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Sections were counterstained with hematoxylin and mounted for microscopy.
The podocytes then were fixed, blocked and stained with FITC-labeled phalloidin and DAPI and mounted for microscopy.
Following washing 3X in PBS the coverslips were mounted for microscopy in Vectashield hard-set mounting medium (Vector Laboratories, Burlingame, CA).
After immnuostaining, some sections were treated with 0.3% 4',6'-diamidino-2-phenylindole (DAPI; Sigma, St Louis, MO) for 15 min, washed and mounted for microscopy.
Phenotypic characterization of Mn treated worms.- Following Mn treatment up to 24 h post-treatment, worms were observed under stereomicroscope (Zeiss), and mounted for microscopy phenotypic analysis using our Nikon platform aforementioned.
Cells grown on cover glass were stained with 10 ng/ml Hoechst33342 (Sigma) for 30 min, and fixed in 2% formaldehyde for 10 min at room temperature, washed with phosphate-buffered saline, and mounted for microscopy.
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These fixed biofilms were rinsed 5 times with 0.9% saline before mounting for microscopy.
The slides were mounted for light microscopy with Clarion™ Mounting Medium (Sigma-Aldrich, St . Louis USA).
The nuclei were counterstained with 10 μg/ml DAPI (Life Technologies™, New York, USA) at room temperature for 10 min, and the slides were mounted for confocal microscopy with anti-quenching fluoromount (Sigma-Aldrich, St . Louis USA).
Finally, samples were mounted for fluorescence microscopy by using ProLong antifade kit (Invitrogen).
Finally, cells were washed and mounted for flurescence microscopy by using ProLong antifade kit (Invitrogen).
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