Selected article for: "bovine serum and confocal microscope"
Author: William Binning; Aja E. Hogan-Cann; Diana Yae Sakae; Matthew Maksoud; Valeriy Ostapchenko; Mohammed Al-Onaizi; Sara Matovic; Wei-Yang Lu; Marco A. M. Prado; Wataru Inoue; Vania F. Prado
Title: Chronic hM3Dq signaling in microglia ameliorates neuroinflammation in male mice
  • Document date: 2020_1_28
    • ID: ely200x3_26
    Snippet: Immunocytochemistry was done on microglia plated on poly-D-lysine-coated glass coverslips at a density of ~1×104 cells per well. Cells were obtained from three independent cultures of three different litters to confirm HA expression. Cells were fixed with 4% paraformaldehyde (PFA) in PBS. After permeabilization with 0.5% Triton X-100 and blocking with 5% bovine serum albumin (BSA)/0.1% Triton X-100, cultures were incubated with primary antibodie.....
    Document: Immunocytochemistry was done on microglia plated on poly-D-lysine-coated glass coverslips at a density of ~1×104 cells per well. Cells were obtained from three independent cultures of three different litters to confirm HA expression. Cells were fixed with 4% paraformaldehyde (PFA) in PBS. After permeabilization with 0.5% Triton X-100 and blocking with 5% bovine serum albumin (BSA)/0.1% Triton X-100, cultures were incubated with primary antibodies (anti-HA, anti-Iba1, and anti-GFAP) overnight at 4 °C. Cells were then washed with PBS and incubated for 1 hour with secondary antibodies (Alexa Fluor 555 goat anti-mouse and Alexa Fluor 633 goat anti-rabbit, 1:500) and Hoechst (1:2000) . Coverslips were then mounted on glass slides using Immumount (Thermo Fisher) medium and imaged using an FV1000 confocal microscope (Olympus) equipped with a 20x/0.75, 30x/1.05 or 60x/1.35 objective. Images were analyzed using ImageJ (NIH, Bethesda, MD) Cell Counter plugin. author/funder. All rights reserved. No reuse allowed without permission.

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