Mice were anesthetized with isoflurane and transcardially perfused with PBS followed by 4% paraformaldehyde (PFA). Brains were removed from mice and subjected to post-fixation in 4% PFA, followed by dehydration in 30% sucrose. Twenty micrometer-thick sections from −3.14 mm −4.30 mm bregma were obtained using a cryostat (RRID: SCR_018061, Leica). To measure glial and neuronal cell loss in the mouse hippocampus sections were processed for immunostaining with antibodies against GFAP (1:1000), Iba1 (1:500), and neurofilament H non-phosphorylated (SMI-32) (RRID: AB_2564642, BioLegend, 1:500), MAP2 (RRID: AB_10693782, Cell Signaling Technology, 1:500). Subsequently, alexa488- (RRID: AB_2536161, Thermo Fisher Scientific Inc.,1:200) or Cy3-conjugated secondary antibodies (RRID: AB_2307443, Jackson ImmunoResearch Laboratories, Inc., 1:200) were used to visualize immunolabeling. Cell nuclei were visualized with DAPI (RRID: AB_2336790, Vector Laboratories, 1:5000). Fluorescence images in the CA1 or CA3 of the hippocampus were obtained from 2 sections per mouse using a laser scanning confocal microscope EZ-C1 (Eclipse TE2000U, Nikon). The fluorescence intensity of each region of the hippocampus/total area was analyzed using ImageJ software.
Ez c1 eclipse te2000u
Manufactured by Nikon
Sourced in Japan
The EZ-C1 (Eclipse TE2000U) is a confocal microscope system designed for advanced imaging applications. It features interchangeable laser modules, a motorized stage, and high-resolution imaging capabilities. The EZ-C1 is a versatile tool for researchers and scientists requiring detailed and precise microscopic analysis.
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Lab products found in correlation
2 protocols using ez c1 eclipse te2000u
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Immunohistochemical Analysis of Hippocampal Glia and Neurons
2
Quantifying Hippocampal Cell Loss
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Mice were anesthetized with iso urane and transcardially perfused with PBS followed by 4% paraformaldehyde (PFA). Brains were removed from mice and subjected to post-xation in 4% PFA, followed by dehydration in 30% sucrose. Twenty micrometer-thick sections from -3.14 mm -4.30 mm bregma were obtained using a cryostat (CM1950, Leica, Nussloch, Germany). To measure glial and neuronal cell loss in the mouse hippocampus sections were processed for immunostaining with antibodies against GFAP (1:1000), Iba1 (1:500), and neuro lament H non-phosphorylated (SMI-32) (801701, Biolegend, CA, USA 1:500), MAP2 (4542, Cell Signaling Technology, Tokyo, Japan 1:500). Subsequently, alexa488-(Thermo Fisher Scienti c Inc.,1:200) or Cy3-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc., PA, USA, 1:200) were used to visualize immunolabeling. Fluorescence images in the CA1 or CA3 of the hippocampus were obtained from 2 sections per mouse using a laser scanning confocal microscope EZ-C1 (Eclipse TE2000U, Nikon, Tokyo, Japan). The uorescence intensity of each region of the hippocampus/total area was analyzed using ImageJ software.
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