Please help to differentiate between intracellular particles and bacterial contamination.
Hi Vidya,
In a cell-culture contamination there is always change in pH which can be visible by the change in media colour. Since the bacterial cells grow at very high rate, this change in media colour is visible rather quickly. Another thing you can notice is the development of turbidity in the media. These things are visible when you have very contamination.
But since most of times we use some antibiotics in the media, therefore SOMETIMES contamination may not be visible without microscope. While observing your culture flask or plate under microscope at high resolution, if you find something having wiggling movements then its a bacterial contamination. The cellular debris or dead particles dont show this type of wiggly movements, they move at very slow pace because of the media disturbance occurred while handling the flask. If you want to see the difference in movement, you can use one drop of bacterial culture on a slide under microscope and see their fast movement.
The visualization of the mycoplasma and fungal contamination is not very easy with the microscopes, for that you need to have some chemical tests which are available commercially. You can check mycoplasma contamination using Hoechst dye. Protocol should be easily available on internet.
Something to further help you.
https://www.thermofisher.com/fi/en/home/references/gibco-cell-culture-basics/biological-contamination.html
Hope it will help you. All the best.
Bacteria are distinguished by having DNA in them. You can stain them with DAPI. Example images & protocols are easily found on the internet.
Hi Vindya,
Bacterial cells divide at a rapid rate and show increased counts over very short time spans. On the other hand, intracellular particles wont divide and grow. A rough estimate of counts (even of small areas) over time might give you an idea what they are actually.
Good luck!
There are some contaminants I have experienced before. Bacterial contamination obviuosly and rapidly changes the medium in 16
Hi Vidya,
In a cell-culture contamination there is always change in pH which can be visible by the change in media colour. Since the bacterial cells grow at very high rate, this change in media colour is visible rather quickly. Another thing you can notice is the development of turbidity in the media. These things are visible when you have very contamination.
But since most of times we use some antibiotics in the media, therefore SOMETIMES contamination may not be visible without microscope. While observing your culture flask or plate under microscope at high resolution, if you find something having wiggling movements then its a bacterial contamination. The cellular debris or dead particles dont show this type of wiggly movements, they move at very slow pace because of the media disturbance occurred while handling the flask. If you want to see the difference in movement, you can use one drop of bacterial culture on a slide under microscope and see their fast movement.
The visualization of the mycoplasma and fungal contamination is not very easy with the microscopes, for that you need to have some chemical tests which are available commercially. You can check mycoplasma contamination using Hoechst dye. Protocol should be easily available on internet.
Something to further help you.
https://www.thermofisher.com/fi/en/home/references/gibco-cell-culture-basics/biological-contamination.html
Hope it will help you. All the best.
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