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Jeff
We have used Dylight 633 without any noticeable bleaching. We did find that it is pH sensitive though - fluorescence is destroyed at pH9 so we use glycerol in buffer at pH7 for mounting. Our acquisition is over about 1h with 3 sequential excitations for the triple labelling so quite intense irradiation.
Dr Lloyd Donaldson
Project Leader - Microscopy & Wood Identification
Senior Scientist - Plant Cell Walls & Biomaterials
Scion - Forests, Products, Innovation
49 Sala Street, Rotorua 3010
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Ph 07 343 5581
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-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Jeff Spector
Sent: Wednesday, 26 March 2014 10:09 a.m.
To:
[hidden email]
Subject: dyLight 488 for single molecule imaging
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Greetings,
This may be a bit off topic as it is more of a single molecule question, but this list seems to be full of people who are experts in many different imaging modalities, not just confocal so here goes...
A labmate of mine is attempting to use DyLight 488 for some single molecule measurements. We are running into the issue of the molecules bleaching in about the same time as a single GFP molecule bleaches ( we tacked them both down and measured the bleaching times). I was under the impression that the DyLight dyes were supposed to be much brighter and photo-stable than GFP ( at least I thought they were "related" to the Alexa dyes which are). I was just wondering if anyone out there has had experience using dylight 488, especially for single molecule measurements, and what their experiences were..
thanks..
-Jeff
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