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Ix71 system

Manufactured by Olympus
Sourced in Japan

The Olympus IX71 system is a research-grade inverted microscope designed for advanced imaging and analysis applications. It features a modular design that allows for customization to meet specific experimental requirements. The IX71 system provides a stable and reliable platform for a variety of microscopy techniques, including fluorescence, brightfield, and phase contrast imaging.

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6 protocols using ix71 system

1

Ovarian Sample Extraction during Estrous Cycle

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To retrieve ovarian samples from adult mice (8 weeks old) during each of the four stages of the estrous cycle, we performed the vaginal smear tests to identify the specific estrous cycle stages. Briefly, we placed the tip of a plastic pipette filled with 40 μl of phosphate buffered saline after (PBS; Hyclone, USA) into the mouse vagina, and gently flushed the vagina 3–5 times. Vaginal contents (cells and vaginal fluids) from each mouse were collected into a PCR tube and then smeared onto a glass slide. The evaluation of the vaginal smear images was performed under a microscope IX71 system (Olympus, Japan). Proestrus was characterized by the mostly nucleated and some cornified epithelial cells in the smear; estrus by cornified squamous epithelial cells without visible nuclei; metestrus by the predominance of leucocytes and a few nucleated epithelial and/or cornified squamous epithelial cells; and diestrus was characterized by a predominance of leukocytes [24 (link), 25 ]. The reader is directed to further citations for a full description [24 (link), 25 ]. Vaginal smear tests were performed at 08:00 and 20:00 each day. When the stages were identified, the mice were sacrificed via cervical dislocation and their ovaries were collected.
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2

Mitochondrial Protein Localization in Myoblasts

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Patient-derived myoblasts were seeded onto 4-well culture slides and were maintained at 37 °C under humidified atmosphere of 5 % CO2. After 3 days in culture, cells were fixed, permeabilized, and blocked according to standard immunocytochemical protocol. Primary antibody probing was performed at room temperature for 2 h. Secondary antibody probing was performed with 2.5 μg/mL Alexa Fluor 568 (Molecular Probes) at room temperature for 1 h. Mitochondria were co-stained with 0.25 μg/mL MitoTracker Green (Molecular Probes). Stained cells were observed under a fluorescent microscope (IX71 System; Olympus). Primary antibodies used were as follows: 2.5 μg/mL anti-MT-CO1 (Molecular Probes), 2.5 μg/mL anti-COX4 (Molecular Probes).
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3

Cytochemical COX Staining of SH-SY5Y Cells

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Cytochemical COX staining was performed as follows: Briefly, undifferentiated and differentiated SH-SY5Y WT and SH-SY5Y ρ0 were stained with COX reaction buffer (pH 5.5; 100 mM sodium acetate, 0.1% MnCl2, 0.001% H2O2, 10 mM diaminobenzidine) at 37 °C for 1 h, followed by subsequent incubation with 1% CuSO4 at 37 °C for 5 min. Stained samples were observed under a microscope (IX71 System; Olympus).
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4

Quantitative Immunohistochemical Analysis of Mucosal Mast Cells

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Immunohistochemical staining was performed according to the procedure described in previous reports [21 (link), 31 (link)–33 ]. Briefly, one hour after the post-OIT non-heated OVA challenge, the proximal colon was excised, fixed by immersion in 4% paraformaldehyde and then embedded in OCT compound. Cutted sections (30 μm) were exposed to antiserum against mMCP-1, a marker of mouse mucosal mast cells (1:5,000; Moredun Scientific, Scotland, UK) and then incubated with Cy3-conjugated sheep anti-donkey IgG (1:200; Jackson Immunoresearch Laboratories). The immunostained sections were examined using a fluorescence microscope (IX71 system; Olympus, Tokyo, Japan) with a U-MWIG3 filter set (Olympus) and photographed using an Olympus digital camera (DP70; Olympus). The histological staining was quantitatively analyzed using ImageJ software.
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5

Immunofluorescence Staining of Mouse Colon

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The colonic sections were treated with a rat IgG anti-mouse CD11c antibody, rat IgG anti-mouse CD4 antibody, rabbit IgG anti-rat CGRP antibody, or goat IgG anti-mouse CCL21 antibody for 12–18 h at 4 °C. After the incubation, the sections were incubated with a suitable secondary antibody for 2 h at RT. The sections were observed using a confocal laser scanning microscope (LSM700; Carl Zeiss Japan, Tokyo, Japan) or a fluorescence microscope (IX71 system; Olympus, Tokyo, Japan) with a U-MWIG3 filter set (Olympus) and photographed using an Olympus digital camera (DP70; Olympus).
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6

Cell Migration and Invasion Assays

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In vitro cell migration was performed using Thincert Cell Culture Insert For 24 Well Plates (Greiner Bio‐One). Briefly, a volume of 600 µl of cell suspension (1 × 104 cells) in DMEM serum‐free medium was added into the upper chambers. A volume of 600 µL DMEM medium with 10% FBS without antibiotic was added in the lower chamber to induce cell migration. After 24 h incubation, the medium was removed, and migrated cells were fixed with 4% paraformaldehyde and stained using 0,5% crystal violet solution. The non‐invading cells were removed from the insert's upper surface using a cotton swab, and five random fields were photographed using the inverted microscope IX71 system (Olympus). Images were later processed, quantified and analysed using ImageJ software.
In vitro cell invasion was conducted using BioCoat Matrigel Invasion Chamber assay (Corning). The invasion chamber was removed from the freezer and rehydrated with DMEM medium at 37°C. DMEM was added to the insert's interior and the bottom of wells 2 h before plating the cells. The following steps were performed as described above for the migration assay. All the experiments were performed in triplicate.
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