Abstract
Fluorescence correlation spectroscopy (FCS) is a powerful technique for measuring low concentrations of fluorescent molecules and their diffusion constants. In the standard case, fluorescence fluctuations are measured in an open detection volume defined by the confocal optics. However, if FCS measurements are carried out in cellular processes that confine the detection volume, the standard FCS model leads to erroneous results. In this paper, we derive a modified FCS model that takes into account the confinement of the detection volume. Using this model, we have carried out the first FCS measurements in dendrites of cultured neurons. We further derive, for the case of confined diffusion, the limits within which the standard two- and three-dimensional diffusion models give reliable results.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3294-3306 |
| Number of pages | 13 |
| Journal | Biophysical Journal |
| Volume | 79 |
| Issue number | 6 |
| State | Published - Dec 2000 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biophysics
Cite this
Fluorescence correlation spectroscopy in small cytosolic compartments depends critically on the diffusion model used. / Gennerich, Arne; Schild, Detlev.
In: Biophysical Journal, Vol. 79, No. 6, 12.2000, p. 3294-3306.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Fluorescence correlation spectroscopy in small cytosolic compartments depends critically on the diffusion model used
AU - Gennerich, Arne
AU - Schild, Detlev
PY - 2000/12
Y1 - 2000/12
N2 - Fluorescence correlation spectroscopy (FCS) is a powerful technique for measuring low concentrations of fluorescent molecules and their diffusion constants. In the standard case, fluorescence fluctuations are measured in an open detection volume defined by the confocal optics. However, if FCS measurements are carried out in cellular processes that confine the detection volume, the standard FCS model leads to erroneous results. In this paper, we derive a modified FCS model that takes into account the confinement of the detection volume. Using this model, we have carried out the first FCS measurements in dendrites of cultured neurons. We further derive, for the case of confined diffusion, the limits within which the standard two- and three-dimensional diffusion models give reliable results.
AB - Fluorescence correlation spectroscopy (FCS) is a powerful technique for measuring low concentrations of fluorescent molecules and their diffusion constants. In the standard case, fluorescence fluctuations are measured in an open detection volume defined by the confocal optics. However, if FCS measurements are carried out in cellular processes that confine the detection volume, the standard FCS model leads to erroneous results. In this paper, we derive a modified FCS model that takes into account the confinement of the detection volume. Using this model, we have carried out the first FCS measurements in dendrites of cultured neurons. We further derive, for the case of confined diffusion, the limits within which the standard two- and three-dimensional diffusion models give reliable results.
UR - http://www.scopus.com/inward/record.url?scp=0033635801&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033635801&partnerID=8YFLogxK
M3 - Article
VL - 79
SP - 3294
EP - 3306
JO - Biophysical Journal
JF - Biophysical Journal
SN - 0006-3495
IS - 6
ER -