Labophot pol

Sample analysis was carried out following the protocol of histological assessments. Portions of approximately 6 mm³ of hydrogel scaffolds after 14, 22, 43, and 75 days of culture were fixed by immersion in 10% (v/v) buffered-saline formaldehyde pH 7.2–7.4 at 4 °C, dehydrated with alcohol, and embedded in paraffin. Then, the pieces were cut in ≈5 μm histological sections with a microtome (Micron, Germany) and mounted on slides.
Paraffin-embedded sections (5 μm thick) were deparaffinised through xylene and a graded alcohol series, and then stained for 30 minutes with Picrosirius Red (0.1% of Sirius Red in saturated aqueous picric acid), as described by Junqueira et al.^{36 (link)} for collagen bundle detection. Sections were then mounted for observation under polarized light microscopy (Nikon Labophot-Pol).

Reaction mixtures were examined under a Nikon LABOPHOT- POL (Melville, NY, USA) optical polarizing microscope. Crystals suitable for SCXRD were suspended in Paratone-N oil (Hampton Research, Alisa-Viejo, CA, USA) and mounted on nylon loops or glass fibers. Data were collected on a Bruker diffractometer equipped with a PLATFORM three-circle goniometer and an APEX II charge-coupled device area detector (Bruker AXS, Madison, WI, USA). For structure solution, a hemisphere of unique data was collected with Mo-Kα radiation (λ = 0.71054 Å) using strategies of scans about the ω and φ axes with 0.5° frame widths. Crystals were cooled to −100°C during collection under a cold stream of N₂ gas using an N-HeliX cryostat (Oxford Cryosystems, Oxford, UK).