[Histonet] Re: how to isolate scabs

[Jerry (Gerald) Sedgewick]

   Hello all,
   I  have  received embolic material in test tubes in 10% formalin.  The
   tubes  had  been  sitting around for about 2 years.  To quantitate the
   number  of embolic particles, after removing particles at less than 40
   microns,  the  solution was poured into a petri dish and then scanned.
   However,  in  about 1/3rd of the test tubes, a hard disk of what looks
   like  red  blood  cells formed on the bottom when the RBCs should have
   been  suspended  in solution.  These hard discs (scabs) were broken up
   and also included in the petri dish scans.
   The  problem  is  that the scabs should not be measured with the other
   particles because these weren't measured in a previous study: the RBCs
   in a previous study did not coagulate.
   So now I have to figure out which of the broken up scabs are scabs and
   which  could be other embolic material.  In the past, I irradiated the
   RBCs  with  green  light  and  collected through a red filter and this
   revealed  RBC laden particles.  However, these aren't autofluorescing,
   possibly  because  they  have been in solution so long (or because the
   scabs are a mix of embolic materials).
   Here's  the  million dollar question: has anyone had to identify scabs
   on  a  macro  level,  and  how  was that done?  I know all the embolic
   material  can  be  spun down and sectioned, but the idea is to get the
   quantitative  information  about  the  nature  of the particles BEFORE
   spinning down.
   Thanks!
   Jerry

-- 
Jerry (Gerald) Sedgewick
Author: "Scientific Imaging with Photoshop: Methods, Measurement and Output." 

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References

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   4. http://www.jerrysedgewick.com/