We present here an analysis of our own results obtained by fixing various tissues in zinc-ethanol-formaldehyde (ZEF). As compared with other fixation methods, fixation in ZEF was found to provide greater sensitivity for immunocytochemical reactions for many study antigens, in many cases also allowing thermal demasking of antigens to be avoided and giving higher image quality using fluorescence and confocal laser microscopy. However, studies of low molecular weight antigens showed diffusion of antigens from their initial locations. These data lead to the conclusion that fixation of specimens in ZEF has potential for use both in immunocytochemical studies, including use of fluorescence and confocal laser microscopy, and in general histological practice.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
O. S. Alekseeva, D. E. Korzhevskii, V. A. Otellin, et al., “Hypoxic preadaptation to nitrogen narcosis and heat shock proteins in cerebral cortex neurons in these states,” Dokl. Ros. Akad. Nauk., 425, No. 4, 553–555 (2009).
I. P. Grigor’ev, M. S. Vasilenko, E. G. Sukhorukova, and D. E. Korzhevskii, “Use of various antibodies to tyrosine hydroxylase to study the catecholaminergic systems of the brain in mammals,” Morfologiya, 138. No. 6, 60–63, (2010).
I. P. Grigor’ev, E. G. Gilerovich, E. S. Petrova, et al., “Catecholaminergic structures of the forebrain in rats after transient ischemia,” Region. Krovoobr. Microtsirk., 11, No. 2, 57–60 (2012).
I. P. Grigor’ev, M. A. Shklyaeva, O. V. Kirik, et al., “Distribution of alpha-tubulin in forebrain structures in rats,” Morfologiya, 143, No. 1, 7–10 (2013).
A. N. Zakurina, D. E. Korzhevskii, and N. G. Pavlova, “Placental failure – morphofunctional parallels,” Zh. Akush. Zhensk. Bolez., 59, No. 5, 51–55 (2010).
O. V. Kirik, T. D. Vlasov, and D. E. Korzhevskii, “The neural stem cell markers nestin and musashi-1 in telencephalon cells in rats after transient focal ischemia,” Morfologiya, 142, No. 4, 19–24 (2012).
O. V. Kirik, P. A. Voronchikhin, E. I. Chumasov, and D. E. Korzhevskii, “Use of reactions for synaptophysin to detect neuroepithelial bodies in the lungs in rats,” in: Babukhinskie Lectures in Orel [in Russian], Retinoidy Press, Moscow (2011), No. 32, pp. 70–71.
O. V. Kirik and D. E. Korzhevskii, “Vimentin in cells in the ependyma and subventricular proliferative zone in the rat telencephalon,” Kletoch. Tekhnol. Biol. Med., No. 4, 210–214 (2012).
O. V. Kirik, E. G. Sukhorukova, and D. E. Korzhevskii, “Calciumbinding protein Iba-1/AIF1 in brain cells in rats,” Morfologiya, 137, No. 2, 5–8 (2010).
E. A. Kolos, and D. E. Korzhevskii, “Studies of neurogenesis in spinal ganglia in rats using immunohistochemical differentiation markers,” in: Functional Interhemisphere Asymmetry and Plasticity in the Brain, Neurology Scientific Center, Moscow (2012), pp. 293–296.
D. E. Korzhevskii, I. P. Grigor’ev, and V. A. Otellin, “Use of dehydrating fixatives containing zinc salts in neurohistological studies,” Morfologiya, 129, No. 1, 85–86 (2006).
D. E. Korzhevskii, T. R. Drai, and S. V. Kostyukevich, “An immunocytochemical method for detection of EC (enterochromaffin) cells in the rat intestinal mucosal epithelium,” Morfologiya, 133, No. 1, 78–81 (2008).
D. E. Korzhevskii, O. V. Kirik, and G. V. Benzin, “A method for selective detection of proliferating cells in structures of the forming nervous system based on detection of phosphorylated histone H3,” Morfologiya, 142, No. 6, 69–72 (2012).
D. E. Korzhevskii, O. V. Kirik, and A. V. Gilyarov, “Rat brain cells containing esrin (cytovillin),” Morfologiya, 140, No. 6, 86–88 (2011).
D. E. Korzhevskii, O. V. Kirik, E. S. Petrova, and E. G. Sukhorukova, “Studies of cell proliferation in structures of the developing nervous system” in: Questions of Morphology in the 21st Century, DEAN Press, St. Petersburg (2012), No. 3, pp. 65–69.
D. E. Korzhevskii, O. V. Kirik, E. G. Sukhorukova, et al., “Studies of the spatial organization of astrocytes in the brain using confocal laser microscopy”, Morfologiya , 135, No. 3, 76–79 (2009).
D. E. Korzhevskii, O. V. Kirik, E. G. Sukhorukova, and T. D. Vlasov, “Structural organization of striatal microgliocytes after transient focal ischemia,” Morfologiya, 141, No. 2, 28–32 (2012).
D. E. Korzhevskii, M. V. Lentsman, O. V. Kirik, and V. A. Otellin, “Vimentin-immunopositive cells in the rat telencephalon after experimental ischemic stroke,” Morfologiya, 132, No. 5, 23–27 (2007).
D. E. Korzhevskii, V. A. Otellin, I. P. Grigor’ev, et al., “Immunocytochemical detection of neuronal NO synthase in rat brain cells,” Morfologiya, 132, No. 4, 77–80 (2007).
D. E. Korzhevskii, E. S. Petrova, O. V. Kirik, and V. A. Otellin, “Assessment of neuron differentiation during embryogenesis in rats using immunocytochemical detection of doublecortin,” Morfologiya, 133, No. 4, 7–10 (2008).
R. Lillie, Histopathologic Technic and Practical Histochemistry [Russian translation], Mir, Moscow (1969).
V. A. Nagornev, E. I. Chumasov, D. E. Korzhevskii, et al., “Current approaches to studies of the neural apparatus of arteries in atherosclerosis in diabetes mellitus,” Med. Akad. Zh., 10, No. 3, 19–27 (2010).
E. S. Petrova, “VImentin and glial fibrillary acidic protein in cells in ectopic neural transplants from neocortex,” Morfologiya, 139, No. 2, 22–26 (2011).
E. S. Petrova, “Astrogliogenesis in heterotopic allotransplants of embryonic neocortex in rats,” Byull. Eksperim. Biol., 151, No. 4, 191–195 (2011).
E. S. Petrova, T. D. Vlasov, E. A. Kolos, and D. E. Korzhevskii, “GABAergic neurons in the rat striatum in health and ischemic injury,” in: Functional Interhemisphere Asymmetry and Plasticity in the Brain, Neurology Scientific Center, Moscow (2012), pp. 347–349.
E. S. Petrova, E. N. Isaeva, and D. E. Korzhevskii, “Immunohistochemical studies of the differential of dissociated rat embryonic spinal cord and neocortex cells after allotransplantation into injured nerves,” ibid., pp. 344–347.
E. G. Sukhorukova, O. S. Alekseeva, O. V. Kirik, et al., “Comparative aspects of the structural organization of astrocytes in cortical layer I in the human and rat brain,” Zh. Evolyuts. Biokhim., 48, No. 3, 280–286 (2012).
E. G. Sukhorukova, O. V. Kirik, and D. E. Korzhevskii, “Use of an immunohistochemical method to detect microglia in the brain in paraffin sections,” Byull. Eksperim. Biol., 149, No. 6, 709–712 (2010).
E. I. Chumasov, P. A. Voronchikhin, and D. E. Korzhevskii, “Efferent innervation of vessels and bronchi in the lings in rats (an immunohistochemical study,” Morfologiya, 142, No. 4, 49–53 (2012).
E. I. Chumasov, E. S. Petrova, and D. E. Korzhevskii, “Immunomorphological analysis of the innervation of chromaffin cells in mammalian vascular and cardiac paraganglia,” Zh. Evolyuts. Biokhim., 47, No. 4, 325–331 (2011).
J. H. Beckstead, “A simple technique for preservation of fixation-sensitive antigens in paraffin-embedded tissues: addendum,” J. Histochem. Cytochem., 43, No. 3, 345 (1995).
L. Benerini-Gatta, M. Cadei, P. Balzarini, et al., “Application of alternative fixatives to formalin in diagnostic pathology,” Eur. J. Histochem., 56, 63–70 (2012).
Author information
Authors and Affiliations
Corresponding author
Additional information
Translated from Morfologiya, Vol. 143, No. 2, pp. 81–85, March–April, 2013.
Rights and permissions
About this article
Cite this article
Korzhevskii, D.E., Sukhorukova, E.G., Gilerovich, E.G. et al. Advantages and Disadvantages of Zinc-Ethanol-Formaldehyde as a Fixative for Immunocytochemical Studies and Confocal Laser Microscopy. Neurosci Behav Physi 44, 542–545 (2014). https://doi.org/10.1007/s11055-014-9948-8
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11055-014-9948-8