Abstract
A fluorescence quenching competitive immunoassay in micro droplets was applied to the sensitive detection of the pyrethroid insecticide, esfenvalerate. Laser induced fluorescence from rhodamine dye was used as a marker. The competitive immunoreaction was performed in micro droplets generated by a vibrating orifice aerosol generator system with a 10-μm diameter orifice. Fluorescence that was emitted from the droplets was detected by a 1/8 m imaging spectrograph with a 512×512 thermoelectrically cooled, charged-coupled device camera. The conjugate of esfenvalerate with rhodamine exhibited similar fluorescence to that of pure rhodamine 6G. When anti-esfenvalerate antibodies were added to the droplets, the fluorescence decreased. The reduction in emission was due to a strong quenching effect that arises from the interaction between the protein and rhodamine molecules following the antigen-antibody reaction. When a sample of esfenvalerate was added to the droplets, the release of the conjugated rhodamine from the antigen-antibody complex allowed the fluorescence signal to recover. An assay in a picoliter droplet sample was shown to enable detection down to approximately 0.1 nM. A very small mass of analyte could be detected with this method. A sample of river water was used to gauge the impact of matrix effects and was shown to give rise to negligible interference with the immunoassay.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1055-1063 |
| Number of pages | 9 |
| Journal | Biosensors and Bioelectronics |
| Volume | 18 |
| Issue number | 8 |
| DOIs | |
| State | Published - Aug 1 2003 |
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Keywords
- Esfenvalerate
- Fluorescence
- Immunoassay
- Micro droplet
- Quenching
ASJC Scopus subject areas
- Biotechnology
- Analytical Chemistry
- Electrochemistry
Cite this
Fluorescence quenching competitive immunoassay in micro droplets. / Feng, Jun; Shan, Guomin; Hammock, Bruce D.; Kennedy, Ian M.
In: Biosensors and Bioelectronics, Vol. 18, No. 8, 01.08.2003, p. 1055-1063.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Fluorescence quenching competitive immunoassay in micro droplets
AU - Feng, Jun
AU - Shan, Guomin
AU - Hammock, Bruce D.
AU - Kennedy, Ian M.
PY - 2003/8/1
Y1 - 2003/8/1
N2 - A fluorescence quenching competitive immunoassay in micro droplets was applied to the sensitive detection of the pyrethroid insecticide, esfenvalerate. Laser induced fluorescence from rhodamine dye was used as a marker. The competitive immunoreaction was performed in micro droplets generated by a vibrating orifice aerosol generator system with a 10-μm diameter orifice. Fluorescence that was emitted from the droplets was detected by a 1/8 m imaging spectrograph with a 512×512 thermoelectrically cooled, charged-coupled device camera. The conjugate of esfenvalerate with rhodamine exhibited similar fluorescence to that of pure rhodamine 6G. When anti-esfenvalerate antibodies were added to the droplets, the fluorescence decreased. The reduction in emission was due to a strong quenching effect that arises from the interaction between the protein and rhodamine molecules following the antigen-antibody reaction. When a sample of esfenvalerate was added to the droplets, the release of the conjugated rhodamine from the antigen-antibody complex allowed the fluorescence signal to recover. An assay in a picoliter droplet sample was shown to enable detection down to approximately 0.1 nM. A very small mass of analyte could be detected with this method. A sample of river water was used to gauge the impact of matrix effects and was shown to give rise to negligible interference with the immunoassay.
AB - A fluorescence quenching competitive immunoassay in micro droplets was applied to the sensitive detection of the pyrethroid insecticide, esfenvalerate. Laser induced fluorescence from rhodamine dye was used as a marker. The competitive immunoreaction was performed in micro droplets generated by a vibrating orifice aerosol generator system with a 10-μm diameter orifice. Fluorescence that was emitted from the droplets was detected by a 1/8 m imaging spectrograph with a 512×512 thermoelectrically cooled, charged-coupled device camera. The conjugate of esfenvalerate with rhodamine exhibited similar fluorescence to that of pure rhodamine 6G. When anti-esfenvalerate antibodies were added to the droplets, the fluorescence decreased. The reduction in emission was due to a strong quenching effect that arises from the interaction between the protein and rhodamine molecules following the antigen-antibody reaction. When a sample of esfenvalerate was added to the droplets, the release of the conjugated rhodamine from the antigen-antibody complex allowed the fluorescence signal to recover. An assay in a picoliter droplet sample was shown to enable detection down to approximately 0.1 nM. A very small mass of analyte could be detected with this method. A sample of river water was used to gauge the impact of matrix effects and was shown to give rise to negligible interference with the immunoassay.
KW - Esfenvalerate
KW - Fluorescence
KW - Immunoassay
KW - Micro droplet
KW - Quenching
UR - http://www.scopus.com/inward/record.url?scp=0037681599&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037681599&partnerID=8YFLogxK
U2 - 10.1016/S0956-5663(02)00218-X
DO - 10.1016/S0956-5663(02)00218-X
M3 - Article
C2 - 12782469
AN - SCOPUS:0037681599
VL - 18
SP - 1055
EP - 1063
JO - Biosensors
JF - Biosensors
SN - 0956-5663
IS - 8
ER -