S Gull1, I Ingrisch1, S Tausch1, O W Witte1, S Schmidt2. 1. Hans Berger Department of Neurology, Jena University Hospital, 07747 Jena, Germany. 2. Hans Berger Department of Neurology, Jena University Hospital, 07747 Jena, Germany. Electronic address: Silvio.Schmidt@med.uni-jena.de.
Abstract
BACKGROUND: Golgi-Cox staining is a powerful histochemical approach which has been used extensively to visualize the morphology of neurons and glia. However, its usage as a first-choice method is hindered by its uncertain nature, diminished consistency and lengthy staining duration. The FD Rapid GolgiStain™ Kit (FD Neurotechnologies, Inc., USA) has been developed by employing the Golgi-Cox approach. It is a simple, reliable and reproducible way of performing Golgi impregnation for the analysis of neuronal morphology. NEW METHOD: We report here simple modifications to the manufacturer's protocol which enable reproducible and reliable staining of glial cells. RESULTS: Exposure of brain tissue to 4% paraformaldehyde (PFA) during perfusion followed by postfixation with 8% glutaraldehyde in 4% PFA led to only glial cells being stained, whereas in the absence of postfixation both neurons and glia were stained with unclear morphology. Additionally, we found that impregnation at 26°C±1 was critical to attain uniform staining. COMPARISON WITH EXISTING METHOD: Our modified Golgi-Cox approach is consistent and reproducible and affords uniform glial staining throughout the brain. CONCLUSION: As this protocol stains only a small percentage of cells, it is suitable for the analysis of individual cells.
BACKGROUND: Golgi-Cox staining is a powerful histochemical approach which has been used extensively to visualize the morphology of neurons and glia. However, its usage as a first-choice method is hindered by its uncertain nature, diminished consistency and lengthy staining duration. The FD Rapid GolgiStain™ Kit (FD Neurotechnologies, Inc., USA) has been developed by employing the Golgi-Cox approach. It is a simple, reliable and reproducible way of performing Golgi impregnation for the analysis of neuronal morphology. NEW METHOD: We report here simple modifications to the manufacturer's protocol which enable reproducible and reliable staining of glial cells. RESULTS: Exposure of brain tissue to 4% paraformaldehyde (PFA) during perfusion followed by postfixation with 8% glutaraldehyde in 4% PFA led to only glial cells being stained, whereas in the absence of postfixation both neurons and glia were stained with unclear morphology. Additionally, we found that impregnation at 26°C±1 was critical to attain uniform staining. COMPARISON WITH EXISTING METHOD: Our modified Golgi-Cox approach is consistent and reproducible and affords uniform glial staining throughout the brain. CONCLUSION: As this protocol stains only a small percentage of cells, it is suitable for the analysis of individual cells.
Authors: Matheus de Castro Fonseca; Bruno Henrique Silva Araujo; Carlos Sato Baraldi Dias; Nathaly Lopes Archilha; Dionísio Pedro Amorim Neto; Esper Cavalheiro; Harry Westfahl; Antônio José Roque da Silva; Kleber Gomes Franchini Journal: Sci Rep Date: 2018-08-13 Impact factor: 4.379
Authors: Ákos Menyhárt; Rita Frank; Attila E Farkas; Zoltán Süle; Viktória É Varga; Ádám Nyúl-Tóth; Anne Meiller; Orsolya Ivánkovits-Kiss; Coline L Lemale; Írisz Szabó; Réka Tóth; Dániel Zölei-Szénási; Johannes Woitzik; Stephane Marinesco; István A Krizbai; Ferenc Bari; Jens P Dreier; Eszter Farkas Journal: J Cereb Blood Flow Metab Date: 2021-08-24 Impact factor: 6.960