I have a bacterial culture which was grown in a metal containing media. It was observed that, with the time a black color compound was appeared in the media. I want to confirm that whether this compound is, in the media or inside the bacteria. Do anyone know a way to identify such a compound and to confirm the location it present? (Note: I do not have TEM facility)
Which metal containing media was that?
Do you have the original stock of the bacteria?
If you have original stock then try growing both on nutrient agar and let me know your observation.
Most probably it is the metabolic end product (for example H2S on a media containing Sulphur with H2S indicator).
So the question is which media did you use?
Which bacteria is yours, did you identify?
hi, Godfred A Menezes,
Metal - Cu
Media - LB
Bacteria - Unknown
I'm bit confuse about your statement, "If you have original stock then try growing both on nutrient agar and let me know your observation".
What kind of observation do you need? What do you mean by 'both'?
Well if your aim is to observe the color is present in the bacteria, then its simple.... Make the same media in agar form and then spot inoculate the culture onto to the agar. If the copper is taken inside the cell then the cell will show darker color than the media.
Did you grow on any other media initially (if stocked from there you can call it as original stock) or only on LB containing Cu?
If you have grown only on LB containing Cu, then subculture from there onto LB agar (with no Cu) and also Nutrient agar and let me know if your colonies are still dark/ black?
Aspect of non-inoculated plates? How much Cu? What salt did you added? Inoculation technique: streak or mixed with melted agar? Plates or broth?
For the mineralization, the best would be XRF and then XRD. You can use a SEM with EDS but you get only elemental analysis.
@Godfred,
I appreciate your help Godfred. Bacteria was in the liquid broth culture not in solid media. I need to separate bacteria cells, particles(if present outside of the cell) which in the liquid media. How can I separate them?
@ Neethu,
Thank you Neethu, yes when cu is present bacteria is getting dark color. But i want to separate that compound from the bacteria. How can I do that?
This may be a little bit obvious so you may have already tried it. Have you tried doing serial dilutions of the liquid culture in sterile liquid broth, plating dilutions on your simple media agar plates using a melted soft agar over lay, incubate and finally taking a single isolated colony and streaking it using an innocculating loop onto simple media agar plates using a four streak pattern to allow for isolated colonies to grow after incubation along the final serial dilution streak, ?
Instead of plating the dilutions using a melted soft agar overlay you can plate the dilutions directly onto the plate. For example the ten to the minus 5 dilution use 100 microliters and place directly onto the agar plate. Spread with a sterile glass "hockey stick" evenly around the plate (a plate spreader and turntable are useful in attaining a uniform distribution of colonies).
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