Re: lysosomal live cell markers

Posted by John Naleway on
URL: http://confocal-microscopy-list.275.s1.nabble.com/lysosomal-live-cell-markers-tp7581040p7581047.html

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Dear Jason,

From our experience, the MarkerGeneT LysoLiveT Lysosomal Metabolic Health
Assay Kit http://www.markergene.com/M1910.php , Product M1910 has the best
properties for what seems like your application.

It's been validated versus LAMP1 and LAMP2 staining, is extremely bright and
relatively photostable and exhibits significantly less toxicity than
LysoTracker.  Because it's an enzyme substrate, it will accumulate in the
cells, and some optimization might be needed since it utilizes a kinetic
assay format.

Cheers,

John


----- Original Message -----
From: "Jason Miller" <[hidden email]>
To: <[hidden email]>
Sent: Friday, September 27, 2013 10:26 AM
Subject: lysosomal live cell markers

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> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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>
> Hi all-
>
> I will be doing a series of experiments that involves tracking lysosomes
> by
> live-cell microscopy over 1-24 hours. I took a look at the listserv
> archives and hadn't seen much recently on the topic of live cell lysosomal
> markers. While LysoTracker has been a standard for a while, my
> understanding is that there are potential pitfalls, including purported
> photoswitching from red to green with LysoTracker is excited, potential
> toxicity, lack of specificity to lysosome (rather than just all acidic
> compartments), and low fluorescence levels.
>
> Compared to LysoTracker, how are some of the other non-FP based methods
> for
> live-cell lysosomal tracking? I'd be curious to hear thoughts on Enzo's
> Lyso-ID, MarkerGene's LysoLive lysosomal sulfatase kit, Abcam's
> CytoPainter
> lysosomal kit, AAT's LysoBrite (which they claim is much more photostable
> and less toxic than LysoTracker), Magic Red detection of cathepsin B
> activity, or preloading cells with fluorescently labeled dextrans?
>
> I'm specifically interested in:
> a) specificity of labeling
> b) toxicity
> c) photostability (resistance to bleaching and photoconversion)
> d) how long the label lasts if I were to do a prolonged time-course
> experiment (e.g. 24 hours)?
>
> Thanks so much for your insights in advance.
>
> -Jason Miller
>
> --
>
> Jason Miller, MD, PhD
>
> University of Michigan Kellogg Eye Center
>
> E-mail: *[hidden email] *
>
>
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