Dynamic light scattering => baseline

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The discussion revolves around the challenges faced in using dynamic light scattering (DLS) for quality control of proteoliposomes containing the fluorescent probe pyranine. The user reports that the baseline consistently exceeds 2 when pyranine is present, leading to aberrant autocorrelation curves and unreliable data on liposome characteristics. A suggestion is made to titrate the pyranine to determine if the issue is due to background fluorescence. The consistent baseline level, regardless of pyranine concentration, raises questions about whether the problem lies with the probe itself or another factor. The conversation highlights the need for further investigation into the effects of fluorescent probes on DLS measurements.
Martin Picard
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Hi !
Sorry for a naive question from a biologist. I am making proteoliposomes within which a fluorescent probe, pyranine (lex = 455 nm, lem = 509 nm), has been trapped. I try to quality control my preparations using DLS (Dynapro, Proteinsolutions) and my concern is that whatever the measurement, if pyranin is present, the baseline is systematically above 2 ... Hence the autocorelation curve is aberrant and I can not obtain any reasonable information on my liposomes.I obtain decent results as soon as pyranin is absent. The laser wavelength is 8336 A ...
Does anyone have an idea what is going on ?

Thanks

Martin
 
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Can you titrate the pyranine? I'm just wondering if the problem is background fluorescence or not- if the baseline is always the same level regardless of the amount of pyranine present, it would argue that the problem is not pyranine but something else.
 

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