Resazurin viability assay

LS Lorgio Victor Bautista Samaniego
SS Samuel Luis Scandelau
CS Caroline Rosa Silva
SP Sebastião Pratavieira
VP Vanessa de Oliveira Arnoldi Pellegrini
AD Andrei Nicoli Gebieluca Dabul
LE Luís Antônio Esmerino
MN Mario de Oliveira Neto
RH Rodrigo Tavanelli Hernandes
FS Fernando Segato
MP Marcos Pileggi
IP Igor Polikarpov
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The percentage of metabolically active bacteria was determined by adding 20 µL of a 0.15 mg/mL resazurin (Sigma-Aldrich, St. Louis, USA) solution to 100 µL of the resuspended cells. This mixture was incubated at 37 °C for 2 h in a 96-well black plate in static conditions and the fluorescence was measured at the excitation/emission wavelengths of 550/590 nm, respectively, in an Infinite 200 M PRO microplate reader (Tecan, Hombrechtikon, Switzerland). The percentage of metabolically active cells was calculated using the Eq. (8):

where SC550/590 and GC550/590 correspond to the fluorescence measurements of the control without inoculation and the biofilm control without enzyme treatment, respectively. The experiments were performed in triplicate.

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