Primary hepatocytes from TAK1FL/FL and TAK1ΔHEP mice were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. After washing with PBS, the hepatocytes were incubated with BODIPY™ 581/591 C11 (Invitrogen™ D3861, 5 μmol/L) for 30 min at 37 °C. DAPI was used to stain cell nuclei. Images were obtained on a Zeiss AxioObserver Z1 confocal microscope.
Axio observer z1 confocal microscope
Manufactured by Zeiss
Sourced in Germany
The Axio Observer Z1 is a confocal microscope produced by Zeiss. It is designed for high-resolution imaging and analysis of samples. The microscope utilizes laser scanning technology to capture detailed images of specimens.
Automatically generated - may contain errors
Lab products found in correlation
Fluoroshield medium, by Merck Group (3 mentions)
Paraformaldehyde (pfa), by Merck Group (3 mentions)
Rhodamine phalloidin, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Cellmask deep red, by Thermo Fisher Scientific (2 mentions)
Bodipy 581 591 c11, by Thermo Fisher Scientific (1 mentions)
Axio scan z1, by Zeiss (1 mentions)
Zen microscopy software, by Zeiss (1 mentions)
Rneasy plus micro kit, by Qiagen (1 mentions)
Palm microbeam system, by Zeiss (1 mentions)
Gdna eliminator column, by Qiagen (1 mentions)
Palm membrane slides, by Zeiss (1 mentions)
Rnasezap, by Thermo Fisher Scientific (1 mentions)
Palmrobo software 4, by Zeiss (1 mentions)
Quantitect whole transcriptome amplification kit, by Qiagen (1 mentions)
Lsm 710, by Zeiss (1 mentions)
16 protocols using axio observer z1 confocal microscope
1
Oxidative Stress Evaluation in Hepatocytes
2
Visualizing Cryptococcus neoformans Capsule in Brain Tissue
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C. neoformans cells and CPS released in tissue were stained using GXM-specific mAb 18B7. Slides were blocked, and mAb 18B7 (2 μg/ml) was added for 1 h at 37°C. After the slides were washed, fluorescein isothiocyanate (FITC)-conjugated GAM Ab [dilution, 1:250; 1% BSA] was applied for 1 h at RT. Neurons in tissue sections were stained with DAPI (4′,6-diamidino-2-phenylindole) and microtubule-associated protein-2 (MAP-2) as described previously [74 (link)]. Microscopic examinations of brain sections were performed with a fully motorized Carl Zeiss Axio Observer Z1 confocal microscope. Confocal images of blue, green, and red fluorescence were conceived simultaneously using a multichannel mode. Z-stack images and measurements were corrected utilizing Zen software in deconvolution mode.
3
Live Cell Imaging of EV Uptake
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For live cell imaging, RAS3 cells were treated with 1 × 1010 particles/ml of PKH26‐labeled EV preparation for 18‐h. Rapid time‐lapse imaging was performed using Zeiss spinning disk Axio Observer Z1 confocal microscope at 37°C within 5% CO2 environment. Live cell images were acquired every 5‐s using an Evolve 512 EMCCD camera with Alpha Plan‐Apo 63 × /1.46 objective. Cells were imaged for up to 5 min and particle tracking was performed using TrackMate for Fiji (Vesicle diameter = 1μm; thresholds and other parameters were chosen as appropriate based on control samples within each experiment).
4
Visualizing GPCR signaling mechanisms
5
Immunofluorescence Imaging of HeLa Cells
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HeLa cells were seeded on glass coverslips and were either mock-infected of infected at 10 PFU/cell or 2 PFU/cell in case of 16 h time point. At the indicated times, the cells were washed twice with PBS and fixed with 4% paraformaldehyde in PBS containing 250 mM HEPES. The cells were permeabilized with 0.1% triton X-100 followed by blocking with 10% FBS in PBS (blocking buffer) for 0.5 h. Coverslips were incubated with primary antibodies for 1 h in a moist chamber followed by three 10 min washes with 10% FBS. Coverslips were incubated with secondary antibody (Alexa Fluor 488 Goat Anti-Rabbit IgG (H+L), Alexa Fluor 546 donkey Anti-Mouse IgG (H+L) 1:750 diluted in blocking buffer) for 30 min in a moist chamber followed by three 5 min washes with 10% FBS and PBS only. The coverslips were washed with water and mounted in Mowiol 4–88 containing DAPI. Coverslips were allowed to set and stored at 4°C. Cells were visualized by Axio observer Z1 confocal microscope (Zeiss) with a 63x oil objective.
6
Visualization of Actin Cytoskeleton in Cells
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Cells were washed three times with PBS. The cover slips were transferred onto parafilm, fixed with 3.7% paraformaldehyde (Sigma, St. Louis, MO, USA), and rinsed three times with PBS. For visualization of actin microfilaments, cells were stained with rhodamine phalloidin (Invitrogen, Waltham, MA, USA) for 15 min at 37 °C in an atmosphere of 5% CO2. The preparations were rinsed three times with PBS, and embedded in Fluoroshield medium (Sigma, St. Louis, MO, USA). Intensity of staining of preparations was estimated using an AxioObserver Z1 confocal microscope (Carl Zeiss, Jena, Germany). In each experiment, at least 30 cells were imaged. Images were processed using ImageJ software [32 (link)].
7
Laser-Guided Micropattern Inscription on Azopolymer Surfaces
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ACS52telo were cultivated on the fibronectin micropatterned pDR1m coated glass slides for 24 h. Afterwards, real-time pattern inscription on cell-populated samples was performed by using an Axio Observer Z1 confocal microscope (Zeiss). The isomerization of the azopolymer and its consequent mass transport were activated using an Argon laser at 488 nm and 0.44 mW of intensity (measured with a PM100D optical meter (Thorlabs) with a 10 × (N.A. 0.3) objective lens. Patterns were embossed on the pDR1m substrates by illuminating with the laser light in specifically drawn regions of interest (ROIs). Parallel arrays of linear 100 µm long ROIs, separated by 700 nm, were inscribed by placing the circular cell in the centre (Figure 1A ). The exposure time was 60 s.
8
Fluorescent Labeling of Antibodies and Cells
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Single-domain antibody and trastuzumab were directly labeled by sulfo-Cy5 or AF488 according to standard protocol [34 ]. Briefly, protein solutions in 0.1 M Na2HPO4 pH 8.2 were mixed with excess of fluorescent dye NHS ester and incubated for 2 h at +4 °C. The unreacted dye was removed by buffer exchange on a Sephadex G25 column pre-equilibrated with PBS. The labeling efficiency was analyzed by measuring absorbance at 280 nm and 646 nm (Cy5) or 495 nm (AF488). Two days prior to experiment, 2 × 105 cells were seeded onto Petri dishes with cover slips. K562 cells were transferred onto coverslips after staining. After incubation under growing conditions, cells were washed twice with PBS, stained for 30 min in PBS with 5 μg/mL labeled antibodies and Hoechst 33342, and rinsed with PBS. Images were obtained on AxioObserver Z1 confocal microscope (Carl Zeiss).
9
Actin Cytoskeleton Remodeling Assay
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Cells were seeded onto Petri dishes with cover slips at a density of 2 × 105 cells per dish and incubated for 24 h. After that, cells were treated with compounds 4d , 4e , 4f , 4h , and 4i (5 μg/mL) for 24 h. The medium was removed, and cells were fixed with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA), washed three times with PBS, and permeabilized with 0.3% Triton-X100 (Sigma-Aldrich, St. Louis, MO, USA). The cells were rinsed three times with PBS. Actin filaments (microfilaments) were stained at 37 °C for 15 min with rhodamine-phalloidin (Invitrogen, Carlsbad, CA, USA). The samples were rinsed three times with PBS, followed by embedding in Fluoroshield medium (Sigma-Aldrich, St. Louis, MO, USA). Cells were imaged using an Axio Observer Z1 confocal microscope (Carl Zeiss MicroImaging GmbH, Jena, Germany). In each experiment, at least 30 cells were imaged. Images were analyzed by a pathologist, blinded to the treatment mode used for each group, using ImageJ software.
10
Confocal Imaging of Hippocampal-Entorhinal Projections
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For all but retrograde rabies experiments, slices were imaged using an inverted Zeiss Axio Observer Z.1 Confocal Microscope, using (magnification/NA) 10x/0.3 air, 20x/0.8 air, and/or 40x/1.30 oil immersion objectives (Zeiss) and 405, 488, 594, and 647 nm lasers for excitation to acquire 1024 × 1024 pixel, 16-bit image stacks. Images were acquired every 5 μm (image stack step-size) for the 20x overview images to trace the hippocampal projections to different layers of entorhinal cortex and 1 μm for 40x images used for neuronal reconstructions. Tiled images were stitched using Zen Microscopy Software (Zeiss), and the tiled z-stacks were subsequently processed using Fiji (ImageJ).
For retrograde rabies virus tracing experiments, all processed tissue from each series was first scanned using a Zeiss Axio Scan.Z1 (ZEISS). Images were acquired with 20x objectives, single focus in the Z-plane. Slides were scanned using a quadruple filter (preset for DAPI, dl488, dl546 and dl647), using LEDs from a Calibri 2 source to excite each corresponding fluorophore.
For retrograde rabies virus tracing experiments, all processed tissue from each series was first scanned using a Zeiss Axio Scan.Z1 (ZEISS). Images were acquired with 20x objectives, single focus in the Z-plane. Slides were scanned using a quadruple filter (preset for DAPI, dl488, dl546 and dl647), using LEDs from a Calibri 2 source to excite each corresponding fluorophore.
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