Cells were fixed with 4% paraformaldehyde in PBS for 20 min at room temperature and washed with PBS. Cells were permeabilized with 0.1% Triton X-100 in PBS at room temperature for 5 min. After blocking with 3% non-fat dry milk for 30 min, cells were incubated with mouse monoclonal anti-influenza A nucleoprotein (AbD Serotec); bound antibodies were revealed with goat anti-mouse IgG conjugated with Alexa Fluor 568 (Molecular Probes). After washing, nuclei were stained with 1 μg ml−1 4′,6-diamidino-2-phenylindole (DAPI; Molecular Probes) in PBS for 15 min at room temperature. Fluorescent images were acquired on an Olympus IX70 microscope equipped with Nanomover and softWoRx DeltaVision image acquisition software (Applied Precision, WA, USA) and a U-PLAN-APO 60× objective. Images were captured under constant exposure time, gain and offset. To improve the contrast and resolution of digital images captured in the microscope, they were elaborated with deconvolution software.
Nanomover and softworx deltavision image acquisition software
Manufactured by Cytiva
Sourced in United States
The Nanomover is a high-precision motorized stage designed for microscopy applications. It provides accurate and repeatable positioning of samples during image acquisition. The softWoRx DeltaVision image acquisition software is a comprehensive solution for controlling DeltaVision microscope systems and processing the resulting image data.
Automatically generated - may contain errors
2 protocols using nanomover and softworx deltavision image acquisition software
1
Immunofluorescence Staining of Influenza A
2
Immunofluorescent Detection of HCV Core
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Cells were fixed in methanol : acetone (50 : 50, V/V) for 20 min at -20°C. After blocking with PBS containing 1% bovine serum albumin (BSA) for at least 30 min, cells were incubated with mouse monoclonal antihepatitis C virus core 1b antibody (Abcam, AB 58713); bound antibodies were revealed with mouse anti-human IgG conjugated with Alexa Fluor 488 (Life Technologies, Z25102). After washing, the nuclei were stained with 1 μg/ml 4,6-diamidino-2-phenylindole (DAPI; Molecular Probes) in PBS for 15 min at room temperature. Fluorescent images were acquired on an Olympus IX70 microscope equipped with Nanomover and softWoRx DeltaVision image acquisition software (Applied Precision, WA, USA) and a U-PLAN-APO 40× objective. Images were captured under constant exposure time, gain, and offset.
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