Phyto_cone survival Fig 1
Description
800 different extracts were tested in HCS for chicken cone photoreceptor survival from a tropically diverse range of plants at concentrations of 10 and 100 µg/ml . The full cone survival data inferred from visinin immunostaining across two hundred 96-well plates is presented.
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Plant materials were extracted using an ASE 200 system (Dionex) under nitrogen (6–8 bar). Four extract types were prepared: methanolic polyamide (MP), ethyl acetate (EA), depleted ethyl acetate (DEA), and depleted methanol (DM). For MP extraction, 5–6 g of powdered plant material was mixed with 5 g of polyamide (SC6, Macherey-Nagel), 10 g of plant material was used for EA, DEA, and DM extractions. DEA and DM preparations included a preliminary heptane depletion step, followed by extraction with ethyl acetate or methanol, respectively. Extractions were performed at 60°C using four 3-min cycles for MP and five 3-min cycles for EA, DEA, and DM, followed by a 60% solvent flush and a 120-s nitrogen purge. Extracts were collected, concentrated at 40°C under reduced pressure using a Büchi rotary evaporator, reconstituted in water, and lyophilized for 48 h. A library comprising 800 extracts from 200 plant species was generated. An aliquot of 2 mg of each dried extract was transferred into individual 4-mL glass vials, uniquely coded, and stored at −80°C. Extraction yield was determined gravimetrically as the ratio of dried extract mass to initial plant material mass × 100. The selection criterion was the viability factor: the average cone survival across 4 replicate wells divided by the average survival across 19 negative control wells. A viability factor ≧2 was chosen arbitrarily as the cut-off. Ultimately, two hundred 96-well plates requiring 700 fertilized chicken eggs were used to produce the full data.
Institutions
- Institut de la VisionÎle-de-France, Paris