Interesting bacteria structures or problem with over heat fixing?

[Jshirls]

Hi,

I prepared a whole bunch of slides as follows
1. From 3d turbid liquid broth pure culture (soil bacteria) transferred loopful of broth culture onto a slide
2. Let air dry
3. Heat fixed by passing through bunsen burner 5-6 times briefly, specimen facing upwards

Most people seem to pass through very briefly in videos I watched, I went a little longer because I was scared of not actually killing the bacteria, as it seemed overly short in the videos. I still only spent at most a second or two in the flame, and only 5-6 passes. So not 'much' longer. No obvious burning or transformation of what was on top.

My specimens are supposed to be soil bacteria, although they are unknown so I am looking through these pure cultures in a Zeiss el-einsatz microscope at 40x, 100x, 400x and 1000x oil immersion.

I am using Kohler illumination and only oil immersion at 1000x. I did not use a coverslip or any staining. Images taken with cellphone through eyepiece.

I looked at 3 unique samples in the microscope and saw branching shapes and nice patterns. Now, I have not ever looked at these specimens under the microscope so I was not sure what I was 'expecting' to see, but definetly was not expecting to see this, let alone somewhat similar for all 3 specimens. They did come from the same plant and same plant organ, so I guess they could be similar. But they are supposed to bacteria so, I did not expect to see this structures.

Can anyone confirm if these structures are in fact the bacteria with odd morphology that I"m looking at, or if this may be some error or possibly crystallization of the broth medium caused by too long of a heat fix? Maybe someone here has seen this before or has experienced a problem with heat fixing?

Here are the pictures
1. Sample 1 100x https://www.flickr.com/photos/196129368 ... ed-public/
2. Sample 1 400x https://www.flickr.com/photos/196129368 ... ed-public/
3. Sample 2 100x https://www.flickr.com/photos/196129368 ... ed-public/
4. Sample 2 400x https://www.flickr.com/photos/196129368 ... ed-public/
5. Sample 3 100x https://www.flickr.com/photos/196129368 ... ed-public/



Thanks in advance,

Jshirls

[Hobbyst46]

To me they look like crystals of a mineral or organic salt or acid. Not microorganisms.

[Tom Jones]

Years ago, my first microbiology professor said of heat fixing, to pass it through the flame 2-3 times, smear up, then lay the slide on the back of your (then ungloved) hand. If you heard the sound of sizzling flesh, it was too hot!

[Leitzcycler]

Some stuff from your culture medium I think. It is my experience that samples taken directly from culture medium seldom give acceptable results.

1. From culture plates: pick a part of a colony and suspend into a drop of water on slide.
2. Liquid cultures: centrifuge a sample of the culture and discard the supernatant. Resuspend the pellet into water and apply a drop onto slide (or take some of the pellet and proceed as above).

I don't regularly use flaming. I just let the slide to dry onto a heat plate, the temperature about +45. Then staining and washing.