double staining test.

[Ozana]

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Hello,

I have a couple ofquestions about a double staining test.

1)  Can prokaryotic cells (gram negative - E. coli and gram pozitive – S. aureus) be stained with acridine orange/ ethidium bromide acetate solution (pH 4), or acridine orange/ethidium bromide  aqueous  solution. There is one of them, none, or both working?2) Another way to immobilize bacteria without killing, besides the polycarbonate membrane and the coated slide with 0.5% agar ?


Thank you!Ozana Petraru – Masterʼ s student in Microbial and cells biotechnologies

 Faculty of Biology

"Al. I. Cuza" Iasi University


 

 

[mmodel]

Hi Ozana,

I don't know how to answer your question about staining, but immobilization of some bacteria can be done by keeping a coverslip in a culture broth for a few hours. Bacteria form a thin biofilm on the glass and get stuck there.

Mike Model

________________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Ozana Maria Petraru <[hidden email]>
Sent: Saturday, April 2, 2016 3:33 AM
To: [hidden email]
Subject: double staining test.

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hello,

I have a couple ofquestions about a double staining test.

1)  Can prokaryotic cells (gram negative - E. coli and gram pozitive – S. aureus) be stained with acridine orange/ ethidium bromide acetate solution (pH 4), or acridine orange/ethidium bromide  aqueous  solution. There is one of them, none, or both working?2) Another way to immobilize bacteria without killing, besides the polycarbonate membrane and the coated slide with 0.5% agar ?


Thank you!Ozana Petraru – Masterʼ s student in Microbial and cells biotechnologies

 Faculty of Biology

"Al. I. Cuza" Iasi University

[Lloyd Donaldson]

Ozana

You can use polylysine or silane coated microscope slides. We normally allow a drop of suspension to dry on the slide followed by staining/washing with acridine orange. Keep in mind the acridine orange will kill the bacteria anyway so drying is not a disadvantage.


Dr Lloyd Donaldson
Microscopy & Wood Identification
Senior Scientist – Plant Cell Walls & Biomaterials
Scion – Forests, Products, Innovation
49 Sala Street, Rotorua 3010
New Zealand
Ph 07 343 5581
www.scionresearch.com



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ozana Maria Petraru
Sent: Saturday, April 02, 2016 8:33 PM
To: [hidden email]
Subject: double staining test.

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hello,

I have a couple ofquestions about a double staining test.

1)  Can prokaryotic cells (gram negative - E. coli and gram pozitive – S. aureus) be stained with acridine orange/ ethidium bromide acetate solution (pH 4), or acridine orange/ethidium bromide  aqueous  solution. There is one of them, none, or both working?2) Another way to immobilize bacteria without killing, besides the polycarbonate membrane and the coated slide with 0.5% agar ?


Thank you!Ozana Petraru – Masterʼ s student in Microbial and cells biotechnologies

 Faculty of Biology

"Al. I. Cuza" Iasi University





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