title: VIS2FIX: a high-speed fixation method for immuno-electron microscopy.
abstract: Immuno-transmission electron microscopy (TEM) is the technique of choice for high-resolution localization of proteins in fixed specimen. Here we introduce 2 novel methods for the fixation of sections from cryo-immobilized samples that result in excellent ultrastructural preservation. These high-speed fixation techniques, both called VIS2FIX, allow for a reduction in sample preparation time from at least 1 week to only 8 h. The methods were validated in immuno-TEM experiments on THP-1 monocytes, human umbilical vein endothelial cells (HUVECs) and Madin-Darby canine kidney (MDCK-II) cells. The fixation and retention of neutral lipids is demonstrated, offering unique prospects for the application of immuno-TEM in the lipidomics field. Furthermore, the VIS2FIX methods were successfully employed in correlative fluorescence and electron microscopy.
authors: Molecular Biophysics, Department of Physics and Astronomy, Utrecht University, Princetonplein 1, NL-3508 TA Utrecht, The Netherlands. M.A.Karreman@uu.nl
authors: Matthia A
authors: MA
authors: Karreman
authors: Elly G
authors: EG
authors: van Donselaar
authors: Hans C
authors: HC
authors: Gerritsen
authors: C Theo
authors: CT
authors: Verrips
authors: Arie J
authors: AJ
authors: Verkleij
mesh_terms: D000818:Animals; D002460:Cell Line; D004285:Dogs; D006801:Humans; D007150:Immunohistochemistry; D008055:Lipids / Q000737:chemistry; D016253:Microscopy, Immunoelectron / Q000379:methods*; D013997:Time Factors; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D008055:Lipids