荧光寿命测定的现代方法与应用
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In 1845, Herschel observed that an otherwise colourless solution of quinine (quinine absorbs in the UV region) in water emitted a blue colour under certain circumstances.
In the 1920s and 1930s Jabłoński investigated polarized light and fluorescence and was able to show that the transition moments in absorption and emission are two different things.
荧光寿命的概念
荧光是分子吸收能量后其基态电子S0被激发到单线激发
态S1或S2后由第一单线激发态S1回到基态S0时所发生的。
荧光寿命是指分子在单线激发态 S1所平均停留的时间，
或者说处于激发态S1的分子数目衰减到原来的1/e所经历的 时间。
由于荧光现象多发生在纳秒级,这正好是分子运动所发生 的时间尺度,因此利用荧光技术可以“看”到许多复杂的分 子间作用过程,例如超分子体系中分子间的簇集、固液界面 上吸附态高分子的构象重排、蛋白质高级结构的变化等。
时间相关单光子记数法(Time-Correlated Single-Photon
Counting , TCSPC)
TCSPC 是目前主要应用的荧光寿命测定技术， 1975 年由PTI (Photon Technology International) 公司首先商品化。此外,Edinburgh Instruments、 IBH、HORIBA 等公司也在生产基于TCSPC 的时间 分辨荧光光谱仪。
It wasn't until the early 1940s that Albert Coons developed a technique for labeling antibodies with fluorescent dyes, thus giving birth to the field of immunofluorescence.
时间相关单光子记数法(Time-Correlated Single-Photon
Counting , TCSPC)
TCSPC 的工作原理如图所示
电
荧光
信
号
累积电压信号
停止工作
电信号
时间相关单光子记数法(Time-Correlated Single-Photon
Counting , TCSPC)
以光子数对时间作图可得到如图所示的直方图,此 图经过平滑处理得到荧光衰减曲线。
The Development of Fluorescence Microscopes
The first fluorescence microscopes were developed between 1911 and 1913 by German physicists Otto Heimstt and Heinrich Lehmann as a spin-off from the ultraviolet instrument.
百度文库
荧光寿命测定的现代方法
荧光寿命测定的现代方法主要有以下几种，即时间相关 单光子记数法(Time-Correlated Single-Photon Counting ， TCSPC) 、频闪技术(Strobe Techniques)、相调制法 (Phase Modulation Methods) 、条纹相机法(Streak Cameras) 和上转换法(Upcon-version Methods)。
In 1852 Sir G.G. Stokes studied the same compound and found that the emitted light has a longer wavelength than the light absorbed, the so-called Stokes’ shift. ( first described fluorescence )
七彩的荧光
主要内容
• 荧光及荧光分光光度计的发展 • 荧光寿命的基本原理 • 荧光寿命测定的现代方法及数据处理 • 测定荧光寿命的一些应用 • 测定荧光寿命的仪器
The Development of Fluorescence
The emission of light from any substance is called luminescence. Some of the first scientific reports of luminescence appeared in the middle of the 18th century.
By the turn of the twenty-first century, the field of fluorescence microscopy was responsible for a revolution in cell biology, coupling the power of live cell imaging to highly specific multiple labeling of individual organelles and macromolecular complexes with synthetic and genetically encoded fluorescent probes.
时间相关单光子记数法(Time-Correlated Single-Photon
Basic theory of fluorescence Lifetime
The photophysical processes that occur from absorption to emission are often shown in a so-called Jabłoński diagram.
In the 1920s and 1930s Jabłoński investigated polarized light and fluorescence and was able to show that the transition moments in absorption and emission are two different things.
荧光寿命的概念
荧光是分子吸收能量后其基态电子S0被激发到单线激发
态S1或S2后由第一单线激发态S1回到基态S0时所发生的。
荧光寿命是指分子在单线激发态 S1所平均停留的时间，
或者说处于激发态S1的分子数目衰减到原来的1/e所经历的 时间。
由于荧光现象多发生在纳秒级,这正好是分子运动所发生 的时间尺度,因此利用荧光技术可以“看”到许多复杂的分 子间作用过程,例如超分子体系中分子间的簇集、固液界面 上吸附态高分子的构象重排、蛋白质高级结构的变化等。
时间相关单光子记数法(Time-Correlated Single-Photon
Counting , TCSPC)
TCSPC 是目前主要应用的荧光寿命测定技术， 1975 年由PTI (Photon Technology International) 公司首先商品化。此外,Edinburgh Instruments、 IBH、HORIBA 等公司也在生产基于TCSPC 的时间 分辨荧光光谱仪。
It wasn't until the early 1940s that Albert Coons developed a technique for labeling antibodies with fluorescent dyes, thus giving birth to the field of immunofluorescence.
时间相关单光子记数法(Time-Correlated Single-Photon
Counting , TCSPC)
TCSPC 的工作原理如图所示
电
荧光
信
号
累积电压信号
停止工作
电信号
时间相关单光子记数法(Time-Correlated Single-Photon
Counting , TCSPC)
以光子数对时间作图可得到如图所示的直方图,此 图经过平滑处理得到荧光衰减曲线。
The Development of Fluorescence Microscopes
The first fluorescence microscopes were developed between 1911 and 1913 by German physicists Otto Heimstt and Heinrich Lehmann as a spin-off from the ultraviolet instrument.
百度文库
荧光寿命测定的现代方法
荧光寿命测定的现代方法主要有以下几种，即时间相关 单光子记数法(Time-Correlated Single-Photon Counting ， TCSPC) 、频闪技术(Strobe Techniques)、相调制法 (Phase Modulation Methods) 、条纹相机法(Streak Cameras) 和上转换法(Upcon-version Methods)。
In 1852 Sir G.G. Stokes studied the same compound and found that the emitted light has a longer wavelength than the light absorbed, the so-called Stokes’ shift. ( first described fluorescence )
七彩的荧光
主要内容
• 荧光及荧光分光光度计的发展 • 荧光寿命的基本原理 • 荧光寿命测定的现代方法及数据处理 • 测定荧光寿命的一些应用 • 测定荧光寿命的仪器
The Development of Fluorescence
The emission of light from any substance is called luminescence. Some of the first scientific reports of luminescence appeared in the middle of the 18th century.
By the turn of the twenty-first century, the field of fluorescence microscopy was responsible for a revolution in cell biology, coupling the power of live cell imaging to highly specific multiple labeling of individual organelles and macromolecular complexes with synthetic and genetically encoded fluorescent probes.
时间相关单光子记数法(Time-Correlated Single-Photon
Basic theory of fluorescence Lifetime
The photophysical processes that occur from absorption to emission are often shown in a so-called Jabłoński diagram.