HeLa and A549 cells were treated with CDDP (1.25 µg/mL) at the indicated time (4, 8, 12, and 16 h). After fixation, blocking and permeabilization, the cells were incubated with the anti-HuR primary antibody, followed by Alexa Fluor 488 secondary antibodies. Cell nuclei were stained with DAPI before mounted on slides by using Mountant permafluor (Thermo scientific, FM 111212A). Cells were observed using an IX71 inverted microscope, and image acquisition was performed with the Olympus Fluoview Software (FV10-ASW 4.2 viewer).
Fluoview software fv10 asw 4.2 viewer
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The Fluoview Software (FV10-ASW 4.2 viewer) is a comprehensive imaging software designed for use with Olympus confocal microscope systems. It provides a user-friendly interface for capturing, processing, and analyzing fluorescence images. The software supports a range of imaging techniques, including confocal, two-photon, and multi-dimensional imaging.
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3 protocols using fluoview software fv10 asw 4.2 viewer
1
CDDP-Induced HuR Localization Dynamics
2
Immunofluorescence Staining of HuR in Cells
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A549 and HeLa cells were grown over coverslips in 24 well plates at 60–80% confluence and treated with LPS (25 ng/mL). For HuR staining, cells were rinsed twice with phosphate buffered saline (PBS), fixed for 10 min at RT with 4% paraformaldehyde in PBS at room temperature, blocking and permeabilization was completed using 1% BSA plus 0.25% Triton X-100 in PBS at room temperature. The fixed cells were subsequently incubated with HuR primary antibody for overnight at 4 °C followed by Alexa Fluor 488 secondary antibodies for 1 h at room temperature, respectively. DAPI was used to counterstain cell nuclei before cells were mounted on slides using Mountant permafluor (Thermo scientific, FM 111212A, Waltham, MA, USA). Cells were observed with an IX71 inverted microscope (Olympus, Tokyo, Japan). Image acquisition was performed with the Olympus Fluoview Software (FV10-ASW 4.2 viewer).
3
Visualizing Actin Cytoskeleton Dynamics
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Cells were grown on coverslips in 35-mmdishes at 60-80% confluence and were stimulated for 2 h with Angin II without or with microfilament inhibitor either latrunculin A (Lat.A;30 min) or blebbistatin (Blebbistat;4 hour). Immediately following treatment, cells were rinsed once with phosphate buffered saline (PBS), fixed for 20 min at RT with 4% paraformaldehyde in PBS, blocked and permeabilized in 2% BSA plus 0.1% Triton X-100 (PBS) at room temperature (RT). Subsequently, the fixed cells were incubated with anti-HuR and myosin II primary antibody for overnight at 4ºC followed by Alexa Fluor 488 and Alexa Fluor 568 secondary antibodies for 1 hour at room temperature respectively. F-actin was visualized with rhodamine-conjugated phalloidin.
Cell nuclei were counterstained with DAPI before mounted on slides by using Mountant permafluor (Thermo scientific, FM 111212A). Cells were observed using an IX71 inverted microscope (Olympus). Image acquisition was performed with the Olympus Fluoview Software (FV10-ASW 4.2 viewer).
Cell nuclei were counterstained with DAPI before mounted on slides by using Mountant permafluor (Thermo scientific, FM 111212A). Cells were observed using an IX71 inverted microscope (Olympus). Image acquisition was performed with the Olympus Fluoview Software (FV10-ASW 4.2 viewer).
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