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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: *[log in to unmask] *


The first thing which I can record concerning myself is, that I was born.
These are wonderful words. This life, to which neither time nor eternity
can bring diminution - this everlasting living soul, began. My mind loses
itself in these depths.  -- Groucho Marx