SYTO™ 9 Green Fluorescent Nucleic Acid Stain

Live / Dead assay bacteria - Pseudomonas aeruginosa

Experiment

Live / Dead assay bacteria - Pseudomonas aeruginosa

Product

SYTO™ 9 Green Fluorescent Nucleic Acid Stain from Thermo Fisher Scientific

Manufacturer

Thermo Fisher Scientific

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 50 μl of 2.5 μM SYTO9 to the wells. Incubate on the orbital shaker for 15 minutes in the dark.	Detect using a 488/20 nm excitation filter (for both SYTO9 and PI) and a 528/20 nm (SYTO9 emission wavelength) and 645/40 nm (PI emission wavelength) emission filter.

Protocol tips
Add 50 μl of 2.5 μM SYTO9 to the wells. Incubate on the orbital shaker for 15 minutes in the dark.
Downstream tips
Detect using a 488/20 nm excitation filter (for both SYTO9 and PI) and a 528/20 nm (SYTO9 emission wavelength) and 645/40 nm (PI emission wavelength) emission filter.

Publication protocol

100 μl of cell suspension were added per well of a black 96-well plate (BRANDplates® pureGrade™). 50 μl of 2.5 μM SYTO9 (S-34854, Molecular Probes®) was added per well before incubating on the orbital shaker for 15 minutes in the dark. Fluorescence intensity was measured with a Synergy HT Multi-Detection Microplate Reader (BioTek®) using a 488/20 nm excitation filter (for both SYTO9 and PI) and a 528/20 nm (SYTO9 emission wavelength) and 645/40 nm (PI emission wavelength) emission filter. 50 μl of 15 μM propidium iodide was added per well before incubating additional 15 minutes on the orbital shaker in the dark and measuring fluorescence intensity with the same filter sets.

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Discussion

5 years ago

Author: M. Daecher Germany

Live/dead assay Bacteria

Hello everyone! I am going to do a live/dead assay for my cells and I saw that I can use both fluorescence and absorbance as my detection method. Is there a difference in the results depending on the method? Is one method preferred over the other in certain situations?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Live / Dead assay bacteria - Pseudomonas aeruginosa using SYTO™ 9 Green Fluorescent Nucleic Acid Stain from Thermo Fisher Scientific.

Paper title

Critical aspects of using bacterial cell viability assays with the fluorophores SYTO9 and propidium iodide

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for SYTO™ 9 Green Fluorescent Nucleic Acid Stain below.

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