Abstract
Current organic fluorophores used as labeling reagents for biomolecule conjugation have significant limitations in photostability. This compromises their performance in applications that require a photostable fluorescent reporting group. For example, in molecular imaging and single molecule microscopy, photostable fluorescent labels are important for observing and tracking individual molecular events over extended period of time. We report in this paper an extremely photostable and highly fluorescent phthalocyanine dye, IRDye™ 700DX, as a near-infrared fluorescence labeling reagent to conjugate with biomolecules. This novel water-soluble silicon phthalocyanine dye has an isomericly pure chemical structure. The dye is about 45 to 128 times more photostable than current near-IR fluorophores, e.g. Alexa Fluor® 680, Cy™ 5.5, Cy™7 and IRDye™ 800CW dyes; and about 27 times more photostable than tetramethylrhodamine (TMR), one of the most photostable organic dyes. This dye also meets all the other stringent requirements as an ideal fluorophore for biomolecules labeling such as excellent water solubility, no aggregation in high ionic strength buffer, large extinction coefficient and high fluorescent quantum yield. Antibodies conjugated with IRDye™ 700DX at high D/P ratio exist as monomeric species in high ionic buffer and have bright fluorescence. The IRDye™ 700DX conjugated antibodies generate sensitive, highly specific detection with very low background in Western blot and cytoblot assays.
| Original language | English (US) |
|---|---|
| Title of host publication | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| DOIs | |
| State | Published - May 8 2006 |
| Event | Optical Molecular Probes for Biomedical Applications - San Jose, CA, United States Duration: Jan 22 2006 → Jan 24 2006 |
Publication series
| Name | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
|---|---|
| Volume | 6097 |
| ISSN (Print) | 1605-7422 |
Conference
| Conference | Optical Molecular Probes for Biomedical Applications |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 1/22/06 → 1/24/06 |
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Keywords
- Bioconjugate
- Cytoblot assay
- Immunoblotting assay
- Isomericly pure
- Near-infrared fluorescence
- Photobleaching quantum yield
- Photostable dye
- Water-soluble phthalocyanine dye
ASJC Scopus subject areas
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Atomic and Molecular Physics, and Optics
- Radiology Nuclear Medicine and imaging
Cite this
Phthalocyanine dye as an extremely photostable and highly fluorescent near-infrared labeling reagent. / Peng, Xinzhan; Draney, Daniel R.; Volcheck, William M.; Bashford, Gregory R.; Lamb, Donald T.; Grone, Daniel L.; Zhang, Yonghong; Johnson, Craig M.
Progress in Biomedical Optics and Imaging - Proceedings of SPIE. 2006. 60970E (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 6097).Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Phthalocyanine dye as an extremely photostable and highly fluorescent near-infrared labeling reagent
AU - Peng, Xinzhan
AU - Draney, Daniel R.
AU - Volcheck, William M.
AU - Bashford, Gregory R.
AU - Lamb, Donald T.
AU - Grone, Daniel L.
AU - Zhang, Yonghong
AU - Johnson, Craig M.
PY - 2006/5/8
Y1 - 2006/5/8
N2 - Current organic fluorophores used as labeling reagents for biomolecule conjugation have significant limitations in photostability. This compromises their performance in applications that require a photostable fluorescent reporting group. For example, in molecular imaging and single molecule microscopy, photostable fluorescent labels are important for observing and tracking individual molecular events over extended period of time. We report in this paper an extremely photostable and highly fluorescent phthalocyanine dye, IRDye™ 700DX, as a near-infrared fluorescence labeling reagent to conjugate with biomolecules. This novel water-soluble silicon phthalocyanine dye has an isomericly pure chemical structure. The dye is about 45 to 128 times more photostable than current near-IR fluorophores, e.g. Alexa Fluor® 680, Cy™ 5.5, Cy™7 and IRDye™ 800CW dyes; and about 27 times more photostable than tetramethylrhodamine (TMR), one of the most photostable organic dyes. This dye also meets all the other stringent requirements as an ideal fluorophore for biomolecules labeling such as excellent water solubility, no aggregation in high ionic strength buffer, large extinction coefficient and high fluorescent quantum yield. Antibodies conjugated with IRDye™ 700DX at high D/P ratio exist as monomeric species in high ionic buffer and have bright fluorescence. The IRDye™ 700DX conjugated antibodies generate sensitive, highly specific detection with very low background in Western blot and cytoblot assays.
AB - Current organic fluorophores used as labeling reagents for biomolecule conjugation have significant limitations in photostability. This compromises their performance in applications that require a photostable fluorescent reporting group. For example, in molecular imaging and single molecule microscopy, photostable fluorescent labels are important for observing and tracking individual molecular events over extended period of time. We report in this paper an extremely photostable and highly fluorescent phthalocyanine dye, IRDye™ 700DX, as a near-infrared fluorescence labeling reagent to conjugate with biomolecules. This novel water-soluble silicon phthalocyanine dye has an isomericly pure chemical structure. The dye is about 45 to 128 times more photostable than current near-IR fluorophores, e.g. Alexa Fluor® 680, Cy™ 5.5, Cy™7 and IRDye™ 800CW dyes; and about 27 times more photostable than tetramethylrhodamine (TMR), one of the most photostable organic dyes. This dye also meets all the other stringent requirements as an ideal fluorophore for biomolecules labeling such as excellent water solubility, no aggregation in high ionic strength buffer, large extinction coefficient and high fluorescent quantum yield. Antibodies conjugated with IRDye™ 700DX at high D/P ratio exist as monomeric species in high ionic buffer and have bright fluorescence. The IRDye™ 700DX conjugated antibodies generate sensitive, highly specific detection with very low background in Western blot and cytoblot assays.
KW - Bioconjugate
KW - Cytoblot assay
KW - Immunoblotting assay
KW - Isomericly pure
KW - Near-infrared fluorescence
KW - Photobleaching quantum yield
KW - Photostable dye
KW - Water-soluble phthalocyanine dye
UR - http://www.scopus.com/inward/record.url?scp=33646194994&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33646194994&partnerID=8YFLogxK
U2 - 10.1117/12.669173
DO - 10.1117/12.669173
M3 - Conference contribution
AN - SCOPUS:33646194994
SN - 0819461393
SN - 9780819461391
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
ER -