Homogenate was prepared by a suspension of dry S. platensis in deionised water (in a ratio 1:10) and mixing (Thermomix; Vorkwerk Ltd., Poland) at 37 °C for 40 min. (500 rpm). The obtained solution was centrifuged for 20 min. (4600 rpm) (Heraeus Megafuge 40, rotor TX-750, Thermo Scientific, Waltham, MA, USA). Supernatant was separated and treated as an algal filtrate (F)—100% and then (1) foliarly applied to the sprouts as an aqueous solution at different concentrations (5, 7, 10, 15, 20 and 25%, v/v). The 15% concentration was used for (2) seeds soaking for different time spans–1, 3, 6, 12, 18, 24, 36 and 48 h. The remaining solid residue, treated as an algal homogenate (H)–100%, was diluted with deionised water (1:1) and used for (3) seeds coating (doses–100, 300, 500, 700 µL per 1.5 g of radish seeds). This treatment was performed using vortex-type shaker for 10 min. The concentrations were selected on the basis of our previous studies (Michalak et al. 2017 (link)).
Heraeus megafuge 40
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Heraeus Megafuge 40 is a high-performance centrifuge designed for a variety of laboratory applications. It features a maximum speed of 14,000 rpm and a maximum RCF of 21,382 x g, making it suitable for a wide range of sample volumes and types. The Megafuge 40 is equipped with a user-friendly control panel and offers temperature control, selectable acceleration and deceleration rates, and various rotor options to accommodate different sample requirements.
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4 protocols using heraeus megafuge 40
1
Spirulina Extracts for Radish Seed Treatment
2
Quantification of Total Phenolic Compounds
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The total phenolic compounds (TPC) content (mg of gallic acid equivalents (GAE)·100 g−1 FW) was determined in accordance to the Folin-Ciocalteu method proposed by Jałoszyński et al. [87 ] with slight alterations [14 (link),15 (link)]. The fresh and fragmented (using Thermomix) shoots and roots (~2 g) were placed in 50 mL falcon tubes and the aqueous methanol (20 mL, 80%) was added. The test tubes were sonicated (Bandelin Sonorex RK 100 H, Berlin, Germany) for 15 min and centrifuged (10 min, 4500 rpm) (Heraeus Megafuge 40, rotor TX-750, Thermo Scientific). To the supernatants (0.1 mL), the Folin-Ciocalteu’s phenol reagent (0.2 mL) and distilled water (2 mL) were added and left to stand at room temperature in the dark for 3 min. Afterwards, sodium carbonate (1 mL, 20%) was added, and the reaction mixtures were kept in the dark for 1 h. The absorbance at 765 nm was measured (HACH DR1900 spectrophotometer, four replicates).
3
Ultrasound-Assisted Biomass Extraction
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Bioproducts were obtained by means of ultrasound-assisted extraction (UAE) using the homogeniser UP 50 H (Hielscher Ultrasonics GmbH, Teltow, Germany). Dried and ground (500 μm mesh size) biomass was well-mixed in a glass beaker with deionised water (ratio 1:20 w/v), left for maceration (30 min, room temperature), sonicated (30 min), and centrifuged (4500 rpm, 10 min, Heraeus Megafuge 40, rotor TX-750, Thermo Scientific, Waltham, MA, USA). The obtained supernatants were used for analyses.
4
Ultrastructural Insights of Plasmodium falciparum
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Pf3D7 parasites treated with 10 x EC50 of 3 or the same concentration of vehicle control (DMSO) were spun in Eppendorf tubes in a Thermo Scientific Heraeus Megafuge 40 centrifuge at 2000 rpm for 3 min to pellet the RBC. The resulting pellets were fixed in 2.5% glutaraldehyde in 0.1 M Sorensen’s phosphate buffer (pH 7.2, 300 mOsmol) at 4 °C. Taking care not to completely resuspend the RBC pellets, fragments of the pellets were intentionally produced by gently surging with analogous buffer delivered via pipette tip. Due to the fragility of the fragments, gentle inversion of the Eppendorf tubes was used to mix reagents during processing rather than agitation. After washing the fragments in buffer for 15 min, they were immersed in a secondary fixative of 1% osmium tetroxide in buffer for 60 min. The fragments were dehydrated in a graded ethanol series, infiltrated with absolute dry ethanol mixed with Spurr’s Resin for 30 min and then two changes of 100% Spurr’s Resin for 60 min. Singular fragments were embedded in resin in BEEM polyethylene capsules and polymerized overnight at 65 °C. Ultrathin sections were stained with uranyl acetate, followed by lead citrate. Sections were examined using a JEOL TEM – 1400 120kV TEM and images acquired with a Gatan UltraScan 1000 camera.
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