Elaine Coderre wrote:
>
> We have a multi users Bio-Rad MRC1024 Ar/Kr confocal laser facility. My
> problem is that we are trying to set-up the inverted microscope for
> different applications. Some investigators will be looking at live tissue.
> Their needs requires a phase objective.
> Others are strictly looking at fluorochrome (FITC, Texas Red, rhodamine
> dextrans etc...). The 60x oil that we now have does not have phase
> capabilities. My question is how much does a 60x oil with phase  effect
> the resolution of the image for the single fluorochrome users?
> Thanks
> Elaine Coderre
> Loeb research institute
> Ottawa, Ontario
> [log in to unmask] Elaine,

Unless the phase version of the objective has a lower NA than the
conventional 60x, it should not have a big impact on your imaging.  The
phase ring only does its trick when matched with a phase annulus in the
condenser (transmitted light).  Fluorescence works in epi (reflected
light).  You still have essentially the full NA of the objective for the
fluorescence part of the light path.  The only place you may get into
trouble is some reflection of fluorescence off the edge of the cut in the
phase plate.  My best suggestion .... try it with your particular sample.

Best of luck.

Barbara Foster
MME