RealTime-Glo assay
(Promega) was used since it allows continuous monitoring of the metabolic
activity of mammalian or bacterial cells and has particularly good
sensitivity for Gram-positive bacteria.17 (link) Bacterial suspensions (1 mL) were mixed with 1 μL of MT cell
viability substrate and 1 μL of NanoLuc enzyme, and incubated
in the dark for 1 h at 37 °C and 220 RPM. Bacterial suspensions
(40 μL) were then applied to disks within a white, opaque 24-well
plate, which was then sealed with transparent film (Greiner Bio-one
EasySeal plate sealer) to ensure sterility and to prevent the surfaces
from drying out. The plate was placed in a preheated (37 °C)
plate reader (Tecan infinite F200 Pro) and luminescence recorded every
10 min for up to 18 h with 1000 ms integration time, wait time of
0.1 s, and settle time of 150 ms.
(Promega) was used since it allows continuous monitoring of the metabolic
activity of mammalian or bacterial cells and has particularly good
sensitivity for Gram-positive bacteria.17 (link) Bacterial suspensions (1 mL) were mixed with 1 μL of MT cell
viability substrate and 1 μL of NanoLuc enzyme, and incubated
in the dark for 1 h at 37 °C and 220 RPM. Bacterial suspensions
(40 μL) were then applied to disks within a white, opaque 24-well
plate, which was then sealed with transparent film (Greiner Bio-one
EasySeal plate sealer) to ensure sterility and to prevent the surfaces
from drying out. The plate was placed in a preheated (37 °C)
plate reader (Tecan infinite F200 Pro) and luminescence recorded every
10 min for up to 18 h with 1000 ms integration time, wait time of
0.1 s, and settle time of 150 ms.
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