Live/Dead Double Staining Kit (Merck)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Prior to use, mix 1 µl of solution A and 1 µl of solution B in 1ml of Staining Buffer	Resuspend to 1 ml Staining Solution. Incubate for 15 minutes at 37°C

Upstream tips
Prior to use, mix 1 µl of solution A and 1 µl of solution B in 1ml of Staining Buffer
Protocol tips
Resuspend to 1 ml Staining Solution. Incubate for 15 minutes at 37°C

Publication protocol

Live/dead bacterial double staining was used to distinguish dead bacteria (red staining) from live bacteria (green staining) using laser confocal microscopy (LSCM) (Fv-1000, Olympus, Japan). First, 2 mL of primary inoculum were inoculated on the membranes in a 12-well plate. After a 24 h culturing period, the membranes were taken out of the plates, washed with PBS three times, placed on new plates, and then stained using the Live/dead staining kit. Subsequently, several decimal solutions were sampled from the 12-well plate after the exposure of bacteria to the CCM membranes. Then, 30 μL of the diluted bacterial solution was spread on the BHI agar plates and incubated at 37 °C. After 24 h, the numbers of the surviving colonies were counted. Then bacteriostatic rate can be obtained from the following equation:

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Manufacturer protocol

Download the product protocol from Merck Millipore for Live/Dead Double Staining Kit (Merck) below.

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