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首页> 外文会议>SPIE Conference on Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications >Red and near-infrared fluorophores inspired by chlorophylls. Consideration of practical brightness in multicolor flow cytometry and biomedical sciences
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Red and near-infrared fluorophores inspired by chlorophylls. Consideration of practical brightness in multicolor flow cytometry and biomedical sciences

机译：红色和近红外荧光团受到叶绿素的启发。多色流式细胞术与生物医学中实际亮度的考虑

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Demands in flow cytometry for increased multiplexing (for detection of multiple antigens) and brightness (for detection of rare entities) require new fluorophores (i.e., "colors") with spectrally distinct fluorescence outside the relatively congested visible spectral region. Flow cytometry fluorophores typically must function in aqueous solution upon bioconjugation and ideally should exhibit a host of photophysical features: (i) strong absorption, (ii) sizable Stokes shift, (iii) modest if not strong fluorescence, and (iv) narrow fluorescence band. Tandem dyes have long been pursued to achieve a large effective Stokes shift, increased brightness, and better control over the excitation and emission wavelengths. Here, the attractive photophysical features of chlorophylls and bacteriochlorophylls - Nature's chosen photoactive pigments for photosynthesis - are described with regards to use in flow cytometry. A chlorophyll (or bacteriochlorophyll) constitutes an intrinsic tandem dye given the red (or near-infrared) fluorescence upon excitation in the higher energy ultraviolet (UV) or visible absorption bands (due to rapid internal conversion to the lowest energy state). Synthetic (bacterio)chlorins are available with strong absorption (near-UV molar absorption coefficient e(λ_(exc)) ~10~5 M~(-1) cm~(-1)), modest fluorescence quantum yield (Φ_f= 0.05-0.30), and narrow fluorescence band (10-25 nm) tunable from ~600-900 nm depending on synthetic design. The "relative practical brightness" is given by intrinsic brightness [e(λ_(exc)) x Φ_f] times η_f, the fraction of the fluorescence band that is captured by an emission filter in a multicolor experiment. The spectroscopic features of (bacterio)chlorins are evaluated quantitatively to illustrate practical brightness for this novel class of fluorophores in a prospective 8-color panel.
机译：流式细胞术中的需求增加复用（用于检测多次抗原）和亮度（用于检测稀有实体）需要具有谱不同荧光的新荧光团（即“颜色”）在相对拥挤的可见光谱区域之外的光谱明显荧光。流式细胞术荧光团通常必须在生物缀合物上用水溶液起作用，理想地应表现出一系列的光学特征：（i）强烈的吸收，（ii）可大幅度的斯托克斯转移，（iii）适度的荧光，（iv）窄荧光带。长期以来一直追求串联染料，以实现大型有效的斯托克斯转移，增加亮度，更好地控制激发和发射波长。在此，关于在流式细胞术中使用，描述了用于光合作用的叶绿素和菌肤氯苯基 - 自然选择的光学活性色素的吸引力。叶绿素（或Bacteriochorrophyl1）构成在较高能量紫外线（UV）或可见吸收带中的激发时给予的固有串联染料（由于快速内部转化为最低能量状态），赋予红色（或近红外）荧光。合成（细菌）氯有很强的吸收（接近UV摩尔吸收系数E（λ_（Exc））〜10〜5 m〜（-1）cm〜（-1）），荧光量子产量适度（φ_f= 0.05 -0.30），窄荧光带（10-25nm）可根据合成设计从〜600-900nm调谐。 “相对实际亮度”由本征亮度[e（λ_（ex））xφ_f]倍η_f，在多色实验中被发射滤波器捕获的荧光带的分数。 （细菌）氯共的光谱特征定量评价，以说明在预期8色面板中的这种新型荧光团的实际亮度。

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