To visualize the viable and dead cells, control cells and cells treated with G-AgNPs were stained for 20 min with a mixture of 6.0 μM SYTO 9 and 30 μM KI from Live/Dead BacLight Viability kit L13152 (Invitrogen, Molecular Probes, Inc., Eugene, OR, United States). Fluorescence microscopic imaging of the cells was performed using a LEICA DM 4000 B (Leica Microsystems, Copenhagen, Denmark).
Live dead baclight viability kit l13152
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Live/Dead BacLight Viability kit L13152 is a fluorescence-based assay used to determine the viability of bacterial cells. The kit contains two nucleic acid stains, SYTO 9 and propidium iodide, which differentially label live and dead cells. The assay enables the quantification of viable and non-viable bacterial populations through fluorescence microscopy or flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using live dead baclight viability kit l13152
1
Cell Viability Staining with SYTO 9 and KI
2
Quantifying Live and Dead Cells in Biofilms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The biofilms were stained using the Live/Dead BacLight Viability kit L13152, (Invitrogen, Molecular Probes, Inc. Eugene, OR, USA) to determine the proportion of live and dead cells. Bacterial cells with intact cell membranes emit green fluorescence while dead or damaged bacterial cells emit red fluorescence. The staining was performed at room temperature in the dark, for 20 min, using a mixture of 6.0 μM SYTO 9 and 30 μM potassium iodide. Fluorescence microscopic imaging of the biofilms was performed using a Zeiss fluorescence microscope (Axio Imager.Z2m Carl Zeiss, Zena, Germany).
3
Live/Dead Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To visualize the viable and dead cells, we used the Live/Dead BacLight Viability kit L13152 (Invitrogen, Molecular Probes, Inc., Eugene, OR, USA). Control and treated cells were stained for 20 min with a 6.0 μM SYTO 9 and 30 μM KI mixture. Fluorescence microscopic imaging of the cells was performed using a LEICA DM 4000 B (Leica Microsystems, Denmark) [20 (link)].
4
Biofilm Inhibition by Silver Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three different methods analyzed the biofilm inhibition activity of AgNPs: (1) by checking the bacterial viability, (2) by observing the cells under the scanning electron microscope (SEM), and (3) by live/dead staining assay25 (link),28 (link). The inoculum of 2–5 × 106 CFU/mL was obtained by diluting overnight cultures of E. coli and P. aeruginosa and loading the samples on top of a 15 mm cover glass. The cover glass was further incubated at 37 °C for 24 h, which allowed for biofilm formation. After 24 h, old culture medium was replaced with fresh medium, containing sterile water (negative control) or 1 × MBC, 2 × MBC, 4 × MBC, or 8 × MBC of AgNPs. Sample were incubated for further 24 h. Then the biofilms were homogenized for 20 s in 0.89% NaCl. The obtained bacterial samples were serially diluted and plated on LB agar plates. Next, the plates were incubated overnight at 37 °C, followed by CFU counting. For live/dead cells staining, samples were stained for 20 min with a mixture of 6.0 μM SYTO 9 and 30 μM KI from Live/Dead BacLight Viability kit L13152, (Invitrogen, Molecular Probes, Inc. Eugene, Oregon, USA). Fluorescence microscopy were collected using a Zeiss fluorescence microscope (Axio Imager.Z2m Carl Zeiss, Zena, Germany). Further, control and AgNPs-treated biofilms were examined by SEM (Supra 60 VP microscope, Carl Zeiss AG)28 (link).
5
Silver Nanoparticles Inhibit Bacterial Adhesion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AgNPs were coated on 15 mm glass cover slips by drop casting method [29] . Briefly, 250 ml of samples with different concentrations of water dispersed nanoparticles were placed on cover glass and dried at 37 C overnight. The coated cover glass slides were used to evaluate the effect of these nanoparticles on bacterial adhesion. 250 ml of bacterial suspension containing 1-2 Â 10 6 CFUs/ml were added to the non-coated and coated cover glass and incubated for 5 h at 37 C. After incubation, biofilms were rinsed with sterile water and homogenized by sonication. The homogenized biofilm suspensions were plated on agar plates for CFU counting. Furthermore, adhesion of the bacterial cells on the AgNPs coated and noncoated surface was visualized by using fluorescence microscopy as described previously [30] . Briefly, after a 5 h period of bacterial adhesion, biofilms were rinsed with sterile water and stained with the mixture of 6.0 lM SYTO 9 and 30 lM potassium iodide from Live/Dead BacLight Viability kit L13152 (Invitrogen, Molecular Probes, Inc. Eugene, OR) for 20 min. The imaging of stained biofilms was performed by using a fluorescence microscope (Axio Imager.Z2m Carl Zeiss, Zena, Germany).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!