Super modulyo

The scaffold was washed with PBS and fixed with 2.5% glutaraldehyde at room temperature. After rinsing the scaffold with PBS and then with double deionized water (DDI), the fibrin scaffold was immersed in liquid nitrogen and kept in a freezer dryer (Super Modulyo, Edwards) [19 (link)]. The samples were coated with gold for 180 s by a sputter coater (SC7620, Emitech, UK) and observed under Scanning Electron Microscope (SEM, AIS2100, Seron technology, South Korea) at an accelerating voltage of 20 kV. The diameter of the fibers was calculated from SEM images by image analysis software (Image J, NIH, USA).

Frozen crickets were blanched for 2 min, chilled, and then quickly frozen to −40°C in a shock freezer. After that, the product was loaded into the freeze drier (Lyofast S08 and Super Modulyo, Edwards) set to −20°C, a vacuum was applied (±1–50 mbar absolute), and the temperature was slowly increased to +27°C. The condenser temperature was −60°C. The drying process lasted for ∼7 d. The weight of freeze-dried crickets was 24% of the initial fresh weight. The freeze-dried crickets were packaged in vacuum-sealed plastic bags and stored at room temperature. A few days before administering study meals, freeze-dried crickets were ground into a coarse cricket powder with a kitchen grinder. The cricket powder was stored in the freezer (−18°C) until meal preparation and administration.

Forty mL volumes of UV-B exposed (n = 3) and PAR only cultures (n = 3) were harvested at each time point by centrifugation at 4400 rpm for 20 min to produce a pellet and supernatant. The supernatants (40 mL) were collected and freeze-dried (Edwards, super modulyo) for 72 h. The remaining pellets were transferred into pre-weighed Eppendorf’s and freeze-dried for 24 h (Scanvac, CoolSafe^TM, LaboGene^TM, Vassingerød, Denmark) for dry weight measurements. Both pellets (PAR + UV-B and PAR only) and dried supernatant (PAR + UV-B only) were stored at −20 °C until analysis. OD was monitored using absorbance at 750 nm using a UV-visible spectrophotometer (Shimadzu, UV-2550, Kyoto, Japan).