Antibacterial activity determination with microdilution method without. Microplate reader, i need a available stain or pigment which give me a colour indicates the viability of bacteria
The standard method is simpler and more reliable: You measure the turbidity of the culture, either by eye or by using an absorbance plate reader set at 600 nm, after allowing the uninhibited culture to grow to stationary phase (turbid) overnight. The minimal inhibitory concentration (MIC) is in the clear well with the lowest compound concentration.
The standard method is simpler and more reliable: You measure the turbidity of the culture, either by eye or by using an absorbance plate reader set at 600 nm, after allowing the uninhibited culture to grow to stationary phase (turbid) overnight. The minimal inhibitory concentration (MIC) is in the clear well with the lowest compound concentration.
if you have a limited number of samples you can growth bacteria in larger volumes and check the od600 with standard single cell cuvette uv-vis reader. Of course if you have many samples or your antimicrobial molecule is expensive microplate are better. 2 alternatives to the od600 may be the the xtt assay or cell titer blue that are viability assay characterized from the secretion of coloured compounds by the viable cells and expecially cell titer blue where you have strong agift from blue to pink may allow you to perform an estimation aquiring a picture of the plate and performing an image analisys by a software as imagej or similar. however this probes are more sensitive to live bacteria than od600 therefore you have to reduce the inocula of bacteria incubated with the antimicrobial peptides under their detection limit otherwise in this way you will determine the mbc but not the mic.
good luck
Manuele
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining