Axiovert 200 m microscopy

Manufactured by Zeiss

The Axiovert 200 M is an inverted microscope system designed for advanced microscopy applications. It features a high-quality optical system, a motorized focus, and a modular design that allows for the integration of various accessories and imaging techniques. The Axiovert 200 M is a versatile laboratory tool suitable for a wide range of microscopy-based research and analysis.

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Lab products found in correlation

Apotome, by Zeiss (2 mentions) Goat anti mouse alex 595, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Axiovert 200 (1017 citations) Axiovert (339 citations) Axiovert 200 M microscopy system (3 citations)

4 protocols using axiovert 200 m microscopy

Evaluating FUS and RNAP CTD Droplet Localization

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To evaluate GFP-RNAP CTD localization, samples were prepared by adding 0.3 μM GFP-CTD to phase separated 300 μM FUS SYGQ in 50 mM MES/Bis-Tris 150 mM NaCl pH 5.5, 50 μM FUS ΔSYGQ in 20 mM sodium phosphate 150 mM NaCl pH 7.4, or 5 μM MBP-FUS FL with 0.015 mg/mL TEV protease in 20 mM sodium phosphate 150 mM NaCl pH 7.4. Samples containing MBP-FUS FL were incubated with TEV protease for 20 min prior to addition of GFP-RNAP CTD.
Samples were spotted onto a glass coverslip and droplet formation was evaluated by imaging with differential interference contrast or using the FITC channel for fluorescence on an Axiovert 200M microscopy (Zeiss). For experiments with 5 μM MBP-FUS FL and variants, the samples were incubated with 0.01 mg/mL in-house TEV protease for 20 min before visualization.

Murthy A.C., Tang W.S., Jovic N., Janke A.M., Seo D.H., Perdikari T.M., Mittal J, & Fawzi N.L. (2021). Molecular interactions contributing to FUS SYGQ LC/RGG phase separation and co-partitioning with RNA polymerase II heptads. Nature structural & molecular biology, 28(11), 923-935.

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Cellular Localization of GCS and GM130

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For cellular localization studies, mouse skin fibroblasts were seeded onto chamber slides one day prior to fixation with 4% paraformaldehyde for 1–2 hrs at room temperature (RT). The fixed cells were washed 3 times with 1X PBS for 10 min each. Then cells were fixed with ice cold methanol (−80°C) for 5 min and washed 3 times with 1X PBS for 10 min each. Cells were then permeabilized with 0.3% Triton X-100/PBS for 1–2 hr RT. Non-specific antibody binding was blocked with 1.5% BSA/1.5% Milk/0.1% Gelatin in 1X PBS 1–2 hrs at RT. The cells were incubated with primary antibodies, rabbit anti-mouse GCS (17 kD) and Alexa Fluor 555 Mouse anti-GM130 (Golgi marker) in blocking solution overnight at 4°C, followed by anti-rabbit conjugated Alexa-Fluor 488 for 4 hrs at RT. The cells were washed with PBS+0.01% Tween 20. Cell nuclei were stained with DAPI. Fluorescence signals were visualized by Zeiss Axiovert 200 M microscopy equipped with an Apotome. Pearson Correlation Coefficients were determined for individual cells using Zeiss Axiovision co-localization software.

Barnes S., Xu Y.H., Zhang W., Liou B., Setchell K.D., Bao L., Grabowski G.A, & Sun Y. (2014). Ubiquitous Transgene Expression of the Glucosylceramide-Synthesizing Enzyme Accelerates Glucosylceramide Accumulation and Storage Cells in a Gaucher Disease Mouse Model. PLoS ONE, 9(12), e116023.

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Staurosporine-Induced Apoptosis Imaging

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To initiate apoptosis, cells were treated with 2 μM staurosporine and incubated another 60–90 min before imaging. Imaging was conducted on an Axiovert 200 M microscopy (Zeiss) equipped with a 75 W xenon-arc lamp and 20× objective lens (NA = 0.75, air) and a 14-bit CoolSnap HQ2 cooled CCD camera (Photometrics), driven by open source Micro-Manager software. For a typical experiment, images were recorded at 1 minute intervals.

Li Y., Forbrich A., Wu J., Shao P., Campbell R.E, & Zemp R. (2016). Engineering Dark Chromoprotein Reporters for Photoacoustic Microscopy and FRET Imaging. Scientific Reports, 6, 22129.

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Immunohistochemistry of alpha-synuclein

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Frozen sagittal brain sections from WT and 9V/null mice were fixed (4% PFA, 30 min) and permeabilized (0.3% Triton X-100, 20 min). The sections were treated with proteinase K (10 μg/mL) for 7 min in TNB buffer (TSA kit, PerkinElmer NEL705A001) to remove soluble αSyn from tissue sections. The sections were incubated with anti-mouse αSyn monoclonal antibody (Abcam 3H9, 1:200 dilutions) overnight at 4°C. Fluorescent signals were detected by goat anti-mouse Alex 595 (Invitrogen). Fluorescence signals were visualized and captured by Zeiss Axiovert 200 M microscopy equipped with an Apotome.

Dai M., Liou B., Swope B., Wang X., Zhang W., Inskeep V., Grabowski G.A., Sun Y, & Pan D. (2016). Progression of Behavioral and CNS Deficits in a Viable Murine Model of Chronic Neuronopathic Gaucher Disease. PLoS ONE, 11(9), e0162367.

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