title: Imaging Cell-Matrix Interactions in 3D Collagen Hydrogel Culture Systems.
abstract: 3D hydrogels better replicate in vivo conditions, and yield different results from 2D substrates. However, imaging interactions between cells and the hydrogel microenvironment is challenging because of light diffraction and poor focal depth. Here, cryosectioning and vibrating microtomy methods and fixation protocols are compared. Collagen I/III hydrogel sections (20-100 µm) are fixed with paraformaldehyde (2%-4%) and structurally evaluated. Cryosectioning damaged hydrogels, and vibrating microtomy (100 µm, 2%) yielded the best preservation of microstructure and cell integrity. These results demonstrate a potential processing method that preserves hydrogel and cell integrity, permitting imaging of cell interactions with the microenvironment.
authors.affiliation: Department of Biomedical Engineering, College of Engineering, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Aaron R
authors.initials: AR
authors.lastname: Short
authors.affiliation: Department of Pathology and the Neurological Research Institute, College of Medicine, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Czeisler
authors.affiliation: Department of Biomedical Engineering, College of Engineering, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Stocker
authors.affiliation: Campus Microscopy and Imaging Facility, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Sara
authors.initials: S
authors.lastname: Cole
authors.affiliation: Department of Pathology and the Neurological Research Institute, College of Medicine, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: José Javier
authors.initials: JJ
authors.lastname: Otero
authors.affiliation: Department of Biomedical Engineering, College of Engineering, The Ohio State University, Columbus, OH, 43210, USA.|William G. Lowrie Department of Chemical and Biomolecular Engineering, College of Engineering, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Jessica O
authors.initials: JO
authors.lastname: Winter
mesh_terms: D002450:Cell Communication / Q000187:drug effects; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D060833:Cellular Microenvironment / Q000187:drug effects; D024042:Collagen Type I / Q000737:chemistry* / Q000627:therapeutic use; D005109:Extracellular Matrix / Q000187:drug effects*; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry* / Q000627:therapeutic use; D011108:Polymers / Q000737:chemistry
chemical_list: D024042:Collagen Type I; D011108:Polymers; D005557:Formaldehyde; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C003043:paraform
keywords: hydrogels; imaging; immunocytochemistry; sectioning; vibratomy