Ix83 inverted microscope

Manufactured by Hamamatsu Photonics

Sourced in United Kingdom

The IX83 inverted microscope is a modular and customizable research-grade microscope system designed for a wide range of applications. It features a sturdy and stable frame that supports various observation techniques, including brightfield, phase contrast, and fluorescence. The IX83 offers high-quality optics and a versatile design to accommodate a variety of sample types and experimental requirements.

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Lab products found in correlation

Orca er camera, by Hamamatsu Photonics (2 mentions) Orca flash 4.0 camera, by Hamamatsu Photonics (2 mentions) Mitomycin c, by Bio-Techne (1 mentions) Cellsens software, by Olympus (1 mentions) Ab179475, by Abcam (1 mentions) Alex fluor 594, by Jackson ImmunoResearch (1 mentions) F3648, by Merck Group (1 mentions) Bx63 upright microscope, by Olympus (1 mentions) Cellsens dimension, by Olympus (1 mentions) Orca flash 4.0 camera, by Olympus (1 mentions) Everbrite hardset mounting medium, by Biotium (1 mentions)

6 protocols using ix83 inverted microscope

Migrastasis: Cell Migration Assay

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Sterile 2-well culture inserts (Ibidi GmbH, Gräfelfing, Germany) were placed into 12 well culture plates. Cells were seeded at 50,000 cells into each side of the Ibidi insert then treated with designated siRNA for 24 h. Ibidi inserts were removed to create ‘cell-free gap’. Mitomycin C (1 µg/ml; Tocris, Abingdon, UK) was added to designated treatment conditions to prevent cell proliferation from confounding the cell migration results. Regions of interest (cell-free gap area) were immediately defined after Ibidi insert removal through CellSens software (Olympus, Essex, UK). Defined regions of interest were imaged at 0, 24, and 48 h. Images were collected on an Olympus IX83 inverted microscope using a 4x objective and captured using an Orca ER camera (Hamamatsu, Hertfordshire, UK). Images were then processed, and cell-free area was quantified using Fiji ImageJ (http://imagej.net/Fiji/Downloads; Method S4). % Gap closure was calculated as follow: % gap closure = ((gap area at 0 h−gap area at × h) ÷ gap area at 0 h) × 100

Sritangos P., Pena Alarcon E., James A.D., Sultan A., Richardson D.A, & Bruce J.I. (2020). Plasma Membrane Ca2+ ATPase Isoform 4 (PMCA4) Has an Important Role in Numerous Hallmarks of Pancreatic Cancer. Cancers, 12(1), 218.

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Microscopic Imaging Techniques

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Still phase contrast images were taken on an EVOS FL microscope using the 10x and 20x objective. Videos were captured on an Olympus IX83 inverted microscope using a 20x objective and captured using an Orca ER camera (Hamamatsu, UK) through MMI Cell tools software (MMI, Switzerland).

Kisby T., de Lázaro I., Stylianou M., Cossu G, & Kostarelos K. (2021). Transient reprogramming of postnatal cardiomyocytes to a dedifferentiated state. PLoS ONE, 16(5), e0251054.

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Time-Lapse TIRF Imaging of Root Epidermal Cells

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Roots of 7-d-old seedlings were imaged with an Olympus IX83 inverted microscope equipped with a Cell^TIRF module and Hamamatsu EM-CCD C9100-13 camera, using OLYMPUS Uapo N 100×/NA 1.49 Oil TIRF objective at 1.6× magnification. Single channel imaging was done sequentially with the mentioned time interval. Time-lapse imaging in roots was done in the epidermal cells of the transition zone in TIRF mode (Johnson et al., 2020 (link)).

Narasimhan M., Gallei M., Tan S., Johnson A., Verstraeten I., Li L., Rodriguez L., Han H., Himschoot E., Wang R., Vanneste S., Sánchez-Simarro J., Aniento F., Adamowski M, & Friml J. (2021). Systematic analysis of specific and nonspecific auxin effects on endocytosis and trafficking. Plant Physiology, 186(2), 1122-1142.

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Visualizing Integrin Dynamics in HT-1080 Cells

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Non-transfected or GFP-VANGL2 transfected HT-1080 cells were plated on vitronectin or fibronectin treated coverslip dishes. Cells were left untreated or treated with MnCl₂ and fixed using 4% paraformaldehyde/PBS at room temperature or ice-cold methanol at −20°C. Washed cells were blocked using PBS, 5% normal donkey serum, and 0.1% Triton X-100 for 1 h at room temperature. Cells were incubated with LAMP1 (1:100, D2D11 XP; Cell Signaling Technology), fibronectin (1:100, F3648, Millipore Sigma), paxillin (1:100, AHO0492, Thermo Fisher), or integrin αv (1:100, ab179475, Abcam) primary antibody in block for 1 h at room temperature, washed twice with PBS, and incubated an additional 1 h in the dark with Donkey anti-Rabbit or anti-mouse Alex Fluor 594 (1:500, 711–585-152/715–585-150; Jackson ImmunoResearch Laboratories). Cells were washed three times with PBS. NucBlue Fixed Cell DAPI Stain (Thermo Fisher Scientific) was used to label nuclei. Imaging was performed using a 40x oil objective (NA = 1.3) or 20x dry objective (NA = 0.75) and an Olympus IX83 inverted microscope equipped with a Hamamatsu Orca Flash 4.0 camera. For some images, maximum intensity projections were generated from z-stack slices (0.5 μm).

Jessen T.N, & Jessen J.R. (2018). VANGL2 protein stability is regulated by integrin αv and the extracellular matrix. Experimental cell research, 374(1), 128-139.

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Immunofluorescence Imaging of Cellular Proteins

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Cells were grown on glass coverslips in 6-well culture dishes, fixed in 4% paraformaldehyde for 15 min at room temperature, washed in icecold PBS, and permeabilized for 12 min in permeabilization buffer (0.2 % Triton X-100, 1% BSA in PBS). Unspecific fluorescence was blocked by a 30 min incubation in PBS plus 2% BSA, followed by incubation with primary antibody for 1

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h at room temperature, three washes in 2% BSA blocking buffer, and incubation with secondary antibodies diluted in 2% BSA blocking buffer) for 45 min, washing in PBS containing 2% BSA, a 5 min incubation with (4',6-Diamidino-2-Phenylindole, Dihydrochloride) (DAPI) for nuclear staining, extensive washing in PBS and mounting in N-propyl-gallate mounting media (2% w/v in PBS/glycerin). Fluorescence images were captured on a fully motorized Olympus BX63 upright microscope with an Olympus DP72 color, 12.8-megapixel, 4.140 × 3.096-resolution camera and with a fully motorized and automated Olympus IX83 Inverted microscope with a Hamamatsu ORCA-Flash 4.0 camera (C11440-22CU). The software used was Olympus CellSens dimension, which is able to do deconvolution on captured z stacks, and images were processed for publication using Adobe Photoshop CS6.

Malinda R.R., Zeeberg K., Sharku P.C., Ludwig M.Q., Pedersen L.B., Christensen S.T, & Pedersen S.F. (2020). TGFβ Signaling Increases Net Acid Extrusion, Proliferation and Invasion in Panc-1 Pancreatic Cancer Cells: SMAD4 Dependence and Link to Merlin/NF2 Signaling. Frontiers in Oncology, 10, 687.

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Visualizing Nematode Worms via Microscopy

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Adult worms were placed on agarose pads with a 10 μL drop of S-basal and covered with a coverslip. The worms were imaged on an Olympus IX83 inverted microscope equipped with a Hamamatsu Orca-Flash 4.0 camera, using a 10× UPLSAPO objective and managed by Olympus CellSens software. For DAPI staining, worms were fixed in ice-cold methanol for 5 min before being mounted in EverBrite™ Hardset Mounting medium (Biotium 23004).

Jensen H.H., Frantzen M.T., Wesseltoft J.L., Busuioc A.O., Møller K.V., Brohus M., Duun P.R., Nyegaard M., Overgaard M.T, & Olsen A. (2023). Human calmodulin mutations cause arrhythmia and affect neuronal function in C. elegans. Human Molecular Genetics, 32(12), 2068-2083.

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