Ix83 inverted microscope

Manufactured by Olympus

Sourced in Japan, United States, Germany, United Kingdom

The IX83 inverted microscope is a high-performance research-grade microscope designed for a variety of applications. It features a sturdy, stable frame and advanced optics to provide clear, high-resolution imaging. The IX83 is capable of various imaging modes, including brightfield, phase contrast, and fluorescence microscopy.

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Lab products found in correlation

Cellsens dimension software, by Olympus (13 mentions) Cellsens software, by Olympus (7 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (5 mentions) Aqua poly mount, by Polysciences (4 mentions) Orca flash 4.0 camera, by Hamamatsu Photonics (4 mentions) Cellsens, by Olympus (4 mentions) Ixon ultra 888 emccd camera, by Oxford Instruments (4 mentions) Permount, by Thermo Fisher Scientific (3 mentions) Senescence β galactosidase staining kit, by Cell Signaling Technology (3 mentions) Matlab, by MathWorks (3 mentions) Dragonfly 550, by Olympus (3 mentions) Transwell insert, by Corning (3 mentions) Visiscope confocal cell explorer system, by Zeiss (3 mentions) Fv1200 confocal system, by Olympus (2 mentions) μ slide 8 well glass bottom chamber, by Ibidi (2 mentions) Scmos fusion cameras, by Hamamatsu Photonics (2 mentions) Na 1.5 oil objective, by Hamamatsu Photonics (2 mentions) X cite led illumination system, by Excelitas (2 mentions) Uplanxapo, by Olympus (2 mentions) Te2000 u microscope, by Nikon (2 mentions)

Spelling variants of this product

Inverted microscope (2417 citations) IX83 microscope (347 citations) IX83 inverted fluorescence microscope (44 citations) IX83 inverted fluorescent microscope (13 citations) IX83 research inverted microscope (6 citations) IX83 inverted microscope system (6 citations) IX83) Inverted Spinning Disk Confocal Microscope (4 citations) FV1200MPE-IX83 inverted microscope (3 citations) IX83 P2ZF inverted microscope (2 citations) IX83 Inverted Light Microscope (2 citations)

274 protocols using ix83 inverted microscope

Visualizing C. elegans Touch Neuron Morphology

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mec‐7p::GFP was used to label touch neurons and mec‐7p::GFP::RAB‐3 was used to label synaptic vesicles in touch neurons. Well‐fed L4 animals were manually picked to synchronize the animals to regular NGM plates (40 worms/plate) and then transferred to NGM plates containing 50 μM FUDR (5‐fluoro‐2’‐deoxyuridine) on the first day of adulthood. Animals at the indicated days of adult life were mounted onto 2% agarose pad and immobilized with 30 μM muscimol (Sigma). Animals were then examined for neuronal morphology (blebbing and branching) or the distribution of GFP::RAB‐3 puncta using Zeiss LSM780 confocal microscope or Olympus IX83 inverted microscope. Each experiment was done in triplicate and results were compared to a control strain handled in parallel with the test strains. muIs71 (DAF‐16::GFP) was used to examine DAF‐16 intracellular localization. Well‐fed L4 animals were manually picked and mounted, immobilized and scored/imaged using Olympus IX83 inverted microscope.

Xu A., Zhang Z., Ko S., Fisher A.L., Liu Z, & Chen L. (2019). Microtubule regulators act in the nervous system to modulate fat metabolism and longevity through DAF‐16 in C. elegans. Aging Cell, 18(2), e12884.

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Confocal Imaging of Hydrated Biofilms

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The non-invasive confocal imaging of fully hydrated biofilms was carried out by means of a fixed-stage Ix83 Olympus inverted microscope coupled to an Olympus FV1200 confocal system (Olympus, Shinjuku, Tokyo, Japan). The objective lens was a ×63 water-immersion lens (Olympus). Specimens were stained at room temperature with LIVE/DEAD^® BacLight^TM Bacterial Viability Kit solution (L7012, Molecular Probes B. V., Leiden, The Netherlands). A staining time of 8 ± 1 min, in a 1:1 fluorochrome ratio was used to obtain the best fluorescence signal at the corresponding wavelengths (Syto9, 515–530 nm and propidium iodide, PI > 600 nm). At least three different and demonstrative locations of the discs were selected for the study. A z-series of scans (xyz) of 1 μm thickness (8 bits, 1024 × 1024 pixels) were analyzed thanks to the configuration of the CLSM control software. Image stacks were analyzed by using the Olympus^® software (Olympus^®). To quantify the biomass and cell viability within the biofilm, total fluorescent staining of the confocal micrographs was analyzed using an open source image analysis software (Fiji ImageJ) by measuring voxel intensities from two-channel images and, thus, calculating the percentage of the biomass and cell viability within the stacks [25 (link)].

Bueno J., Virto L., Toledano-Osorio M., Figuero E., Toledano M., Medina-Castillo A.L., Osorio R., Sanz M, & Herrera D. (2022). Antibacterial Effect of Functionalized Polymeric Nanoparticles on Titanium Surfaces Using an In Vitro Subgingival Biofilm Model. Polymers, 14(3), 358.

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In Vitro Cell Migration Assay

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Cells were grown in two-well silicone inserts (Ibidi 80209) in 24-well plates until confluent. The insert was then removed to form a 500 μm defined cell-free gap. The cells were then washed with PBS and given fresh media. Cell migration was imaged using an IX-83 Olympus inverted microscope at 0 h and 18 h after start of migration. Migration rate was calculated as the area covered over time.

Su S., Begum S, & Ezratty E.J. (2020). An IFT20 mechanotrafficking axis is required for integrin recycling, focal adhesion dynamics, and polarized cell migration. Molecular Biology of the Cell, 31(17), 1917-1930.

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Live/Dead Biofilm Imaging via CLSM

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Non-invasive confocal imaging of fully hydrated biofilms was carried out using a fixed-stage Ix83 Olympus inverted microscope coupled to an Olympus FV1200 confocal system (Olympus; Shinjuku, Tokyo, Japan). LIVE/DEAD^® BacLight™ Bacterial Viability Kit solution (Molecular Probes B. V., Leiden, The Netherlands) was used to stained the biofilms at room temperature. The fluorochromes were incubated (ratio 1:1) during 9 ± 1 min to obtain the optimum fluorescence signal at the corresponding wave lengths (Syto9: 515–530 nm; Propidium Iodide (PI): > 600 nm. The CLSM software was set to take a z-series of scans (xyz) of 1 μm thickness (8 bits, 1024 × 1024 pixels). Image stacks were analyzed by using the Olympus^® software (Olympus). Image analysis and live/dead cell ratio (i.e. the area occupied by living cells divided by the area occupied by dead cells) was performed with Fiji software (ImageJ Version 2.0.0-rc-65 / 1.52b, Open source image processing software).

Sánchez M.C., Ribeiro-Vidal H., Esteban-Fernández A., Bartolomé B., Figuero E., Moreno-Arribas M.V., Sanz M, & Herrera D. (2019). Antimicrobial activity of red wine and oenological extracts against periodontal pathogens in a validated oral biofilm model. BMC Complementary and Alternative Medicine, 19, 145.

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Quantifying Cell Adhesion Footprint

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The μ-Slide 8 Well Glass Bottom chamber (ibidi) was coated with recombinant human ICAM-1-Fc (5 μg/mL) at room temperature for 3 h. Differentiated MFN2 KD and control HL60 cells (2 × 10⁶ cells/mL) cells were incubated with CellTracker Orange CMRA (4 μM) at room temperature for 1 h. After 2 washes with PBS, cells were incubated with unconjugated mouse anti-human CD18 blocking Ab (TS1/18, 4 μg/mL) or mouse IgG1κ isotype control (4 μg/mL) at room temperature for 10 min. After 2 washes with PBS, cells were resuspended in PBS plus 1 μM Mn²⁺, added into the chamber, and centrifuged at 500 × g at room temperature for 5 min to induce spreading. Cells were fixed with 1% PFA at room temperature for 5 min and washed twice with PBS to remove unadhered cells. Total internal reflection fluorescence (TIRF) images (Fig. 4A) were acquired with an iX83 Olympus inverted microscope equipped with a SAFe Light module (Abbelight, includes four color lasers, λ = 405 nm, 488 nm, 532 nm, and 640 nm), sCMOS fusion cameras (Hamamatsu), and a 100× NA 1.5 oil objective. A TIRF incidence angle of θ = 70° was used. The area of cell footprint was quantified by the “analyzing particles” function in FIJI-ImageJ2^{58 (link)} (Fig. 4B).

Liu W., Hsu A.Y., Wang Y., Lin T., Sun H., Pachter J.S., Groisman A., Imperioli M., Yungher F.W., Hu L., Wang P., Deng Q, & Fan Z. (2021). Mitofusin-2 regulates leukocyte adhesion and β2 integrin activation. Journal of leukocyte biology, 111(4), 771-791.

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Actin Polymerization Assay in HL60 Cells

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Differentiated MFN2 KD and control HL60 cells (5 × 10⁵ cells/mL) were incubated with fMLP (100 nM) or PMA (100 nM) or vehicle control at room temperature for 20 min. Cells were fixed with 1% PFA at room temperature for 10 min, washed twice with intracellular staining perm wash buffer plus 5% goat serum, and stained with AF568-conjugated phalloidin (5 U/mL) in intracellular staining perm wash buffer plus 5% goat serum at room temperature for 30 min. After 2 washes with PBS, cells were added into μ-Slide 8 Well Glass Bottom chamber (ibidi) pre-coated with 0.01% Poly-l-lysine (at 4°C overnight) and centrifuged at 500 × g at room temperature for 5 min to let the cells settle and adhere. Epifluorescence images (Fig. 3A)were acquired by using an iX83 Olympus inverted microscope equipped with the SAFe Light module (Abbelight, includes four color lasers, λ = 405 nm, 488 nm, 532 nm, and 640 nm), sCMOS fusion cameras (Hamamatsu), and a 100× NA 1.5 oil objective. The phalloidin median fluorescence intensity (MFI), which reflects the actin polymerization, was quantified by the “analyzing particles” function in FIJI-ImageJ2^{58 (link)} (Fig. 3B).

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TUNEL Assay for Apoptosis-Induced DNA Damage

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To detect and quantify apoptosis-induced DNA damage, the DeadEnd™ fluorometric TUNEL assay kit (Promega Corporation, Madison, WI, USA) was used according to the manufacturer’s instructions. Here, 25,000 cells of melanoma cell line MEL-JUSO, SK-MEL-28, or MV3 were seeded on round 13-mm coverslips in a 12-well plate and treated with siCtrl or miR-101-3p mimic for 72 h. After washing the cells two times with PBS, they were fixed on the coverslips with 4% formaldehyde in PBS (pH 7.4) for 25 min at 4 °C. After removing the fixative solution, the cells were permeabilized with 0.2% Triton X-100 solution for 5 min and subsequently washed twice with PBS. To continue the treatments, coverslips were transferred to a light-shielded staining chamber. Cells were incubated in 100 µl of equilibration buffer for 10 min before incubation with 50 µl of rTdT buffer for at least 1 h at 37 °C. To stop the enzyme reaction, cells were incubated with 50 µl of 2 × SSC buffer for 15 min and thereupon washed three times with PBS. Finally, nuclear staining was performed using DAPI (1:10,000 in PBS) for 30 min before the coverslips were fixed on slides using Aqua-Poly/Mount (Polysciences). For staining analysis, an Olympus IX83 inverted microscope was used in combination with Olympus CellSens Dimension software (version 2.3, Olympus).

Lämmerhirt L., Kappelmann-Fenzl M., Fischer S., Meier P., Staebler S., Kuphal S, & Bosserhoff A.K. (2024). Loss of miR-101-3p in melanoma stabilizes genomic integrity, leading to cell death prevention. Cellular & Molecular Biology Letters, 29, 29.

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Periodic Acid-Schiff Staining Protocol

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Periodic acid-Schiff (PAS) staining was performed according to the manufacturer’s instructions (catalog no. ab150680; Abcam). Slides were mounted with Cytoseal 60 (catalog no. 83104; Thermo Scientific). Images were captured on an IX83 inverted microscope (Olympus) using a UC90 color charge-coupled-device (CCD) camera (Olympus).

Wells A.I., Grimes K.A, & Coyne C.B. (2022). Enterovirus Replication and Dissemination Are Differentially Controlled by Type I and III Interferons in the Gastrointestinal Tract. mBio, 13(3), e00443-22.

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Fluorescent Microparticle Imaging and Analysis

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Fluorescently labeled magnetic microparticles were imaged in optical-bottom 96-well plates on an IX83 inverted microscope (Olympus, Tokyo, Japan). Epifluorescence excitation was produced using an X-Cite LED illumination system (Excelitas, Wheeling, IL). An automated stage was employed to obtain 9 acquisitions per well using a pco.panda camera (PCO, Kelheim, Germany) and a 20x air objective (UPlanXApo, NA = 0.80, Olympus). Each acquisition consisted of a brightfield image to locate the microparticles and a fluorescent image (Cy3 filter cube, Edmond Optics, Barrington, NJ) to record the fluorescence intensity. Imaging analysis, including large fluorescent aggregate removal (e.g., dust/hair), calculation of the total microparticle pixel area and total microparticle fluorescence, was performed in Metamorph Advanced software (Molecular Devices, San Jose, CA), as previously described^{17 (link)}.

Macdonald P.J., Schaub J.M., Ruan Q., Williams C.L., Prostko J.C, & Tetin S.Y. (2022). Affinity of anti-spike antibodies to three major SARS-CoV-2 variants in recipients of three major vaccines. Communications Medicine, 2, 109.

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Immunofluorescence Staining of Melanoma Tissue

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Immunofluorescence stainings were performed on 5 µm patient-derived human cutaneous malignant melanoma tissue. Briefly, Antigen retrieval was carried out with Citrate-Buffer pH 6.7 for 20 min at 99 °C before sections were permeabilized with 1% Triton X-100 in PBS and blocked with 1% BSA/PBS. Primary antibody was added and incubated overnight at 4 °C. Afterwards sections were incubated with the respective secondary antibody 1:400 in PBS containing 0.5% Triton X-100. The antibodies used are listed in Supplementary Table 2. In the final step, nuclei were stained with DAPI (1:10,000 in 1% BSA/PBS, Sigma Aldrich), and sections were mounted on coverslips with Aqua Polymount (Polysciences). An Olympus IX83 inverted microscope was used for the analysis of the immunofluorescent staining.

Staebler S., Rottensteiner-Brandl U., El Ahmad Z., Kappelmann-Fenzl M., Arkudas A., Kengelbach-Weigand A., Bosserhoff A.K, & Schmidt S.K. (2024). Transcription factor activating enhancer-binding protein 2ε (AP2ε) modulates phenotypic plasticity and progression of malignant melanoma. Cell Death & Disease, 15(5), 351.

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