I am looking for an inexpensive and easy to use fluorescent stain for observing Rhizobium cell wall. I am treating this bacterium with a chemical stressor and my metabolomics data shows that the cell wall biosynthesis is compromised. I was wondering if anyone has any suggestions on if/how I can directly visualize this using fluorescent microscopy.
Hi Supriya,
This will probably sound subjective, but we have recently reported molecules that incorporate into sites of active peptidoglycan growth in diverse bacteria.
(https://www.researchgate.net/publication/232231865_In_Situ_Probing_of_Newly_Synthesized_Peptidoglycan_in_Live_Bacteria_with_Fluorescent_D-Amino_Acids)
The probes only stain the active cells, so they distinguish from LIVE/DEAD staining in that regard, which is advantageous for many applications. Not sure what you name as inexpensive but we would gladly send you 2 mg HADA for $250 (enough for ~20 experiments). We are charging this amount in order to cover the cost of synthesis. If you have the means we would gladly share the protocols to synthesize the probes yourself!
By the way, in the paper we worked with Agrobacterium as well, which grows from one tip!
Article In Situ Probing of Newly Synthesized Peptidoglycan in Live B...
You could try some LIVE/DEAD staining - it distinguishes between intact and compromised cell membranes but might correlate with your cell-wall-related metabolomics data too.
Hi Supriya,
This will probably sound subjective, but we have recently reported molecules that incorporate into sites of active peptidoglycan growth in diverse bacteria.
(https://www.researchgate.net/publication/232231865_In_Situ_Probing_of_Newly_Synthesized_Peptidoglycan_in_Live_Bacteria_with_Fluorescent_D-Amino_Acids)
The probes only stain the active cells, so they distinguish from LIVE/DEAD staining in that regard, which is advantageous for many applications. Not sure what you name as inexpensive but we would gladly send you 2 mg HADA for $250 (enough for ~20 experiments). We are charging this amount in order to cover the cost of synthesis. If you have the means we would gladly share the protocols to synthesize the probes yourself!
By the way, in the paper we worked with Agrobacterium as well, which grows from one tip!
Article In Situ Probing of Newly Synthesized Peptidoglycan in Live B...
Hey Erkin!
Your paper sounds really interesting. Would it be possible to probe the bacteria when they are already in one suspension with eukaryotic cells?
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