Dynamic light scattering => baseline

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SUMMARY

The discussion centers on the challenges faced by a biologist using Dynamic Light Scattering (DLS) with a fluorescent probe, pyranine, in proteoliposome preparations. The user reports that the baseline consistently exceeds 2 when pyranine is present, leading to aberrant autocorrelation curves and unreliable data. When pyranine is absent, the DLS results are satisfactory. The conversation suggests investigating the potential for background fluorescence rather than solely attributing the issue to pyranine.

PREREQUISITES
  • Understanding of Dynamic Light Scattering (DLS) principles
  • Familiarity with fluorescent probes, specifically pyranine
  • Knowledge of autocorrelation curve analysis
  • Experience with quality control in biophysical measurements
NEXT STEPS
  • Investigate background fluorescence effects in DLS measurements
  • Learn about titration techniques for fluorescent probes
  • Explore DLS data interpretation methods for liposome characterization
  • Research troubleshooting strategies for DLS baseline issues
USEFUL FOR

This discussion is beneficial for biologists, biophysicists, and researchers involved in liposome characterization and those utilizing Dynamic Light Scattering for quality control in biophysical experiments.

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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