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生物感測器原理與應用. 綠色螢光蛋白質之細胞學應用. Fluorescence Spectra. Wavelength: . c: 3 10 8 m/s 可見光波長： 400 ~ 700 nm 波數（ Wave number) 為波長 的倒數. Principles of Fluorescence. Aromatic molecules: Delocalized electrons in conjugated double bonds. Stokes’ Shift. 1852, Sir G. G. Stokes in Cambridge.
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生物感測器原理與應用 綠色螢光蛋白質之細胞學應用
Fluorescence Spectra Wavelength: c: 3 108 m/s 可見光波長：400 ~ 700 nm 波數（Wave number) 為波長 的倒數
Principles of Fluorescence Aromatic molecules: Delocalized electrons in conjugated double bonds
Stokes’ Shift 1852, Sir G. G. Stokes in Cambridge
Kasha’s Rule 1950 The same fluorescence emission spectrum is generally observed irrespective of the excitation wavelength. The vibrational relaxation ~ 1 ps
Green Fluorescent Protein Jellyfish Aequorea victoria The spontaneous cyclization and oxidation of the sequence – Ser65 (Thr65) – Tyr66 – Gly67 – 11-stranded barrel with a coaxial helix 238 residues, 27 kDa
Spectra of GFP Absorption maxima 395 and 475 nm Ex: 395 nm Em: 508 nm with QE ~ 0.72 – 0.85 QE (Quantum Yield): the ratio of the no. of photons emitted to the no. absorbed.
Green Fluorescent Protein Imaging of Tumour Growth, Metastasis, and Angiogenesis in Mouse Models Robert M. Hoffman The Lancet Oncology, vol. 3, 546 – 556, 2002
Tumour Progression & Metastasis • High specificity • Strong signal • High resolution • Good physiological conditions • No other proteins, substrates, cofactors • Stability ( ~ 24 days ) • Cytoplasmic marker into any cell type • Unaffected by the external environment • Excitation wavelength is quite long ( ~ 490 nm )
