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Why my bacteria cells look weird in DAPI fluorescence?

immunofluorescence bacteria

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#1 Meg P. Anula

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Posted 28 March 2016 - 05:01 AM

Hi all,

 

Are bacteria cells supposed to look smaller and not in their own shape when in fluorescence?

 

I did DAPI fluorescence stain to view my bacteria, I did not see any single bacillus like in the slide. I saw what supposed to be cell clump, but in it the cell that fluoresces looks so weird, it was like little spots. Overall, I just could not see bacillus shaped creatures. All I see was cell clumps (attached in pic here).

 

I did gram stain on my bacteria cells and confirm it is indeed bacillus. 

 

I use x40 on both. (maximum on the microscope)

 

Attached are the pictures.

 

Pls help.

Attached Thumbnails

  • gram stain bacillus (jpeg).jpg
  • site 5 dapi x40 50ms (jpeg).jpg


#2 Leishman001

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Posted 29 March 2016 - 11:39 AM

When you stain with DAPI, only the nuclei will fluoresce, as DAPI associates with DNA. (https://en.wikipedia.org/wiki/DAPI). The cell body or outline will not be seen or will appear very faint compared to the much smaller nuclei which will fluoresce brightly.



#3 mdfenko

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Posted 30 March 2016 - 03:33 AM

keep in mind that meg p. anula is staining bacteria. no nucleus. however, the dna is stained, not the membrane or wall.


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