Immunocytochemical Analysis of Neural Cells

Immunocytochemical tests were carried out using an immunoassay kit (DAKO, EUA) according to the manufacturer’s protocol. For phenotypic characterization of primary cultures of neural cells, neurons were labeled with antibodies against the NeuN protein (anti-NeuN) (DAKO, EUA), astrocytes were labeled with antibodies against glial fibrillary acidic protein (anti-GFAP) (Thermo Scientific, EUA), and microglia were labeled with antibodies against adapter-ionized (AIP) calcium-binding molecule 1 (anti-IBA 1) (DAKO). The immunohistochemical pattern of cultures was obtained by calculating the percentage of labeled cells (NeuN, GFAP, and IBA 1) in relation to the total number of cells stained, as previously reported by Kádár et al. ^{( 6 (link) )}The cells were analyzed using an inverted microscope. Control wells not treated with the primary antibodies were included in the test.

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Souto A.D., Casseb S.M., de Melo K.F., Martins AJ F.i.l.h.o, & Franco E.C. (2023). Replication of Rocio virus in primary cultures of mouse neural cells. Einstein, 21, eAO0160.

Publication 2023

anti-GFAP anti-IBA 1

Antibodies Astrocytes Calcium Cells Gfap Immunoassay Microglia Microscope Neural Neurons Phenotypic Primary cells cultures Protein

Corresponding Organization : Universidade Federal do Pará

Other organizations : Instituto Evandro Chagas, Hospital Israelita Albert Einstein

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Percentage of labeled cells (NeuN, GFAP, and IBA 1) in relation to the total number of cells stained

control variables

Control wells not treated with the primary antibodies

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