Fluoro-Jade B Staining of Degenerating Brain Cells

FJB was used to stain brain cells under degeneration [79 (link)]. Tissue 8 µm-thick sections were deparaffinized and hydrated. Slides were incubated in 80% EtOH-1% NaOH for 5 min, followed by 2 min in 70% ethanol and 2 min in distilled water. Sections were immersed in 0.06% K₂MnO₄ for 20 min, after distilled water washes, slides were incubated at 4 °C overnight in 0.001% Fluoro-Jade B (Millipore, AG310) previously diluted in 0.1% acetic acid [80 (link)]. Slides were dried, immersed in xylene and mounted with Eco-Mount (Biocare Medical, EM897L). Images were captured at 40× magnification on a Leica DM LS microscope with epifluorescence (Leica Microsystems, Wetzlar, Germany).

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Toral-Rios D., Patiño-López G., Gómez-Lira G., Gutiérrez R., Becerril-Pérez F., Rosales-Córdova A., León-Contreras J.C., Hernández-Pando R., León-Rivera I., Soto-Cruz I., Florán-Garduño B, & Campos-Peña V. (2020). Activation of STAT3 Regulates Reactive Astrogliosis and Neuronal Death Induced by AβO Neurotoxicity. International Journal of Molecular Sciences, 21(20), 7458.

Publication 2020

Fluoro-Jade B Eco-Mount Leica DM LS microscope

Acetic acid Brain Cells degeneration Ethanol Fluoro jade b Microscope Stain Tissue Xylene

Corresponding Organization : Instituto Nacional de Neurología y Neurocirugía

Other organizations : Instituto Politécnico Nacional, Center for Research and Advanced Studies of the National Polytechnic Institute, Hospital Infantil de México Federico Gómez, Research Institute of Molecular Pathology, Vienna Biocenter, Universidad Anáhuac, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Universidad Autónoma del Estado de Morelos, Universidad Nacional Autónoma de México

Top 5 similar protocols

Variable analysis

independent variables

Staining with FJB

dependent variables

Visualization of degenerating brain cells

control variables

Tissue thickness (8 µm)
Deparaffinization and hydration of tissue sections
Incubation in 80% EtOH-1% NaOH for 5 min
Incubation in 70% ethanol for 2 min
Incubation in distilled water for 2 min
Immersion in 0.06% K2MnO4 for 20 min
Incubation in 0.001% Fluoro-Jade B (FJB) diluted in 0.1% acetic acid at 4 °C overnight
Mounting with Eco-Mount
Imaging at 40× magnification on a Leica DM LS microscope with epifluorescence

controls

Positive control: Not specified
Negative control: Not specified

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