>
> Hello,
>
> I have been doing some E.Coli analysis and sorting and was
> wondering if anyone had a favorite sheath fluid to use for these
> bugs. I am looking to make them as happy as possible during sort
> cloning. Thanks for any tips.
Many of the commercial preps are unsuitable for bacteria because of
the high sodium content - they are designed to keep animal (blood)
cells happy. I haven't done sorting with coli but Micrococcus luteus
seem ok (i.e. grow afterwards) when sorted in a sheath fluid
containing 150mM KCl and 10mM Hepes (pH 6.8 with KOH). This was with
cells that weren't exactly happy to begin with :
Kaprelyants AS, Mukamolova GV, Davey HM, and Kell DB: Quantitative
analysis of the physiological heterogeneity within starved cultures of
Micrococcus luteus using flow cytometry and cell sorting. Appl. Env.
Microbiol. 62:1311-1316, 1996.
Haze
----------------------------------------------------------
| Hazel Marie Davey hlr@aber.ac.uk |
|Sefydliad y Gwyddorau Biolegol*Inst. Biological Sciences|
|Prifysgol Cymru * University of Wales|
| ABERYSTWYTH, Ceredigion, CYMRU / WALES SY23 3DA |
| http://pcfcij.dbs.aber.ac.uk/index.htm |