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Axiovert 200

Manufactured by Leica
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Sourced in Israel

The Axiovert 200 is an inverted microscope designed for laboratory applications. It provides high-quality imaging and observation capabilities for a variety of sample types.

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Lab products found in correlation

Abc kit, by Vector Laboratories (1 mentions) Dab kit, by Vector Laboratories (1 mentions) Anti nestin, by Merck Group (1 mentions) Anti collagen 1, by Merck Group (1 mentions) Anti ki67, by Merck Group (1 mentions) Anti fibronectin, by Merck Group (1 mentions) Anti collagen 4, by Merck Group (1 mentions) Anti sma, by Merck Group (1 mentions) Fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions) Anti cd90, by Abcam (1 mentions) Anti cd105, by BioLegend (1 mentions) Anti sm22α, by GeneTex (1 mentions) Anti pdgfrα, by Thermo Fisher Scientific (1 mentions) Anti c kit, by R&D Systems (1 mentions) Dmi6000b, by Zeiss (1 mentions) Femtojet injector, by Eppendorf (1 mentions) Alexa fluor 488 antibody, by Thermo Fisher Scientific (1 mentions) Anti acetylated tubulin antibody, by Merck Group (1 mentions) Discovery v8, by Zeiss (1 mentions) Lsm 510 duo, by Zeiss (1 mentions)

6 protocols using axiovert 200

1

Transwell Assay for Cell Migration

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Transwell assays were conducted using the HTS FluoroBlok Multiwell Insert System (Corning) with an 8.0 µm pore size. The bottom surface of the membrane was coated with laminin (10 µg/ml) for 3 hr at 37°C. Membranes were then blocked for 1 hr using 2% polyvinylpyrolidone. The bottom well was filled with 500 µl media and 1 × 104 cells were added to the top well. Cells were allowed to transmigrate for 16 hr before the bottom side of the membrane was fixed with 4% PFA. In order to quantify the number of cells that had transmigrated, the fixed sample was permeabilised using 0.1% TX-100 for 5 min at room temperature and nuclei were stained using Hoechst at a concentration of 0.1 µg/ml. Cells were imaged using epifluorescence imaging on an inverted microscope (Zeiss Axiovert 200 or Leica DMIL) fitted with a 10x/0.22 NA objective and standard filters; 4 regions of each filter were imaged. The ‘Analyze Particles’ function of ImageJ was used to count cells within each field. Data from each independent experiment were normalized to the average number cells that were counted on the underside of each filter for triplicate controls.
Patkunarajah A., Stear J.H., Moroni M., Schroeter L., Blaszkiewicz J., Tearle J.L., Cox C.D., Fürst C., Sánchez-Carranza O., Ocaña Fernández M.D., Fleischer R., Eravci M., Weise C., Martinac B., Biro M., Lewin G.R, & Poole K. (2020). TMEM87a/Elkin1, a component of a novel mechanoelectrical transduction pathway, modulates melanoma adhesion and migration. eLife, 9, e53308.
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2

Immunophenotyping of Aortic Cell Populations

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Aortic sections were immunostained with anti-laminin γ1 (Abcam), anti-nestin (Sigma), anti-SMA (Sigma), anti-SM22α (GeneTex), anti-fibronectin (Millipore), anti-collagen I (Sigma), anti-collagen IV (Millipore), anti-CD90 (Abcam), anti-CD105 (Biolegend), anti-PDGFRα (eBiosciences), anti-c-Kit (R&D), and anti-Ki67 (Millipore) antibodies overnight at 4°C. For fluorescent staining, sections were incubated with appropriate fluorescent secondary antibodies (Invitrogen) for 1 hour at room temperature. Due to strong autofluorescence in the green channel of aortic tissue, Alexa-647 (artificially colored in green) rather than Alexa-488 was used. For DAB staining, sections were incubated with biotinylated secondary antibodies for 1 hour at room temperature, followed by ABC kit (Vector) and DAB Kit (Vector), according to the manufacturer’s instructions. After mounting, the sections were examined and photographed with Zeiss Axiovert 200 or Leica confocal microscope.
Yao Y., Norris E.H, & Strickland S. (2014). The cellular origin of laminin determines its role in blood pressure regulation. Cellular and molecular life sciences : CMLS, 72(5), 999-1008.
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3

Targeted Gene Silencing in Embryos

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A dsRNA microinjection of a single 8-cell embryo blastomere, leading to specific clonal interference with gene function, was performed as described previously [34 (link), 35 , 47 (link)] using the FemtoJet injector (Eppendorf), under an inverted Leica DMI6000B or Zeiss Axiovert 200 microscope equipped with Leica or Eppendorf micromanipulators, respectively. Microinjections were carried out in micro-drops of M2 medium, after which the embryos were cultured as described above.
Filimonow K., Saiz N., Suwińska A., Wyszomirski T., Grabarek J.B., Ferretti E., Piliszek A., Plusa B, & Maleszewski M. (2019). No evidence of involvement of E-cadherin in cell fate specification or the segregation of Epi and PrE in mouse blastocysts. PLoS ONE, 14(2), e0212109.
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4

Immunostaining and Pronephros Analysis

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For immunostaining, embryos were fixed for 1-2h in MEMFA. The staining was performed as described before 136 . Cilia were detected with a monoclonal anti-acetylated tubulin antibody (Sigma, T7451) in combination with an anti-mouse secondary Alexa Fluor 488 antibody (Invitrogen, A-11001). Pictures were taken with a ZEISS LSM 510 DUO with inverted microscope Axiovert 200 and a 63x LCl-Plan Neofluar objective and a Leica SP8 inverse FALCON and a 63x HC PL APO CS2 objective. Ciliation was determined in Centrin-RFP-positive cells and the area of their acetylated tubulin staining was measured on maximum intensity projections with ImageJ 2.0.0.
For pronephros analysis a fluorescein-labeled Lycopersicon esculentum lectin (1:100 dilution; Vector Laboratories) was used for visualization. A SteREO Discovery.V8 from Zeiss and Zen2011 Blue Edition was used for imaging. Pronephros size was measured with the line tool of ImageJ 2.0.0 and the ratio of injected versus uninjected side calculated 6 .
Getwan M., Hoppmann A., Schlosser P., Grand K., Song W., Diehl R., Schroda S., Heeg F., Deutsch K., Hildebrandt F., Lausch E., Köttgen M., & Walz G. (2020). CRISPR/Cas9 targeting Ttc30a mimics ciliary chondrodysplasia with polycystic kidney disease.
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5

iPSC Pluripotency Marker Profiling

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ESC marker expression in iPSCs was assessed by immunofluorescence staining with primary antibodies against Oct3/4 (Santa Cruz Biotechnology, Santa Cruz, CA) and SSEA-1 (Cellular Signaling Technology, Beverly, MA). Corresponding fluorescein-labeled secondary antibodies raised in donkeys were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich). Images were acquired using a Leica Axiovert 200 inverted fluorescence microscope (Leica, Bannockburn, IL).
Xiang M., Lu M., Quan J., Xu M., Meng D., Cui A., Li N., Liu Y., Lu P., Kang X., Wang X., Sun N., Zhao M., Liang Q., Le L., Wang X., Zhang J, & Chen S. (2019). Direct in vivo application of induced pluripotent stem cells is feasible and can be safe. Theranostics, 9(1), 290-310.
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6

Immunofluorescence Analysis of Xenograft Tumors

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Immunofluorescence staining of xenograft tumors was performed on 6-μm paraffin tissue sections. After deparaffinization and rehydration, sections were treated with antigen retrieval buffer (DAKO) in a steamer for 20 min. Slides were incubated with 10 % normal goat serum (Cell Signaling) and with primary antibody/rabbit anti-GR H-300 (Santa Cruz, 1:100) and mouse anti-Ki67 (Dako, 1:50) diluted in PBS overnight at 4 °C. Slides were then incubated with secondary antibody/anti-mouse conjugated to Alexa 488 and anti-rabbit conjugated to Alexa Fluor 647 (Cell Signaling, 1:1,000). Sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, diluted 1:1,000 in PBS) (Invitrogen) to visualize nuclei and mounted with Fluoromount mounting medium (Sigma). Immunofluorescence staining was imaged using the AMG EVOS-FL microscope or Leica-Axiovert 200. Arbitrary false coloring of images was implemented using ImageJ software. Images were captured at same intensity and exposure for all conditions within each cell line. Any contrast/brightness changes were made consistently across images. For GR IF enumeration (VCaP xenografts), five representative fields were captured at ×40 magnification, and the numbers of GR-expressing and nuclear (overlapping with DAPI) cells were counted.
Isikbay M., Otto K., Kregel S., Kach J., Cai Y., Vander Griend D.J., Conzen S.D, & Szmulewitz R.Z. (2014). Glucocorticoid Receptor Activity Contributes to Resistance to Androgen-Targeted Therapy in Prostate Cancer. Hormones & Cancer, 5(2), 72-89.
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