Axio observer inverted microscope

Manufactured by Zeiss

Sourced in Germany, Japan, United States

The Axio Observer is an inverted microscope designed for a wide range of applications in life science research. It features a sturdy, ergonomic design and provides high-quality optics for brightfield, phase contrast, and fluorescence imaging. The Axio Observer is equipped with a motorized stage and supports a variety of sample holders, enabling efficient and precise observation and analysis of specimens.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (13 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (6 mentions) Axiocam mrm camera, by Zeiss (6 mentions) Axiovision software, by Zeiss (6 mentions) Apotome system, by Zeiss (5 mentions) Compact light source hxp 120c, by Zeiss (5 mentions) Hoechst 33342, by Thermo Fisher Scientific (4 mentions) Alexa fluor 555, by Thermo Fisher Scientific (4 mentions) Zen software, by Zeiss (4 mentions) Lsm 780, by Zeiss (4 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions) Lsm 700 confocal module, by Zeiss (3 mentions) Alexa fluor 488, by Abcam (3 mentions) Dig labeling reagents and rna polymerases, by Roche (3 mentions) Triton x 100, by Merck Group (3 mentions) Thermomixer c shaker, by Eppendorf (3 mentions) Pa1 41331, by Agilent Technologies (3 mentions) Prism 6, by GraphPad (3 mentions) Prolong with dapi, by Thermo Fisher Scientific (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions)

182 protocols using axio observer inverted microscope

Visualizing Shigella Infection in Amoebae

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Amoebae seeded on 4-well μ-slides (Ibidi) were infected as above with GFP (green fluorescent protein)-expressing S. sonnei 53G and GFP-expressing S. flexneri M90T. Amoebae were then washed with low-fluorescence (LF) medium (41 (link)) and placed on ice, and extracellular bacteria were detected with sera against S. sonnei (Remel agglutinating sera) or S. flexneri 5a (Public Health England [PHE]) in 2% BSA in LF medium for 30 min. Amoebae were washed with ice-cold LF medium, followed by anti-rabbit Cy3 (Jackson ImmunoResearch) in 2% BSA in PAS for 30 min. Amoebae were washed with LF medium before being overlaid with agarose and immediately imaged on a Zeiss Axio Observer inverted microscope.
For neutral red staining, amoebae were incubated in 125 μM neutral red in LF medium for 20 min at room temperature (RT). Amoebae were washed in LF medium before addition of bacteria harboring GFP and centrifuged briefly (2 min, 1,000 × g). Amoebae were overlaid with agarose and immediately imaged on a Zeiss Axio Observer inverted microscope.

Watson J., Jenkins C, & Clements A. (2018). Shigella sonnei Does Not Use Amoebae as Protective Hosts. Applied and Environmental Microbiology, 84(9), e02679-17.

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Immunofluorescence Assay for GLI1

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SKGT4 with and without siRNA transfection were seeded into 4-well Nunc Lab-Tek II Chamber Slides (Thermo Fisher Scientific, Waltham, MA). After reaching 90% confluence, the cells were fixed in acetone at −20 °C for 7 minutes, washed three times with 1xPBS, and then incubated with primary antibodies for GLI1. Subsequently, the cells were incubated with AlexaFluor goat anti-rabbit secondary antibodies. The cells were counterstained with Hoechst 33342 (Thermo-Fisher Scientific), embedded with paraffin, and observed with a Zeiss Axio Observer Inverted Microscope (Zeiss, Oberkochen, Germany) for fluorescent imaging.

Ng C.K., Ma K., Cheng Y., Miyashita T., Harmon J.W, & Meltzer S.J. (2019). Krüppel-like Factor 5 Promotes Sonic Hedgehog Signaling and Neoplasia in Barrett's Esophagus and Esophageal Adenocarcinoma. Translational Oncology, 12(11), 1432-1441.

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Syncytia Formation Assay for HA Mutants

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BHK-21 cells were transfected with 1 µg of pCAGGS plasmid encoding the NL09 wild-type or escape mutant HA proteins using Lipofectamine (Invitrogen, USA) and Plus reagent (Invitrogen, USA). At 24 h posttransfection, HA-expressing cells were washed once with PBS and treated with 2.5 µg/mL of TPCK-treated trypsin (Sigma-Aldrich, USA) for 10 min. Afterward, the cells were exposed to pH adjusted PBS for 5 min at 37 °C. The cells were neutralized by complete growth media (MEM, supplemented with GlutaMAX™ Supplement, 5% FBS and PS) and incubated for 2 h at 37 °C to allow syncytia formation. The cells were fixed and stained with the Hema3Stat Pak (Fisher Scientific, USA) according to the manufacturer’s protocol. Syncytia were visualized and photographed using a Zeiss Axio Observer inverted microscope with an attached digital camera (Zeiss, Germany).

Lee C.Y., Raghunathan V., Caceres C.J., Geiger G., Seibert B., Cargnin Faccin F., Gay L.C., Ferreri L.M., Kaul D., Wrammert J., Tan G.S., Perez D.R, & Lowen A.C. (2023). Epistasis reduces fitness costs of influenza A virus escape from stem-binding antibodies. Proceedings of the National Academy of Sciences of the United States of America, 120(17), e2208718120.

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Spinning-Disk Confocal Microscopy Workflow

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All images were acquired with a Mariana system (Intelligent Imaging Innovations, Denver, CO) based on a Zeiss Axio-Observer inverted microscope (Carl Zeiss Microimaging, Inc., Thornwood, NY) equipped with a CSU-X1 spinning-disk confocal unit (Yokogawa Electric, Tokyo, Japan), a piezo-driven Z-translation, and linear encoded X&Y translations and controlled with SlideBook V5.0 (Intelligent Imaging Inc., Denver, CO). Excitation wavelengths were 405, 488, 561, and 640 nm (lasers were from Cobolt, Solna, Sweden); the emission filters were 452/45, 525/50, 607/36, and 680-nm long-pass (Semrock, Rochester, NY). Exposure times and laser settings for each experiment are outlined above.

Salgado E.N., Upadhyayula S, & Harrison S.C. (2017). Single-Particle Detection of Transcription following Rotavirus Entry. Journal of Virology, 91(18), e00651-17.

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Immunostaining of Stem Cell Markers

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For microscopy analyses, the hBMSCs were cultured on glass coverslips and treated according to experimental groups described earlier. After 3 days, cells were washed with PBS, fixed in PBS-paraformaldehyde 4% for 1 h and permeabilized in PBS containing 0.2% Triton-X 100 and 1% BSA at 37°C for 1 h and stained with primary antibodies diluted according to the manufacturer’s instructions: mouse monoclonal OCT4 (ab-59545, Abcam; 1:100 dilution) and mouse monoclonal NANOG (1E6C4—sc-293121, Santa Cruz Biotechnology; 1:100 dilution) for 1 h at room temperature. After washing with PBS, the cells were stained with secondary antibody rabbit anti-mouse IgG H+L Alexa Fluor 488 (Invitrogen) for 1 h at room temperature. Cells were washed with PBS and coverslips were mounted on glass slides using Fluoroshield with DAPI (Sigma) and then viewed on Zeiss Axio Observer inverted microscope with Apotome.2 (Zeiss, Germany). The quantification of NANOG and OCT4 was performed as previously described [25 (link)].

Assis R.I., Wiench M., Silvério K.G., da Silva R.A., Feltran G.D., Sallum E.A., Casati M.Z., Nociti FH J.r, & Andia D.C. (2018). RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation. PLoS ONE, 13(12), e0207873.

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Glucocorticoid Receptor Immunofluorescence

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The ± ENZ-treated cells (VCaP: 100 000 per well; 22Rv1: 40 000 per well) were cultured in charcoal stripped medium on 8-well chamber slides (Ibidi GmbH, #80826). Before staining, the cells were treated with EtOH or 100 nM Dex for 1 h at 37°C. For fluorescent staining, the cells were fixed with 4% paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.4 (PB) for 20 min, and washed with PB. The fixed cells were permeabilized for 15 minutes with 0.1% Triton X-100 and 1% BSA, blocked with 1% BSA for 20 min, and incubated o/n at 4°C with anti-GR antibody (1:500). After washing with PB, the cells were incubated for 2 h with Alexa Fluor 647 labeled secondary antibody (1:500), and nuclei were labeled with 4′,6-diamidino-2-phenylindole (DAPI, 1 μg/ml) (Sigma-Aldrich, #D8417). The fluorescent images were obtained with a Zeiss Axio Observer inverted microscope (40× NA 1.3 oil objective) equipped with a Zeiss LSM 800 confocal module (Carl Zeiss Microimaging GmbH). Image processing was performed using ZEN 2.5 (blue) software (Carl Zeiss Microimaging GmbH), and the images were quantified with ImageJ software (National Institutes of Health).

Helminen L., Huttunen J., Tulonen M., Aaltonen N., Niskanen E.A., Palvimo J.J, & Paakinaho V. (2023). Chromatin accessibility and pioneer factor FOXA1 restrict glucocorticoid receptor action in prostate cancer. Nucleic Acids Research, 52(2), 625-642.

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Immunocytochemical Staining of Mouse Astrocytes

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Immunocytochemical studies were performed on mouse cultured astrocytes adherent to coverslips (Schildge et al., 2013 ) and stained with Abcam primary antibodies (GAPDH ab8245; GFAP ab7260) and fluorescent-tagged secondary antibody (mouse, AlexaFluor 555 or rabbit, AlexaFluor 488; Thermofisher Scientific, France). Cells were imaged using a 20 × Plan-Apochromat (aperture 0.8)/63x EC Plan-Neofluar oil-immersion lens (aperture 1.3) of a Zeiss Axio Observer inverted microscope (Carl Zeiss, Jena, Germany) equipped with an AxioCam MRm camera.

Bénit P., Pelhaître A., Saunier E., Bortoli S., Coulibaly A., Rak M., Schiff M., Kroemer G., Zeviani M, & Rustin P. (2017). Paradoxical Inhibition of Glycolysis by Pioglitazone Opposes the Mitochondriopathy Caused by AIF Deficiency. EBioMedicine, 17, 75-87.

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Immunofluorescence Imaging of Tight Junctions

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Cells, 1.0 × 10⁵, were seeded and grown as a monolayer on 8‐well chambered slides (LAB‐TEK) for 3 weeks. Cells were washed with PBS and fixed with one of the following steps: (1) preextracted with 0.2% TritonX/PBS on ice followed by 1% paraformaldehyde/PBS for 15 min at RT; (2) fixed directly with 1% paraformaldehyde/PBS for 15 min at RT; (3) fixed directly with 100% methanol for 5 min on ice; and (4) fixed directly with 100% Acetone for 5 min on ice. Cells were permeabilized with 0.5% Tween/PBS and blocked in 0.1% Tween/casein/PBS before primary antibody staining. Tissue postfixation was done with 4% paraformaldehyde/PBS and mounted with Prolong Gold antifade reagent with DAPI (Life Technologies). Images were obtained using a spinning disk BD CARVII Confocal Imager (BD Biosystems, San Jose, CA) on a Zeiss Axio Observer inverted microscope (Carl Zeiss Microscopy, Thornwood, NY) at 63X objective lens controlled by MetaMorph imaging software (Molecular Devices, Sunnyvale, CA). Image Z resolution was further optimized with Volocity software (PerkinElmer, Waltham, MA). For non‐Z‐stacked higher resolution images, a Leica SP5 inverted laser confocal micorscope was used. Antibodies used: Anti‐claudin 3(#ab15102, AbCam, Cambridge, MA), anti‐claudin4 (#329400, Life Technologies), Anti‐human CD137 (#552533, BDBioscience), phalloidin‐Alexa 647(A22287, Life Technologies).

Gusti V., Bennett K.M, & Lo D.D. (2014). CD137 signaling enhances tight junction resistance in intestinal epithelial cells. Physiological Reports, 2(8), e12090.

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Biofilm Inhibition by Nanoparticles

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Two Gram-positive bacteria species, S. aureus and S. epidermidis, were used for the biofilm fluorescence assay. Biofilms were allowed to form on glass coverslips submerged with 1 mL of TSB in a 24-well plate at 37°C for 24 hours. Then, the supernatant containing non-adhered cells was removed and the coverslips were washed with sterile PBS. Existing biofilms were incubated for another 24 hours with TSB supplemented with different concentrations of Ag/AuNRs, making the final concentrations: 0, 15 and 25 µg/mL. Biofilms were fixed using a 4% paraform-aldehyde solution for 30 minutes at room temperature and then were washed three times with PBS. After that, 5-(4, 6-dichlorotriazinyl) aminofluorescein was added into the wells and the samples were allowed to incubate with shaking for 2 hours at room temperature. The coverslips were washed three times in PBS and inverted onto a cover glass. Images were acquired using a Zeiss Axio Observer Inverted microscope (Carl Zeiss Meditec AG, Jena, Germany).

Zhang J., Wang M, & Webster T.J. (2018). Silver-coated gold nanorods as a promising antimicrobial agent in the treatment of cancer-related infections. International Journal of Nanomedicine, 13, 6575-6583.

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Visualizing phagosomal maturation and hyphal growth

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Live cell imaging was performed to monitor phagosomal maturation by by using a Zeiss Axio Observer inverted microscope (Carl Zeiss Inc., Thornwood, NY) equipped with a confocal laser system housed in the Duke University Light Microscope Core Facility (LMCF) or by using an UltraVIEW Vox live cell imaging system equipped with a CSU-1 spinning disk scanner, housed in the Institute of Medical Sciences at the University of Aberdeen. Images obtained were analyzed and processed with the Zeiss LSM 510 version 4.2 (Carl Zeiss Inc., Thornwood, NY) or with the Volocity software (PerkinElmer), repectively. Phagosome maturation was monitored by measuring cathepsin B activity by employing the Magic Red Cathepsin B assay kit (ImmunoChemistry) following the manufacturer’s instructions. Hoechst 33342 was used to stain macrophage nuclei.
Time lapse analyses for hyphal growth of wild-type and cnaBΔ mutants were conducted by using a Zeiss Axio Observer Z1 microscope system (Carl Zeiss Inc., Thornwood, NY) with an Opto-electronically motorized XY stage, Pecon XL S1 incubator, and Coolsnap ES2 high-resolution CCD camera, housed in the Duke University Light Microscopy Core Facility (LMCF).
Transmission electron microscopy for spores, wild-type hyphae, wild-type yeast, and cnbRΔ yeast-locked mutants, was performed as described previously (Li et al., 2011 (link)).

Lee S.C., Li A., Calo S., Inoue M., Tonthat N.K., Bain J.M., Louw J., Shinohara M.L., Erwig L.P., Schumacher M.A., Ko D.C, & Heitman J. (2015). Calcineurin orchestrates dimorphic transitions, antifungal drug responses, and host-pathogen interactions of the pathogenic mucoralean fungus Mucor circinelloides. Molecular microbiology, 97(5), 844-865.

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