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Hi,

This question just fit in perfectly on what I am trying to find out about
colocalization.

When and why do I need do deconvolve pictures collected with a confocal in
order to be sure about my colocalization (or not colocalization) results?

To be specific: I am working on pre and post-synaptic proteins.

Thanks

Valeria



> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Colocalization based upon "yellow" could be accurate, if and only if,
> the intensities are comparable and pixel (voxel) quantities in the
> suspected colocalized volumes are in roughly equal.  .  Otherwise,
> the yellow is masked by the predominate channel.  Something small,
> like lysosomes, would need to be sampled properly. Colocalization
> could be masked by blur unless deconvolved, even if images are
> collected with a confocal.
> On Feb 7, 2007, at 1:05 PM, Marc Thibault wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Hi all,
>>
>> It seems that in many papers from biologists or chemists, and i'm
>> talking
>> high impact factors journals,  colocalisation of two elements is is
>> often
>> assumed  by simple color superposition (ex: red and green fluoresce
>> yellow
>> when colocalising), while microscopists (many physisists I suppose)
>> seem to
>> need a more complex software-based confirmation.
>> Is it ok, when using high end equipment and corrected objectives
>> (apochromat
>> with high NA for ex.), to assume colocalisation by color
>> superposition,
>> especially when fluorophore are confined to small volume entities,
>> like
>> lysosomes ?
>>
>> Thanks
>>
>> Marc
>