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We have expressed, in vitro (TnT system), mTFP, mCerulean, mCitrine and
mCherry and bound each FP successfully to Protein G agarose beads via an
appropriate antibody. Although each set of beads is easily imaged
individually wed like to use them in combination to investigate whether
either 1) our imaging software (Olympus cellSens Dimension) can effectively
spectrally unmix emissions from mixed bead sets or 2) we can spectrally
discriminate beads using dedicated narrow-pass filter sets.
The beads are 45-165 µm dia. Rather than just randomly mixing the beads are
there slides available with sufficiently small micro-wells so we could see
each FP set separately but in a single field of view?
Also, am I correct in thinking that, as we intend to use 2 or 3 of these FPs
as multiple gene expression reporter tags in individual C. elegans, spectral
unmixing may be the best way to go especially as expression of one or more
target genes may be low?
Locked
