http://confocal-microscopy-list.275.s1.nabble.com/Deconvolving-Spinning-Disk-Images-tp6508446p6512630.html
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> Hello Dear Listers,
>
> We have run into a conundrum regarding live, semi-long term and fixable
> labelling of two population of mouse ES cells for aggregation/sorting out
> studies.
> We would like to fix label cells (two cell strains), let them aggregate,
> fix the aggregates after 3-4 days and be able to make them out with either
> am ancient Bio-Rad MRC 600 Krypton/Argon (488, 568 and 647 nm) or
> a prehistorical Zeiss Photomicroscope (DAPI/Hoechst @ ~350 nm, FITC @ ~390
> nm and TRITC (~440 nm) ot Texas Red (~620 nm).
>
> Invitrogen/Molecular Probes (
http://www.invitrogen.com/**> site/us/en/home/References/**Molecular-Probes-The-Handbook/**
> Fluorescent-Tracers-of-Cell-**Morphology-and-Fluid-Flow/**
> Membrane-Permeant-Reactive-**Tracers.html#head2<
http://www.invitrogen.com/site/us/en/home/References/Molecular-Probes-The-Handbook/Fluorescent-Tracers-of-Cell-Morphology-and-Fluid-Flow/Membrane-Permeant-Reactive-Tracers.html#head2>)
> offer quite few cell tracking probes but figuring out what does what is
> difficult (for me, at least).
> They say that CellTracker Green CMFDA (5-chloromethylfluorescein diacetate)
> labels cells that are both fluorescent and viable for at least 24 hours
> after loading.
> We need three-four days, though.
> Orange-fluorescent CellTracker Orange CMRA - ditto.
>
> CellTrace Oregon Green 488 Carboxylic Acid Diacetate Succinimidyl Ester is
> highly recommended but nothing is said of its fixability.
> It is said that CellTrace Far Red DDAO-SE is a fixable, far-red–fluorescent
> tracer for long-term cell labelling but from the literature given I am not
> sure of its longevity.
>
> Sigma (
http://www.sigmaaldrich.com/**img/assets/8200/PKH_LinkerKit_**> web_version.pdf<
http://www.sigmaaldrich.com/img/assets/8200/PKH_LinkerKit_web_version.pdf>)
> sells PKH26 and PKH67 that (Sigma says) are stable for up to 100 days in
> vivo (PKH26), but I do not see them heavily used in cell tracking other than
> FACS.
>
> /Ergo/,
> Any practical suggestion shall be greatly appreciated!
>
> Greetings,
>
> Michal
>
> PS
> Obviously (and to my chagrin) I have no financial association with neither
> company.
> M.O.
>
Joel B. Sheffield, Ph.D