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>>Hi all,
>>
>>
>>We are looking at performing a quantitative blood flow analysis by
counting fluorescent microspheres (15 microns size, made of polystyrene and
coated with flourescent dye) in the nerve of a rat after injection into the
abdominal aorta.
>>
>>We would appreciate some advice on the counting method:
>>
>>Here is what we have tried so far.
>>
>>our nerve section is 3cm long.  We tried using a fluorescent microscope to
count the spheres but that was quite difficult due to the 3D distribution of
the spheres, and the limited field of view especially at the requires 100x
mag resulting in overlap in counting of the spheres.
>>
>>Other options include:
>>
>>digesting  the tissue to extract the dye from the spheres and then using a
fluorometer but we believe the low concentration of the spheres in our
tissue amy be the limiting factor.
>>
>>filtering the solution containing the digested tissue, and analysing the
filter under a microscope.
>>
>>If you have any other suggestions, it would be greatly appreciated.
>>
>>Regards,
>>
>>Andras
>>
>>
>>**********************************************
>Tim Cushway          [log in to unmask]
>>Research Fellow
>>Microsearch FOundation of Australia
>>11 Sirius Rd
>>Lane Cove, NSW Australia 2066
>>ph: (+612) 9418 6656  fax: (+612) 9418 9433
>>
>>*********************************************
>>
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