[Cytometry] Merocyanine fluorescence

Anna Petrunkina ap542 at cam.ac.uk
Wed May 6 07:37:34 EDT 2009


Hi Kathy,

merocyanine is a fluorescent lipophilic probe which binds to the plasma 
membrane depending on the degree of the lipid architecture disorder. 
Changes in merocyanine binding (i.e. evolving subpopulations with increased 
intensity) indicates a changes in the  assymetry of the membrane 
phospholipid distribution. In sperm it is thought to be a sign of the 
physiologically induced destabilization, which in vitro will be induced by 
the bicarbonate/CO2. Also, it will be most important to gas your samples 
with CO2 and have adequate controls.

There has been a lot of research done re mero binding in mammallian sperm 
(have not heard much about avian sperm, though). Here are some references 
which describe protocols and ideas quite nicely:

Mol Reprod Dev. 1996 Nov;45(3):378-91.; Mol Reprod Dev. 2000 
Feb;55(2):220-8.; Development. 2000 Jun;127(11):2407-20; Biol Reprod. 2002 
Jul;67(1):340-50.

When using cryopreservation, there may be some severe problems if 
cryodiluent contains egg yolk (as it will almost certainly), so one will 
need to work out a relevant approach of removing diluent.

Hope that helps,
Anna

Dr. Anna Petrunkina
Head of Flow Cytometry
Cambridge Institute for Medical Research
University of Cambridge
Wellcome Trust/MRC Building
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2XY

--On 05 May 2009 11:49 +0800 Kathy Heel <kathy.heel at uwa.edu.au> wrote:

> Hi All,
>
>
>
> I have a researcher interested in analyzing the membrane fluiditiy of
> emu sperm after cryopreservation using merocyanine (membrane fluidity)
> and Yo-Pro-1 (viability). I have a basic understanding of some human
> sperm assays but have not encountered merocyanine before so I'm not
> really sure I understand what the merocyanine is staining. Has anyone
> used merocyanine previously and if so can you give me any tips please?
> Also I don't have any experience with avian samples, so any and all
> suggestions from the avian/sperm flow gurus will be much appreciated.
>
>
>
> Thanks on advance
>
> Kathy
>
>
>
> Kathy Heel PhD
>
> Assistant Professor
>
> Centre for Microscopy, Characterisation and Analysis (M510)
>
> The University of Western Australia
>
> 35 Stirling Highway
>
> Crawley WA 6009
>
> Australia
>
>
>
> Ph 61 8 9346 4525
>
> Fax 61 8 9346 3469
>
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>
> www.cmca.uwa.edu.au
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