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Axiovert s100 microscope

Manufactured by Zeiss
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Sourced in Germany

The Axiovert S100 is a high-quality inverted microscope designed for a variety of laboratory applications. It features a sturdy and stable construction, allowing for precise optical performance. The microscope is equipped with a range of objectives and illumination options to accommodate different samples and magnification requirements.

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Lab products found in correlation

Axiocam mrm camera, by Zeiss (3 mentions) Pluronic f27, by Thermo Fisher Scientific (2 mentions) Rhod2 am dye, by Thermo Fisher Scientific (2 mentions) Fluo 4 am, by Thermo Fisher Scientific (2 mentions) Axiocam icc3 camera, by Zeiss (2 mentions) Triton x 100, by Merck Group (2 mentions) Axiovision 4, by Zeiss (2 mentions) Dyecycle green, by Thermo Fisher Scientific (1 mentions) Retiga exi, by Zeiss (1 mentions) Seakem me agarose, by Lonza (1 mentions) Plan apo vc 20 objective, by Nikon (1 mentions) Oxotremorine m, by Santa Cruz Biotechnology (1 mentions) Cell clock cell cycle assay, by Biocolor (1 mentions) Trypan blue dye, by Corning (1 mentions) Dulbecco modified eagle medium (dmem), by Lonza (1 mentions) Mem non essential amino acids, by Lonza (1 mentions) Plan apochromat 63x 1.4 oil, by Zeiss (1 mentions) Lyve 1 ab, by Abcam (1 mentions) Hepes buffer, by Thermo Fisher Scientific (1 mentions) Image pro plus software, by Media Cybernetics (1 mentions)

48 protocols using axiovert s100 microscope

1

4T1-GFP Cells Interaction with MIRB Nanoparticles

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On day -2, 4T1-GFP cells were seeded in triplicate in a 6-well plate (3 × 104 cells/well). On day -1, MIRB nanoparticles were added to the culture medium (25 μg/ml, overnight incubation). On day 0, culture medium was removed, cells were washed 3 times with PBS and replenished with full medium. Fluorescence microscopy was performed to confirm the intracellular localization of MIRB particles, and 7.5 × 104 control J774 macrophages were added to MIRB-labeled 4T1-GFP cells. Cells were next imaged for 2 days using fluorescence microscopy (AxioVert S100 microscope, Zeiss). Exposure times were kept constant during experiments. After fluorescence imaging, cells were rinsed 3 times with PBS, fixed with paraformaldehyde 4% (Sigma-Aldrich, Bornem, Belgium), and washed 3 times with PBS containing 0.1% Triton X-100 (Sigma-Aldrich, Bornem, Belgium) for membrane permeabilization. Fixed cells were next incubated with a PBS solution containing 1% potassium ferrocyanide (Sigma-Aldrich, Bornem, Belgium)/1% HCl for 15 minutes. After Perls' Prussian blue staining of iron, cells were washed with PBS and brightfield pictures were taken (AxioVert S100 microscope, Zeiss).
Danhier P., Deumer G., Joudiou N., Bouzin C., Levêque P., Haufroid V., Jordan B.F., Feron O., Sonveaux P, & Gallez B. (2017). Contribution of macrophages in the contrast loss in iron oxide-based MRI cancer cell tracking studies. Oncotarget, 8(24), 38876-38885.
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2

Quantifying Collagen Degradation in HAECs

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siRNA transfected HAECs, labeled with a live cell nuclear dye (HOESCHT 33342), were trypsinized, resuspended in complete growth medium, and seeded on collagen I-coated coverslips supplemented with DQ-Collagen-I (25 µg/ml). Once the HAECs had adhered to the coverslips (20 min), cells were treated with select experimental compounds [VEGF-A165 (25 ng/ml), GM6001 (10 µmol/L; MedChem Express) or appropriate vehicles] for 4 hours. Following treatment, HAECs were briefly fixed in 4% paraformaldehyde (15 min) then imaged using a Zeiss Axiovert S100 microscope; immediate imaging post-fixation optimized visualization of fluorescent products derived from of DQ-collagen degradation. For these studies, collagen degradation was quantified as the average fluorescence intensity per cell for a minimum of 100 cells per experiment using ImageJ software. In some experiments, following imaging for collagen degradation, HAECs were washed, permeabilized in 0.1% triton, blocked in 0.3% BSA and stained with an anti-cortactin antibody (#05–180, clone 4F11; Millipore). Co-localization of the FITC DQ-Col-I degradation products and of anti-cortactin-staining in cortactin-rich circular structures (rosettes) was visualized with a Zeiss Axiovert S100 microscope.
MacKeil J.L., Brzezinska P., Burke-Kleinman J., Craig A.W., Nicol C.J, & Maurice D.H. (2019). A PKA/cdc42 Signaling Axis Restricts Angiogenic Sprouting by Regulating Podosome Rosette Biogenesis and Matrix Remodeling. Scientific Reports, 9, 2385.
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3

Estrous Cycle Determination via Vaginal Cytology

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Vaginal slides were dried for at least five minutes, then placed in a fixing solution (Rapid Fixx, Shandol) for one minute. Slides were placed in Harris Haematoxylin for 1 minute, then rinsed in tap water for 1 minute. Excess stain was removed with minimal amounts of water and dehydrated in 70% ethanol for 1 minute. Slides were submerged in 5% Eosin solution for 1 minute. Slides were rinsed again in tap water to remove excess stain and dehydrated in ascending alcohol. Finally, slides were cleared using xylenes and mounted using toluene-based media. Following staining slides were examined under a Zeiss AxioVert S100 microscope at 10x magnification to determine the phase of estrous (Fig 2A–2D).
Larocque J.C., Gardy S., Sammut M., McBey D.P, & Melling C.W. (2022). Sexual dimorphism in response to repetitive bouts of acute aerobic exercise in rodents with type 1 diabetes mellitus. PLoS ONE, 17(9), e0273701.
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4

Pollen Staining and Imaging Protocol

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Fresh pollen was collected from each plant, immediately fixed in ethanol: acetic acid (3:1 ratio) solution, and later rehydrated through an ethanol series for size measurements and staining. To visualize nuclei, fixed pollen grains were stained with DyeCycle Green (Thermo Fisher Scientific, Cat no. V35004) at a final concentration of 1 μM in water, incubated in dark for 30 min and washed with sterile water. Stained pollen was then imaged on a Zeiss Axiovert S100 microscope equipped with HBO 50 Illuminator and Chroma #41017 filter set (470/40 excitation, 495 long-pass dichroic, 525/50 emission) using Qimaging (Retiga ExI) camera.
Zhou L., Vejlupkova Z., Warman C, & Fowler J.E. (2021). A Maize Male Gametophyte-Specific Gene Encodes ZmLARP6c1, a Potential RNA-Binding Protein Required for Competitive Pollen Tube Growth. Frontiers in Plant Science, 12, 635244.
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5

Quantitative Analysis of Chemotactic Cell Migration

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1% BSA coated 6-well plates (In Vitro Scientific, California) were used. The assay was performed as previously described (13 (link)). Briefly, 0.5% SeaKem ME agarose (Lonza) in 50% each of PBS and RPMI without phenol red was allowed to solidify in wells. Three 1 mm diameter wells were carved out at 2 mm distance between each other. Vehicle (DMSO) or Ent treated (100 μM for 2 h at 37°C) cells were stained with CellTracker Red CMPTX dye according to manufacturer’s protocol (Life Technologies, Thermo Fisher Scientific). Cells were washed and re-suspended in RPMI without phenol red and 10 μl (5 × 104 cells) of vehicle or Ent-treated cells were added to the extreme wells and were allowed to migrate towards the center well containing 10 μl of either fMLP (100 nM) or LTB4 (250 nM). Fluorescence microscopy of samples was performed using a 1x objective lens (Zeiss Axiovert S100 microscope) and images of the same field of view were acquired every 45 seconds for a total period of 60 min. Image sequences from under-agarose movies were used to track 20 or more cells per condition in each experiment. Each image sequence was loaded on Image J and cells were followed using the Manual Tracking plugin. The output files containing the × and y coordinates were then used to measure the accumulated distance and directionality using the Chemotaxis plugin available for Image J.
Saha P., Yeoh B.S., Olvera R.A., Xiao X., Singh V., Awasthi D., Subramanian B.C., Chen Q., Dikshit M., Wang Y., Parent C.A, & Vijay-Kumar M. (2017). Bacterial Siderophores Hijack Neutrophil Functions. Journal of immunology (Baltimore, Md. : 1950), 198(11), 4293-4303.
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6

Imaging Yersinia Infection in BMDMs

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BMDMs seeded onto glass coverslips were left uninfected or infected as described above, except that tissue culture medium was supplemented with 0.5 mM IPTG to induce GFP expression in Y. pseudotuberculosis strains harboring p67GFP3.1. BMDMs were washed, fixed, permeablized and blocked, as previously described (Pujol and Bliska, 2003 (link)), and incubated with rabbit anti-Yersinia (Black and Bliska, 2000 (link)) or rat monoclonal anti-ASC (a gift from Gabriel Nuñez) antibodies. Binding of primary antibodies was visualized using FITC-conjugated anti-rabbit antibody (Sigma Immunochemicals) or Alexa Fluor 594-conjugated anti-rat antibody (Invitrogen) and DNA was stained with DAPI. Coverslips were mounted onto glass slides and imaged with an Axiovert S100 microscope (Zeiss). Three random fields of approximately 50 cells each were taken with a SPOT camera (Diagnostic Instruments) and processed with Adobe Photoshop.
Chung L.K., Park Y.H., Zheng Y., Brodsky I.E., Hearing P., Kastner D.L., Chae J.J, & Bliska J.B. (2016). The Yersinia Virulence Factor YopM Hijacks Host Kinases to Inhibit Type III Effector-Triggered Activation of the Pyrin Inflammasome. Cell host & microbe, 20(3), 296-306.
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7

Measurement of Ca2+ Dynamics in SH-SY5Y Cells

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Ca2+ measurements were performed essentially as described previously [20 (link), 78 (link), 79 (link)]. SH-SY5Y cells were loaded with 2 μM Fluo4-AM and/or Rhod2-AM dye (Invitrogen) in external solution (145 mM NaCl, 2 mM KCl, 5 mM NaHCO3, 1 mM MgCl2, 2.5 mM CaCl2, 10 mM glucose, 10 mM Na-HEPES, pH 7.25) containing 0.02% Pluronic-F27 (Invitrogen) for 15 min at 37 °C, followed by washing in external solution for 15 min at 37 °C. Fluo4 and Rhod2 fluorescence were timelapse recorded (1 s intervals) at 37 °C using either an Axiovert S100 microscope (Zeiss) driven by MetaMorph (Molecular Dynamics) and equipped with GFP (Fluo4) and DsRed (Rhod2) filtersets (Chroma Technology), a 40× Plan-Neofluar 1.3NA objective (Zeiss), and a Photometrics Cascade-II 512B36 EMCCD camera or a Nikon Ti-E microscope using a CFI Plan Apo VC 20× objective and Nikon Andor Neo sCMOD high-resolution camera and appropriate filter sets. The cells were kept under constant perfusion with external solution (0.5 ml/min). Inositol 1,4,5-trisphosphate (IP3) receptor-mediated Ca2+ release from ER stores was triggered by application of 100 μM Oxotremorine-M for 2 min. Ca2+ levels were calculated as relative Rhod2 or Fluo4 fluorescence compared to baseline fluorescence (F/F0) at the start of the measurement. Oxotremorine-M was from Santa Cruz Biotechnology and was dissolved in water.
Paillusson S., Gomez-Suaga P., Stoica R., Little D., Gissen P., Devine M.J., Noble W., Hanger D.P, & Miller C.C. (2017). α-Synuclein binds to the ER–mitochondria tethering protein VAPB to disrupt Ca2+ homeostasis and mitochondrial ATP production. Acta Neuropathologica, 134(1), 129-149.
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8

Cell Cycle Analysis of sEV-Exposed B16F1 Cells

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Changes in the cell cycle dynamics of sEV-exposed B16F1 cells were analysed using the Cell-Clock cell cycle assay (Biocolor Ltd) according to the assay protocol. This assay can be used to distinguish the four major phases of the mammalian cell cycle using a vital redox dye, which is yellow, green or dark blue in G1, S/G2, and M phase cells, respectively. After staining, cells were photographed using an Axiovert S100 microscope (Zeiss) equipped by a Nikon D5000 camera. Images were analysed by the ImageJ software to determine the percentage of cells in each cell cycle phase. The experiment was performed with 4 repeats.
Harmati M., Gyukity-Sebestyen E., Dobra G., Janovak L., Dekany I., Saydam O., Hunyadi-Gulyas E., Nagy I., Farkas A., Pankotai T., Ujfaludi Z., Horvath P., Piccinini F., Kovacs M., Biro T, & Buzas K. (2019). Small extracellular vesicles convey the stress-induced adaptive responses of melanoma cells. Scientific Reports, 9, 15329.
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9

Culturing B16F1 Mouse Melanoma Cells

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In this study, we used the B16F1 mouse melanoma cell line, which was obtained from ECACC (Salisbury, UK) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Lonza, Basel, Switzerland) supplemented with 10% FBS (EuroClone, Pero, Italy; or Biowest, Nuaillé, France), 2 mM L-glutamine, 1% Penicillin–Streptomycin–Amphotericin B mixture (P/S/A), 1% MEM non-essential amino acids, 1% MEM vitamin solution and 0.01% sodium pyruvate (all from Lonza, Basel, Switzerland). The B16F1 cell culture was maintained in a humidified incubator at 37 °C and 5% CO2. During the experiments, the morphology of the cells was monitored daily and the cells were stained with trypan blue dye (Corning, NY, USA) to count live and dead cells in a Bürker chamber to monitor cell viability. To compare different cultures, the same number of cells were seeded. The phase contrast microscopy images were made using an Axiovert S100 microscope (Zeiss, Oberkochen, Germany) at 20× magnification.
Böröczky T., Dobra G., Bukva M., Gyukity-Sebestyén E., Hunyadi-Gulyás É., Darula Z., Horváth P., Buzás K, & Harmati M. (2023). Impact of Experimental Conditions on Extracellular Vesicles’ Proteome: A Comparative Study. Life, 13(1), 206.
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10

Imaging Immune Response to GAS Infection

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C57Bl/6 female mice (n = 3) were infected intra-dermally with 5×107 M18 GAS and whole mount tissue sections and draining lymph nodes imaged 6 hours post infection. Whole mount tissue staining was carried out as described previously [2 (link)]. Frozen sections of lymph node were prepared by cryostat and fixed for 5 minutes in ice-cold acetone. Samples were stained with polyclonal anti-mouse LYVE-1 Ab and FITC-conjugated anti-group A carbohydrate antibody (Abcam). Whole mount tissue samples were viewed by Zeiss LSM780 confocal microscope, using either a Plan-Apochromat 10x/0.3DIC M27 (total magnification: 100x) or Plan-Apochromat 63x/1.4 oil (total magnification: 630x, resolution: 0.24 μm). For frozen lymph node sections, nuclei were counterstained with DAPI and sections were viewed on a Zeiss Axiovert S100 microscope equipped with epi-fluorescence.
Lynskey N.N., Banerji S., Johnson L.A., Holder K.A., Reglinski M., Wing P.A., Rigby D., Jackson D.G, & Sriskandan S. (2015). Rapid Lymphatic Dissemination of Encapsulated Group A Streptococci via Lymphatic Vessel Endothelial Receptor-1 Interaction. PLoS Pathogens, 11(9), e1005137.
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