LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy

Protocol tips

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	Ad Syto-9 and propidium iodide (PI) at a concentration of 0.01 mM and 0.06 mM,respectively.

Protocol tips
Ad Syto-9 and propidium iodide (PI) at a concentration of 0.01 mM and 0.06 mM,respectively.

Publication protocol

The biofilms were examined by Zeiss LSM 510 META equipped with a water-immersion of 63×objective (Carl Zeiss, Jena, Germany). The biofilms were stained for 15 min with 100 µl LIVE/DEAD BacLight Bacterial Viability Kit (Invitrogen Corporation, NY, USA). The final concentrations of Syto-9 and propidium iodide (PI) were 0.01 mM and 0.06 mM, respectively. The SYPRO® Ruby biofilm matrix stain (Invitrogen Corporation, NY, USA) was used to stain proteins in the EPM. The green fluorescence and red fluorescence of SYTO 9 and PI were excited using an argon laser beam, with excitation lines at 488 nm and a helium/neon at 543 nm, respectively. The SYPRO® Ruby stain was excited at 405 nm with diode laser. The CLSM image stacks were analysed by the image-processing software COMSTAT (28). The biomass, average thickness, and maximum thickness were the parameters used to compare different biofilms.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy below.

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