To evaluate the morphology of the Hy-Tissue Nanofat, SVF
whole-mount assay was performed, as reported in Busato et al.30 (link)
The emulsion was swiped in a histological glass and stained with
toluidine blue (Sigma-Aldrich). All slides were examined under an Olympus BX-51
microscope (Olympus, Tokyo, Japan) equipped with a digital camera (DKY-F58 CCD
JVC, Yokohama, Japan). In addition, for a deeper morphological understanding,
the Hy-Tissue Nanofat-SVF was studied in scanning electron
microscopy (SEM). The sample was fixed with glutaraldehyde 2% diluted in 0.1 M
phosphate buffer (pH 7.4) for 2 h at 4°C and post-fixed in 1% osmium tetroxide
(OsO4) diluted in 0.2 M potassium hexacyanoferrate for 1 h at
4°C. The samples were dehydrated in a graded concentration of ethanol, followed
by a critical point dryer (CPD 030, Balzers, Vaduz, Liechtenstein), mounted to
stubs with colloidal silver and sputtered with gold by an MED 010 coater
(Balzers), and examined with FEI XL30 scanning electron microscope (FEI Company,
Eindhoven, The Netherlands). All the morphological analyses were performed on
each sample collected from every patient.
whole-mount assay was performed, as reported in Busato et al.30 (link)
The emulsion was swiped in a histological glass and stained with
toluidine blue (Sigma-Aldrich). All slides were examined under an Olympus BX-51
microscope (Olympus, Tokyo, Japan) equipped with a digital camera (DKY-F58 CCD
JVC, Yokohama, Japan). In addition, for a deeper morphological understanding,
the Hy-Tissue Nanofat-SVF was studied in scanning electron
microscopy (SEM). The sample was fixed with glutaraldehyde 2% diluted in 0.1 M
phosphate buffer (pH 7.4) for 2 h at 4°C and post-fixed in 1% osmium tetroxide
(OsO4) diluted in 0.2 M potassium hexacyanoferrate for 1 h at
4°C. The samples were dehydrated in a graded concentration of ethanol, followed
by a critical point dryer (CPD 030, Balzers, Vaduz, Liechtenstein), mounted to
stubs with colloidal silver and sputtered with gold by an MED 010 coater
(Balzers), and examined with FEI XL30 scanning electron microscope (FEI Company,
Eindhoven, The Netherlands). All the morphological analyses were performed on
each sample collected from every patient.
