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首页> 外文会议>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：受叶绿素启发的红色和近红外荧光团。多色流式细胞仪和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 ε(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 [ε(λ_(exc) x Φ_f] times 1% 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：流式细胞术对增加多重性（用于检测多种抗原）和亮度（用于检测稀有实体）的需求需要在相对拥挤的可见光谱区域之外具有光谱不同的荧光的新荧光团（即“颜色”）。流式细胞术荧光团通常必须在生物缀合后在水溶液中起作用，并且理想情况下应表现出许多光物理特征：（ⅰ）吸收强，（ⅱ）斯托克斯位移大，（ⅲ）中等强度（如果不是很强的荧光）和（ⅳ）窄荧光带。长期以来一直追求串联染料以实现大的有效斯托克斯位移，增加的亮度以及更好地控制激发和发射波长。在此，介绍了叶绿素和细菌叶绿素（自然界为光合作用选择的光敏颜料）的引人注目的光物理特征，以用于流式细胞术。叶绿素（或细菌叶绿素）构成固有的串联染料，在较高能量的紫外线（UV）或可见吸收带中激发时，由于红色（或近红外）荧光（由于内部快速转换为最低能态），因此发出红色（或近红外）荧光。合成（细菌）二氢卟酚具有强吸收性（近紫外摩尔吸收系数ε（λ_（exc））〜10〜5 M〜（-1）cm〜（-1）），荧光量子产率适中（Φ_f= 0.05） -0.30），根据合成设计可在-600-900 nm范围内调节窄荧光带（10-25 nm）。 “相对实际亮度”由固有亮度[ε（λ_（exc）xΦ_f]乘以发射滤光片在多色实验中捕获的荧光带的分数的1％表示。进行定量评估，以说明预期的8色面板中此类新型荧光团的实际亮度。

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