Three strains were selected for the SEM, Candida krusei (ATCC 6258), C. glabrata (ATCC 2001) and C. parapsilosis 50. Yeast inoculum was adjusted according to the bio lm formation assay. Inoculum suspension and cell adhesion procedures were performed in a 24 well polystyrene plate (JET BIOFIL®, China) and washed as described above. Pre-formed bio lms were incubated for 24h at 37°C with 100 µl of YNB medium, along with the concentrations used in the antibio lm activity assays. Concomitantly, the mature bio lm was incubated for 24h at 37°C, with 100 µl of YNB medium. Then, the plate was treated with 1.25 mg/ml and 2.5 mg/ml of Ag2 fraction and incubated for 24h at 37°C. The bio lms were xed with 2% (v/v) glutaraldehyde (Sigma) and dehydrated with an ethanol solution series (50%, 70%, 90% and 100%). Samples were taken, dried, glued on microscopy slides and metallized with 24k gold and then subjected to visualization on the SEM apparatus (JEOL, JSM 5600LV, Japan). An in formation and mature bio lms not exposed to the Ag2 fraction were used as control. Treated and untreated bio lm samples followed the protocol described by Barbosa and colleagues [25] .
24 well polystyrene plate
Manufactured by Jet Biofil
Sourced in China
The 24 well polystyrene plate is a laboratory equipment designed for cell culture and assay applications. It provides a standardized and uniform format for conducting experiments involving multiple samples or conditions. The product is made of polystyrene, a commonly used material in the field of biotechnology.
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2 protocols using 24 well polystyrene plate
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Candida Biofilm Disruption by Silver Fraction
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Candida Biofilm Disruption by Silver Fraction
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Three strains were selected for the SEM, Candida krusei (ATCC 6258), C. glabrata (ATCC 2001) and C. parapsilosis 50. Yeast inoculum was adjusted according to the bio lm formation assay. Inoculum suspension and cell adhesion procedures were performed in a 24 well polystyrene plate (JET BIOFIL®, China) and washed as described above. Pre-formed bio lms were incubated for 24h at 37°C with 100 µl of YNB medium, along with the concentrations used in the antibio lm activity assays. Concomitantly, the mature bio lm was incubated for 24h at 37°C, with 100 µl of YNB medium. Then, the plate was treated with 1.25 mg/ml and 2.5 mg/ml of Ag2 fraction and incubated for 24h at 37°C. The bio lms were xed with 2% (v/v) glutaraldehyde (Sigma) and dehydrated with an ethanol solution series (50%, 70%, 90% and 100%). Samples were taken, dried, glued on microscopy slides and metallized with 24k gold and then subjected to visualization on the SEM apparatus (JEOL, JSM 5600LV, Japan). An in formation and mature bio lms not exposed to the Ag2 fraction were used as control. Treated and untreated bio lm samples followed the protocol described by Barbosa and colleagues [25] .
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