title: Cryo-immunoelectron microscopy of adherent cells improved by the use of electrospun cell culture substrates.
abstract: Electrospun nanofibres are an excellent cell culture substrate, enabling the fast and non-disruptive harvest and transfer of adherent cells for microscopical and biochemical analyses. Metabolic activity and cellular structures are maintained during the only half a minute-long harvest and transfer process. We show here that such samples can be optimally processed by means of cryofixation combined either with freeze-substitution, sample rehydration and cryosection-immunolabelling or with freeze-fracture replica-immunolabelling. Moreover, electrospun fibre substrates are equally suitable for complementary approaches, such as biochemistry, fluorescence microscopy and cytochemistry.
authors: Department of Therapeutic Radiology and Oncology, Innsbruck Medical University, Anichstrasse 35, A-6020, Innsbruck, Austria.
authors: Thomas
authors: T
authors: Schmiedinger
authors: Georg F
authors: GF
authors: Vogel
authors: Oliver
authors: O
authors: Eiter
authors: Kristian
authors: K
authors: Pfaller
authors: Walter A
authors: WA
authors: Kaufmann
authors: Angelika
authors: A
authors: Flörl
authors: Karin
authors: K
authors: Gutleben
authors: Sabine
authors: S
authors: Schönherr
authors: Barbara
authors: B
authors: Witting
authors: Thomas W
authors: TW
authors: Lechleitner
authors: Hannes-L
authors: HL
authors: Ebner
authors: Thomas
authors: T
authors: Seppi
authors: Michael W
authors: MW
authors: Hess
mesh_terms: D018938:Caco-2 Cells; D020285:Cryoelectron Microscopy / Q000379:methods*; D005110:Extracellular Space / Q000737:chemistry; D005780:Gelatin / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D057139:Nanofibers / Q000737:chemistry
chemical_list: D005780:Gelatin
keywords: cell culture; cryofixation; cryosections; electrospinning; freeze- substitution; freeze-fracture replica labelling; gelatin nanofibres; high-pressure freezing; immunogold