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Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Földes-Papp, Z; Liao, SC; You, T; Barbieri, B.
Reducing background contributions in fluorescence fluctuation time-traces for single-molecule measurements in solution.
Curr Pharm Biotechnol. 2009; 10(5):532-542
Doi: 10.2174/138920109788922137
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- Földes-Papp Zeno
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- Abstract:
- We first report on the development of new microscope means that reduce background contributions in fluorescence fluctuation methods: i) excitation shutter, ii) electronic switches, and iii) early and late time-gating. The elements allow for measuring molecules at low analyte concentrations. We first found conditions of early and late time-gating with time-correlated single-photon counting that made the fluorescence signal as bright as possible compared with the fluctuations in the background count rate in a diffraction-limited optical set-up. We measured about a 140-fold increase in the amplitude of autocorrelated fluorescence fluctuations at the lowest analyte concentration of about 15 pM, which gave a signal-to-background advantage of more than two-orders of magnitude. The results of this original article pave the way for single-molecule detection in solution and in live cells without immobilization or hydrodynamic/electrokinetic focusing at longer observation times than are currently available.
- Find related publications in this database (using NLM MeSH Indexing)
- Absorptiometry, Photon -
- Algorithms -
- Fluorescence -
- Lasers -
- Solutions - chemistry
- Spectrometry, Fluorescence -
- Time Factors -
- Water -
- Find related publications in this database (Keywords)
- Single-molecule fluorescence fluctuation spectroscopy and imaging
- solution
- fluorescence correlation
- FCS
- scattered background
- Raman and Rayleigh stray light
- autofluorescence
- pulsed excitation
- modulated excitation
- detector gating
- time-correlated single-photon counting
- TCSPC
- early and late time gating
- fluorescence lifetime