Cell assay using a two-photon-excited europium chelate
| Autor: | Gregory W. Faris, Jeanne P. Haushalter, Kenneth T. Kotz, Xudong Xiao |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
ocis:(180.4315) Nonlinear microscopy
ocis:(170.5810) Scanning microscopy Microscope chemistry.chemical_element ocis:(190.7220) Upconversion 010402 general chemistry Photochemistry 01 natural sciences law.invention 010309 optics Optics Two-photon excitation microscopy law 0103 physical sciences Fluorescence microscope Microscopy business.industry Chemistry ocis:(170.2520) Fluorescence microscopy Photobleaching Atomic and Molecular Physics and Optics Photon upconversion 0104 chemical sciences Autofluorescence Epidermoid carcinoma ocis:(170.1530) Cell analysis business Europium Biotechnology |
| Zdroj: | Biomedical Optics Express |
| ISSN: | 2156-7085 |
| Popis: | We report application of two-photon excitation of europium chelates to immunolabeling of epidermal growth factor receptor (EGFR) cell surface proteins on A431 cancer cells. The europium chelates are excited with two photons of infrared light and emit in the visible. Europium chelates are conjugated to antibodies for EGFR. A431 (human epidermoid carcinoma) cells are labeled with this conjugate and imaged using a multiphoton microscope. To minimize signal loss due to the relatively long-lived Eu(3+) emission, the multiphoton microscope is used with scanning laser two-photon excitation and non-scanning detection with a CCD. The chelate labels show very little photobleaching (less than 1% during continuous illumination in the microscope for 20 minutes) and low levels of autofluorescence (less than 1% of the signal from labeled cells). The detection limit of the europium label in the cell assay is better than 100 zeptomoles. |
| Databáze: | OpenAIRE |
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