生物感測器原理與應用

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

Mirror Image Rule

Intrinsic Biochemical Fluorophores

Extrinsic Fluorophores

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

GFP

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.

Microscope

Filter Set

Spatial Resolution

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 )

GFP in Veins & Capillaries

Metastasis in Brain

Bone Metastasis

Single Cancer Cell

Angiogenesis of Pancreatic Tumour

Lymphoma Dissemination

GFP and Luciferase

生物感測器原理與應用

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