Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalNicole - Are you looking at eukaryotic cells infected with amoebae
AND you want to see bacteria in this sample as well? If so, my
experience is that, at least for non-living eukaryotic cells, you
should be able to see nuclei using the BacLight stain. The PI should
show the nuclei and, if this isn't working, you could try acridine
orange - we have even seen Syto 9 staining of nuclei in shed cheek
epithelial cells (with pretty bacteria stuck all over them).
However, anything you add besides the BacLight (including, for
example, acridine orange) will stain your bacteria as well.
Basically, I am surprised that you see no nuclear staining in a
sample exposed to BacLight.
If the "viability" part is of less importance than just seeing the
bacteria, then acridine orange should work just fine for nuclei as
well as bugs.
>Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
>I am tryring to view an eukaryotic cell infected with amoeba and I
>am using BacLight for the bacteria but it is also staining the
>cytoplasm of my amoeba green making it difficult to determine the
>location of the live bacteria. Can anyone recommend a dye that
>would be better or a dye that would stain the amoeba a different
>color other than green? The lasers I have to work with range from
>457 to 639 so DAPI and Hoescht are out.
>
>Nicole Young
--
Robert J. Palmer Jr., Ph.D.
Natl Inst Dental Craniofacial Res - Natl Insts Health
Oral Infection and Immunity Branch
Bldg 30, Room 310
30 Convent Drive
Bethesda MD 20892
ph 301-594-0025
fax 301-402-0396
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