the apparent integrity of cell walls (Fig. 2)
Summary
The in vivo observation of diffusible components, such as ions and small phenolic compounds, remains a challenge in turgid plant organs. The analytical techniques used to localize such components in water-rich tissue with a large field of view are lacking. Plant growth involves intricate relations between cell water compartmentalization and cell wall mechanical properties [1] These relations involve cations for osmotic regulation and cell wall polysaccharide interactions, remodelling or deconstruction [2,3,4,5,6,7], but detailed knowledge on cation roles and interactions is impeded by their high mobility and/or low abundance in turgid tissue. Due to the high water content of growing plant tissue, restraining the diffusion of mobile ions and preserving tissue integrity remain a challenge [8,9,10]. Cryo-techniques coupled to cryo-observation in large fields of view using fluorescent techniques are suited to localize metallic cations and diffusible
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
