research.gif (2985 bytes) Dr. Gary Coté, Professor of Biology

Isolation of Living Crystal Idioblasts

We have worked out methods to get live biforine idioblasts and mesophyll cells out of leaves.  Other crystal idioblast cells are also isolated, but are apparently not alive.  The method involves digesting the connections between cell walls with enzymes called pectinases and then isolating the cells by density gradient centrifugation.  At right, Jeremy Hensley and Raymond Hein are beginning the process -- notice the leaf pieces in the flasks.  Below center is a picture of the gradient centrifugation.  At right below is an isolated idioblast.  At left below are isolated mesophyll cells.
 
   
Justin Salibrici and Clay Backus made the first attempts to isolate these cells.  Chris Gowdy got the procedure to work, and Mike deMilt greatly improved it.  Brian Fritz, and, of course Jeremy and Raymond, shown above, refined it further.  Jeremy later developed a method that produces far more intact, living biforine idioblasts, at less cost and in much less time.  However, all mesophyll die in this procedure.

Our ability to isolate living mesophyll and living biforines is the foundation for many of our other projects, so this has been one of the most important projects we have undertaken.



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