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Fast and Sensitive Interferon-γ Assay Using Supercritical Angle Fluorescence.

Winterflood CM, Ruckstuhl T, Seeger S - Biosensors (Basel) (2013)

Bottom Line: We present an immunoassay for Interferon-γ (IFN-γ) with a limit of detection of 1.9 pM (30 pg/mL) and a linear concentration range spanning three orders of magnitude.The solid-phase sandwich assay is performed on a new measurement system comprising single-use test tubes and a compact fluorescence reader.The polymer tubes contain an optical configuration for the detection of supercritical angle fluorescence, allowing for highly sensitive real-time binding measurements.

View Article: PubMed Central - PubMed

Affiliation: Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland; E-Mails: c.winterflood@pci.uzh.ch (C.M.W.); t.ruckstuhl@pci.uzh.ch (T.R.).

ABSTRACT
We present an immunoassay for Interferon-γ (IFN-γ) with a limit of detection of 1.9 pM (30 pg/mL) and a linear concentration range spanning three orders of magnitude. The developed one-step assay takes only 12 min and can replace the time-consuming and labor-intensive enzyme-linked immunosorbent assay (ELISA). The solid-phase sandwich assay is performed on a new measurement system comprising single-use test tubes and a compact fluorescence reader. The polymer tubes contain an optical configuration for the detection of supercritical angle fluorescence, allowing for highly sensitive real-time binding measurements.

No MeSH data available.


Supercritical angle fluorescence (SAF) emission at the solution/substrate interface. (a) The relative SAF collection efficiency of the parabolic element with increasing surface distance z, expressed as a fraction of the emission wavelength, λ. (b) Polar plots of the angular emission distribution of a fluorophore with distances z = 0 (red, solid) and z = λ/3 (blue, dashed) from the interface. Adapted from [14].
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biosensors-03-00108-f001: Supercritical angle fluorescence (SAF) emission at the solution/substrate interface. (a) The relative SAF collection efficiency of the parabolic element with increasing surface distance z, expressed as a fraction of the emission wavelength, λ. (b) Polar plots of the angular emission distribution of a fluorophore with distances z = 0 (red, solid) and z = λ/3 (blue, dashed) from the interface. Adapted from [14].

Mentions: We present a one-step sandwich immunoassay for the quantification of recombinant mouse IFN-γ (rmIFN-γ) within minutes. The assay has been developed on the supercritical angle fluorescence (SAF) immunodiagnostic system, a recently reported fluorescence-based detection platform comprising single-use test tubes and a compact fluorescence reader [14]. The mass producible polymer tubes contain an innovative optical configuration for the collection of SAF, which is emitted above the critical angle into the solid substrate. As shown in Figure 1, SAF occurs only for fluorescent molecules located near the surface (<200 nm). This property of SAF finds application in high resolution microscopy [15,16] or the monitoring of binding reactions in real-time at surfaces [17,18,19].


Fast and Sensitive Interferon-γ Assay Using Supercritical Angle Fluorescence.

Winterflood CM, Ruckstuhl T, Seeger S - Biosensors (Basel) (2013)

Supercritical angle fluorescence (SAF) emission at the solution/substrate interface. (a) The relative SAF collection efficiency of the parabolic element with increasing surface distance z, expressed as a fraction of the emission wavelength, λ. (b) Polar plots of the angular emission distribution of a fluorophore with distances z = 0 (red, solid) and z = λ/3 (blue, dashed) from the interface. Adapted from [14].
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263590&req=5

biosensors-03-00108-f001: Supercritical angle fluorescence (SAF) emission at the solution/substrate interface. (a) The relative SAF collection efficiency of the parabolic element with increasing surface distance z, expressed as a fraction of the emission wavelength, λ. (b) Polar plots of the angular emission distribution of a fluorophore with distances z = 0 (red, solid) and z = λ/3 (blue, dashed) from the interface. Adapted from [14].
Mentions: We present a one-step sandwich immunoassay for the quantification of recombinant mouse IFN-γ (rmIFN-γ) within minutes. The assay has been developed on the supercritical angle fluorescence (SAF) immunodiagnostic system, a recently reported fluorescence-based detection platform comprising single-use test tubes and a compact fluorescence reader [14]. The mass producible polymer tubes contain an innovative optical configuration for the collection of SAF, which is emitted above the critical angle into the solid substrate. As shown in Figure 1, SAF occurs only for fluorescent molecules located near the surface (<200 nm). This property of SAF finds application in high resolution microscopy [15,16] or the monitoring of binding reactions in real-time at surfaces [17,18,19].

Bottom Line: We present an immunoassay for Interferon-γ (IFN-γ) with a limit of detection of 1.9 pM (30 pg/mL) and a linear concentration range spanning three orders of magnitude.The solid-phase sandwich assay is performed on a new measurement system comprising single-use test tubes and a compact fluorescence reader.The polymer tubes contain an optical configuration for the detection of supercritical angle fluorescence, allowing for highly sensitive real-time binding measurements.

View Article: PubMed Central - PubMed

Affiliation: Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland; E-Mails: c.winterflood@pci.uzh.ch (C.M.W.); t.ruckstuhl@pci.uzh.ch (T.R.).

ABSTRACT
We present an immunoassay for Interferon-γ (IFN-γ) with a limit of detection of 1.9 pM (30 pg/mL) and a linear concentration range spanning three orders of magnitude. The developed one-step assay takes only 12 min and can replace the time-consuming and labor-intensive enzyme-linked immunosorbent assay (ELISA). The solid-phase sandwich assay is performed on a new measurement system comprising single-use test tubes and a compact fluorescence reader. The polymer tubes contain an optical configuration for the detection of supercritical angle fluorescence, allowing for highly sensitive real-time binding measurements.

No MeSH data available.