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Typical "dancing black dots" but living in spheres and with pics!


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#1 Culturific

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Posted 17 July 2011 - 02:21 PM

Been reading through lots of posts about cell contamination, and even found one that had exactly what I was experiencing, but unfortunately there was no resolution. I thought I would post some pics I took to help elucidate the issue. There is a combo of contamination that involves the typical black dots everyone talks about, but also these curious spherical bodies that grow and float around the culture. Sometimes they have large masses attached to them and sometimes they are completely free floating. In both instances, there are dancing black dots on the inside. Dancing bugs are also seen free of these spherical bodies. If I let a culture go without washing of these spherical bodies, the culture will surely be lost and over run with bugs (notin spheres), with high turbidity and pH changes.

Cultures are incubated with Pen/strep mix, but to no avail. I haven't tried other, stronger antibiotics yet. They also seem to be highly contagious from culture to culture, as all of my cell lines have something in them.

Ealier post describing the same here.

I've attached some images. Imagine dancing black dots on the inside.

Some answers I've found might be possible pleuropneumonialike organisms (PPLO)and there are many other possibilities found in this thread.

We've done lots of cleaning: cleaned hoods with 70% ethanol and Conflikt with UV under tray area, decon the Heracell incubators with onboard decon routine (90C in humidified atmosphere for 24hours), stripped, waxed and mopped floors,changed filters on traps, serological pipetors, autoclaved pipet, etc. with no luck so far. There are more methods we're to employ as well, but I'm just mostly curious if anyone knows the biology of the bug in the pics and have had any luck getting rid of it. They seem to be intracellular, but I haven't necessarily seen them inside any live cells, but the spheres seem to come off of what were cells at one time. Dishes can be washed clean of the debris, but it always comes back. Any insight would certainly be appreciated. Thanks for your time!

Attached Thumbnails

  • Sphere.jpg
  • Sphere6.jpg


#2 bob1

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Posted 17 July 2011 - 03:24 PM

The speherical bodies are dead/dying cells.




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