Control- and ISG15-siRNA transfected HTR8/SVneo cells seeded onto 8-well culture slides (BD biosciences, Bedford, MA, United States) and were fixed with 4% paraformaldehyde for 20 min at 4°C. After washing and permeabilization steps, the slides were incubated with TRITC-conjugated phalloidin (Millipore, San Diego, CA, United States), which preferentially binds to actin filaments, for 30 min at room temperature. After washing, the slides were treated with 6-diamino-2-phenylindole (DAPI) for 10 min and mounted with antifade mounting medium (Vector Lab). Slides were examined under Axio Observer-Z1 immunofluorescence/phase-contrast microscope (Zeiss) with ZEN Imaging system (Zeiss).
Zen imaging system
Manufactured by Zeiss
Sourced in Germany
The Zen imaging system is a software platform developed by Zeiss for the acquisition, processing, and analysis of microscopy images. It provides a user-friendly interface to control and configure various Zeiss microscopes and cameras, enabling researchers to capture high-quality images and perform essential image analysis tasks.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Antifade mounting medium, by Vector Laboratories (1 mentions)
Tritc conjugated phalloidin, by Merck Group (1 mentions)
8 well culture slide, by BD (1 mentions)
Sc 271430, by Santa Cruz Biotechnology (1 mentions)
Dapi g1012, by Wuhan Servicebio Technology (1 mentions)
4 protocols using zen imaging system
1
Visualizing Actin Cytoskeleton in HTR8/SVneo Cells
2
Immunohistochemical Profiling of Retinal ATP8A Proteins
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Cryosections of adult mouse retinal tissues were fixed with 4% paraformaldehyde in 100 mM phosphate buffer (PB, pH 7.4) for 60 min as previously described54 (link). The sections were subsequently blocked, and permeabilized with PB containing 0.2% Triton X-100 and 10% normal goat serum for 30 min and labeled overnight at room temperature with antibodies to ATP8A2 and ATP8A1 diluted 1:200 in PB containing 2.5% normal goat serum and 0.1% Triton X-100. Sections were washed 3 times with PB and labeled for 3 h with goat anti-rabbit Alexa-594 secondary antibody (diluted 1:1000) and counterstained with DAPI nuclear stain. The sections were examined on a Zeiss LSM 700 confocal microscope with a Zen imaging system.
3
Immunohistochemistry of Microglia Markers
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Brains sections were immersed in 0.1% Triton X-100 for 10 min, blocked for 1 h, and then exposed to primary antibodies at 4°C overnight, including rabbit anti-Iba1 (019-19741, Wako, Chuo-Ku, Osaka, Japan) (1 : 500) and mouse anti-CD68 (MCA341R, Bio-Rad) (1 : 50) or mouse anti-Arg-1 (sc-271430, Santa Cruz Biotechnology, Dallas, TX, USA) (1 : 25). Subsequently, the incubation solution was replaced by goat anti-rabbit IgG (A-21428, Thermo Fisher Scientific) (1 : 1000) and -mouse IgG (A-11029, Thermo Fisher Scientific) (1 : 500). Finally, sections were counterstained with DAPI (G1012, Servicebio, Wuhan). Images were recorded using a ZEN imaging system (Zeiss, Jena, Germany) at ×200 magnification.
4
Placental Explant Immunofluorescence Protocol
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Cultured placental explants were fixed in 4% paraformaldehyde for 2h, then transferred to phosphate-buffered saline (PBS) for 24h. Samples were embedded in paraffin or in OCT (for QD localisation), sectioned (5µm), and mounted on glass slides. Paraffin embedded sections were dewaxed in xylene and rehydrated. Antigen retrieval was performed by boiling in 0.1 M sodium citrate buffer, pH 6.3 for 10 min or by incubating in 2mM EDTA pH 6.0 at 80°C for 20 min (Table 1). Tissues were treated with sodium borohydride (10mg/ml, 3x10min), followed with a PBS wash, to decrease syncytial autofluorescence, then incubated with primary antibody overnight at 4°C and with fluorescent secondary antibody for 1h at room temperature (Table 1). Images were taken using the Zen imaging system (Zeiss). Control experiments included omission of primary or secondary antibodies, or substitution of primary antibody with mouse or rabbit IgG.
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