Paraffin-embedded mouse or human glioma surgical specimens were baked for 3 h at 60 °C and subjected to microwave treatment in citrate buffer for antigen retrieval. Next, the sections were incubated with 3% H2O2 for 15 min, blocked with 5% BSA for 1 h and incubated with primary antibodies (Ki67, YANK2, p-p70S6K T389) overnight at 4 °C and secondary antibodies for 30 min. H&E staining was used for histological examination of the brains of tumor-bearing mice. The whole regions of each tumor specimen were observed with an Olympus imaging system microscope and collected by NDP software. view2. The IOD scores of YANK2 in human GBM or mouse GBM models were quantified by software image J under ×400 magnified microscope with an Olympus imaging system microscope in the whole regions of each tumor specimen.
Microscope imaging system
Manufactured by Olympus
Sourced in Japan
The Microscope Imaging System from Olympus is a high-performance optical microscope designed for advanced imaging applications. It features a modular and versatile design that can be customized to meet the specific needs of various research and industrial applications. The core function of this system is to provide users with the ability to capture detailed, high-quality images of microscopic samples, enabling in-depth analysis and observation.
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Lab products found in correlation
Matrigel, by BD (4 mentions)
Cell culture insert, by BD (3 mentions)
Matrigel matrix, by BD (2 mentions)
Af647 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg af488, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Transwell cell culture insert, by BD (1 mentions)
Ab178846, by Merck Group (1 mentions)
Ab182422, by Abcam (1 mentions)
Sc 271430, by Abcam (1 mentions)
Ba0412, by Boster Bio (1 mentions)
Mouse or rabbit igg1, by Santa Cruz Biotechnology (1 mentions)
Costar chamber, by Corning (1 mentions)
30 protocols using microscope imaging system
1
Immunohistochemical Profiling of Glioma
2
Wound Healing Assay with A549 Cells
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After plating A549 cells onto 35 mm dishes, they were cultured until reaching confluence. At 48 h after the transfection with siRNAs, a wound was created in the cell layer using a plastic pipette tip (P100), and then at 8 h after wound healing, the length of the wound was subsequently measured using a microscope imaging system (Olympus, Tokyo, Japan).
3
Immunofluorescence Assay of Cell Markers
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For the immunofluorescence assay, cells were fixed, permeabilized at 4 °C for 30 min, incubated with the indicated antibodies (rabbit anti-ACE2 pAb, Proteintech, 21115-1-AP, 1:50; mouse anti-ASGPR1 mAb, Sino Biological, 10773-MM02, 1:50; rabbit anti-ASGPR1 mAb, Sino Biological, 10773-R011, 1:50; and mouse anti-KREMEN1 mAb, homemade, 1:100) at 4 °C overnight. The cells were washed twice with PBS, stained with labeled secondary antibodies (goat anti rabbit IgG/AF488, Thermo Fisher, A11034; goat anti mouse IgG/AF647, Thermo Fisher, A21236; goat anti human IgG/AF647, Thermo Fisher, A21455; goat anti mouse IgG/AF594, Thermo Fisher, A11005; each at 1:200) at 4 °C for 2 h, and subjected to 4′6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich) staining for observation under a microscope imaging system (Olympus, Japan).
4
Wound Healing Assay with Detroit562 Cells
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After Detroit562 cells were plated onto 35 mm dishes, they were cultured to confluence. At 24 h after we wounded the cell layer using a plastic pipette tip (P200), we measured the distance of the wound using a microscope imaging system (Olympus, Tokyo, Japan).
5
Transwell Invasion Assay for Cell Migration
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For invasion assays, Transwell cell culture inserts were used (pore size, 8-µm; Becton Dickinson and Company). Detroit 562 cells (1×104) were seeded at 37°C into the upper Matrigel pre-coated chamber (1:50, Becton Dickinson and Company) with serum-free medium at 4°C for 30 min, and the lower chamber was filled with human fibroblast conditioned medium containing 10 nM EGF as an adhesive substrate. The cells were incubated for 24 h at 37°C. The upper chamber was the fixed with 100% methanol for 10 min and stained with Giemsa for 20 min. The number of invading cells was assessed using a microscope imaging system (Olympus Corporation).
6
Wound Healing Assay on Cultured Cells
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After the Sawano cells were plated onto the 35 mm dishes, they were cultured to confluence. At 24 h we wounded the cell layer using a plastic pipette tip (P200), and measured the length of the wound by using a microscope imaging system (Olympus, Tokyo, Japan).
7
Wound Healing Assay with Sawano Cells
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After the Sawano cells were plated onto the 35 mm dishes, they were cultured to confluence. At 24 h we wounded the cell layer using a plastic pipette tip (P200), and measured the length of the wound by using a microscope imaging system (Olympus, Tokyo, Japan).
8
Wound Healing Assay with A253 Cells
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After the A253 cells were plated onto 35 mm dishes, they were cultured to confluence. At 24 h, we wounded the cell layer using a plastic pipette tip (P200), and then measured the length of the wound by using a microscope imaging system (Olympus, Tokyo, Japan).
9
Oxidative Stress Evaluation Protocol
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A ROS assay kit (Genmed Scientifics Inc.) was used to detect ROS levels. The kit contained a fluorogenic probe (DCFH-DA), which could react with ROS. Fluorescence was detected using a spectrofluorometer (excitation, 520 nm; emission, 490 nm). All procedures were performed according to the manufacturer’s instructions. The fluorescence intensity was observed using a microscope imaging system (Olympus America, United States).
Malondialdehyde (MDA) is a reliable indicator of lipid peroxidation while superoxide dismutase (SOD) is an endogenous scavenger of the reactive oxygen species (ROS). All procedures were performed in accordance with the manufacturer’s instructions provided along with the kits (Beyotime Biotechnology Institute, Nantong, China). The results are illustrated as the MDA concentration (nmol/mg protein) and SOD activity (U/mg).
Malondialdehyde (MDA) is a reliable indicator of lipid peroxidation while superoxide dismutase (SOD) is an endogenous scavenger of the reactive oxygen species (ROS). All procedures were performed in accordance with the manufacturer’s instructions provided along with the kits (Beyotime Biotechnology Institute, Nantong, China). The results are illustrated as the MDA concentration (nmol/mg protein) and SOD activity (U/mg).
10
Whole Mount In Situ Hybridization
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Whole mount in situ hybridization was performed as described previously [18 (link)]. Digoxigenin (DIG)-labeled antisense riboprobes (trypsin, lfabp, and ifabp) were synthesized by in vitro transcription reactions. DIG-labeled riboprobes were detected using alkaline phosphatase-conjugated anti-DIG antibodies; the hybridization signals were developed using NBT/INT solution as a substrate (Roche). Embryos were imaged by using Olympus microscope imaging system.
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