荧光光谱 荧光光谱法特点： 灵敏度高、选择性强、用样少、方法简便以及能提供较多的物理参数等优点。用途广泛， 如可用于研究溶液中蛋白质构象。

Presentation on theme: "荧光光谱 荧光光谱法特点： 灵敏度高、选择性强、用样少、方法简便以及能提供较多的物理参数等优点。用途广泛， 如可用于研究溶液中蛋白质构象。"— Presentation transcript:

1 荧光光谱 荧光光谱法特点： 灵敏度高、选择性强、用样少、方法简便以及能提供较多的物理参数等优点。用途广泛， 如可用于研究溶液中蛋白质构象。

2 By Felix A. Carroll, Perspectives on Structure and Mechanism In Organic chemistry
Brooks/Cole Publishing Company, 1998

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5 KASHA RULE Polyatomic molecular entities luminesce with appreciable yield only from the lowest excited state of a given multiplicity. There are exceptions to this rule. KASHA-VAVILOV RULE The quantum yield of luminescence is independent of the wavelength of exciting radiation. There are exceptions to this rule.

6 Anomalous Fluorescence Patterns in Corals
We have noticed several kinds of anomalous patterns in corals that show up clearly in fluorescence but are not visible under white-light illumination. These include: bright or dark patches within areas of more uniform fluorescence isolated patches of fluorescent color color patterns that are only seen in fluorescence The significance of these effects is completely unknown, but they almost certainly point to underlying differences in internal chemistry.

7 White-light (left) and fluorescence (right) photographs of a coral, Roatan, Honduras. Note the yellow-fluorescent area in the photograph on the right that is not evident in the other image.

8 2。基本知识： （－〕. 激发光谱与发射光谱

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14 荧光熄灭剂：引起荧光熄灭的物质。如：卤素离子， 重金属离子， 硝基化合物， O2等。
荧光光谱法是研究水溶液中蛋白质分子构象的一种新方法：通过测定蛋白质分子的自身荧光； 另一是向蛋白质的特殊部位，引入荧光探测剂， 然后测定荧光探测剂的荧光。

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17 向蛋白质分子引入荧光探测剂，运用荧光偏振法，可以测定：蛋白质分子的聚合与解离；抗原与抗体的反应机制；不同蛋白质分子相互作用等。

18 日立

19 激发光 光源--激发单色器 比色杯--透射光 | 荧光 荧光单色器 探测器(计算机) 荧光光度计示意图

20 DNA: Fluorescence? NO. In experiment: DNA + EB high Fluo.
Mechanism: Intercalation (嵌入)

21 Circadian Rhythms in Ethidium Bromide (He, Y. J. et al
Circadian Rhythms in Ethidium Bromide (He, Y.J. et al., Medical Hypotheses 2000, 54(5): )

22 Mechanism: Intercalation (嵌入)

23 Circadian Rhythms in RNase A (Left, a: 30 C; b: 37 C) and DNA (Poly(dA*dT)-EB) (He, Y.J. et al., Medical Hypotheses 2000, 55(3): ) Fluorescence (A.u.) Time (hr) in Washington DC, 12:00 (Noon) is starting point

24 Protein: Fluorescence?
在蛋白质中，能发射荧光的氨基酸残基，只有Try (348 nm), Tyr (303 nm)和Phe (282 nm)。个别蛋白质含有黄素腺嘌呤二核苷酸 (FAD)，也能了射荧光。

25 芳香族类 (全部共三种氨基酸)

26 Circadian Rhythms in RNase A (Left, a: 30 C; b: 37 C) and DNA (Poly(dA*dT)-EB) (He, Y.J. et al., Medical Hypotheses 2000, 55(3): ) Fluorescence (A.u.) Time (hr) in Washington DC, 12:00 (Noon) is starting point

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28 实验内容 目的: 理解荧光产生的机理，学会荧光物质激发与发射波长的确定，测定温度效应等． 1. 溴乙锭的激发光谱测定
目的:　理解荧光产生的机理，学会荧光物质激发与发射波长的确定，测定温度效应等． 1. 溴乙锭的激发光谱测定 2. 溴乙锭的发射光谱测定 3. 在固定激发与发射波长下, 温度对溴乙锭的荧光强度的影响

29 实验报告的基本写法 １．标题 ２．姓名，单位．．． ３．摘要 ４．前言（为什么做此实验） ５．材料与方法 ６．结果与讨论 ７．结论
８．参考文献

30 THE END THANKS!