Isolation of chloroplasts and Cellquest question

From: Carol Oxford (cloxford@UCDAVIS.EDU)
Date: Thu Jan 20 2000 - 18:45:15 EST


I've been approached by an investigator who wants to separate mesophyl
chloroplasts from bundle sheath chloroplasts in maize, and this is way out
of my area of expertise.  They gave me a reference with Bob Ashcroft's name
on it, so I was hoping to get some advice from the expert!  The papers were
fairly old and detailed several methods including the use of differences in
scatter and in fluorescence emission in the 685-710 range to isolate them.
They also used ratios of fluorescence above and below 710, which seems to
relate to the emission difference between chlorophylls A and B.  Is this
necessary (as we have no filter currently to do this) or is there another
way to do this?  Is anyone out there doing something similar?

My second question came to mind with all of the recently discussed software
issues.  I have a four color XL in the lab, and one of the investigators
using it wants to analyze the files with Cellquest.  I saved the data in
list mode, then transferred to a PC disk.  My G3 could see them, but
Cellquest could not.  Any hints?

Thanks,

Carol






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Carol Oxford                             _|    _|   _|_|_|_|   _|_|_|
Manager, UC Davis Optical Biology Lab   _|    _|   _|         _|    _|
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cloxford@ucdavis.edu               University of California, Davis
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