Hi List,
I think this has been discussed in the past but i have not been able to
find a definitive answer to the problem.
Basically if you have a sample stained with DAPI, after viewing it with
a DAPI filter, the signal can be then detected using a GFP/FITC filter.
I have tried this on samples with nothing but DAPI on them and it still
happens. Suggestions in the past have been to lower the concentration of
DAPI used and to scan/capture the other channels first and DAPI last.
But does anyone out there have any idea why this happens in the first
place? If you look at the spectra of DAPI it is a very good green
emitter (but needs to be excited in the UV-Blue range). It should not be
able to be excited by standard GFP type excitation at 480-490nm. So the
conversion that is happening is shifting the excitation spectra of DAPI
towards the red somehow.
Any ideas?
Cheers
Cam
Cameron J. Nowell
Microscopy Manager
Centre for Advanced Microscopy
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Victoria, 3050
AUSTRALIA
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]]
On Behalf Of Jerry Sedgewick
Sent: Tuesday, 5 May 2009 1:35 AM
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Subject: Re: Posting for Basic Confocal Workshop at U of South Carolina
There are still some slots available in this year's Basic Confocal
Workshop hosted by the University of South Carolina. This year's
workshop will be from June 15-19, 2009 and will include a series of
lectures on the theory and applications of confocal microscopy, specimen
preparation, processing confocal images in Photoshop, and 3D
reconstructions using AMIRA. Students will be able to process triple
labeled samples (cell cultures and sections) on site or bring their own
samples to the workshop.
Faculty will include Drs. Jay Jerome (Vanderbilt), Ralph Albrecht (Univ
Wisconsin-Madison), John Mackenzie (North Carolina State Univ), Tom
TrusK (Medical Univ South Carolina) and myself.
Instruments and applications experts from Leica, Nikon, Olympus, Perkin
Elmer, Photometrics, and Zeiss will be available for hands on training
and imaging of samples.
For those contemplating instrumentation proposals as part of the
stimulus or other funding opportunities this is an excellent opportunity
to see several systems side by side and to collect preliminary data on
their instrument of choice.
For further information and registration go to:
http://dba.med.sc.edu/irf/price/irf/irf.htm or contact Anna McNeal
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Bob
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