title: Comparing different methods to fix and to dehydrate cells on alginate hydrogel scaffolds using scanning electron microscopy.
abstract: Scanning electron microscopy (SEM) is commonly used in the analysis of scaffolds morphology, as well as cell attachment, morphology and spreading on to the scaffolds. However, so far a specific methodology to prepare the alginate hydrogel (AH) scaffolds for SEM analysis has not been evaluated. This study compared different methods to fix/dehydrate cells in AH scaffolds for SEM analysis. AH scaffolds were prepared and seeded with NIH/3T3 cell line; fixed with glutaraldehyde, osmium tetroxide, or the freeze drying method and analyzed by SEM. Results demonstrated that the freeze dried method interferes less with cell morphology and density, and preserves the scaffolds structure. The fixation with glutaraldehyde did not affect cells morphology and density; however, the scaffolds morphology was affected in some level. The fixation with osmium tetroxide interfered in the natural structure of cells and scaffold. In conclusion the freeze drying and glutaraldehyde are suitable methods for cell fixation in AH scaffold for SEM, although scaffolds structure seems to be affected by glutaraldehyde.
authors: Nucleus of Cellular and Tecidual Biology (NCTBio), Post-Graduate Program in Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil.
authors: Bianca Palma
authors: BP
authors: Santana
authors: Nucleus of Cellular and Tecidual Biology (NCTBio), Post-Graduate Program in Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil.|Post-Graduate Program in Health and Behavior, Catholic University of Pelotas, Pelotas, RS, Brazil.
authors: Fernanda
authors: F
authors: Nedel
authors: Nucleus of Cellular and Tecidual Biology (NCTBio), Post-Graduate Program in Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil.
authors: Camila
authors: C
authors: Perelló Ferrúa
authors: Technology Development Center, Federal University of Pelotas, Pelotas, RS, Brazil.
authors: Ricardo
authors: R
authors: Marques e Silva
authors: Nucleus of Cellular and Tecidual Biology (NCTBio), Post-Graduate Program in Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil.
authors: Adriana Fernandes
authors: AF
authors: da Silva
authors: Nucleus of Cellular and Tecidual Biology (NCTBio), Post-Graduate Program in Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil.
authors: Flávio Fernando
authors: FF
authors: Demarco
authors: Technology Development Center, Federal University of Pelotas, Pelotas, RS, Brazil.
authors: Neftali
authors: N
authors: Lenin Villarreal Carreño
mesh_terms: D000464:Alginates / Q000737:chemistry; D000818:Animals; D002477:Cells / Q000737:chemistry* / Q000648:ultrastructure; D005612:Freeze Drying / Q000379:methods*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D016707:Tissue Fixation / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid
keywords: SEM; freeze drying; glutaraldehyde; osmium tetroxide; tissue engineering