VIS2FIX: A High-Speed Fixation Method for Immuno-Electron Microscopy
Traffic, Vol. 12, No. 8, p.806-. ISSN 1398-9219.
Karreman, M.A.; van Donselaar, E.G.; Gerritsen, H.C.; Verrips, C.T.; Verkleij, A.J.; Biomolecular Imaging; Soft Condensed Matter and Biophysics; Sub Soft Condensed Matter; Sub Molecular Biophysics; Sub Cell Biology; Sub Biomolecular Imaging
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.