Measuring photosynthetic parameters in individual algal cells by Fast Repetition Rate fluorometry
Measuring photosynthetic parameters in individual algal cells by Fast Repetition Rate fluorometry
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DOI：
10.1023/a:1006360005033
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发表时间：
1999-12-01
影响因子：
3.7
通讯作者：
Falkowski, PG
中科院分区：
文献类型：
--
作者：
Gorbunov, MY;Kolber, ZS;Falkowski, PG
研制了单细胞快速重复率（SCFRR）荧光计，用于测量单个藻细胞光化学量子产率、PS Ⅱ功能吸收截面和PS Ⅱ受体侧电子传递动力学。这些参数被用来量化的细胞特定的光合性能在水生生态系统中的天然浮游植物的珊瑚礁。SCFRR技术测量叶绿素荧光瞬变引起的一系列精确控制的激发闪光灯，累积饱和PS II在120 μ s内。为了满足对单个藻类细胞的分析要求，测量在微体积中进行，使得一次探测多于一个细胞的概率非常低。我们设计了一种新颖的、计算机控制的液压机械系统，将一部分样品输送到测量室中，测量后，将其转移到六个分选容器中的一个中。的荧光信号是由一系列的高频率闪光灯从高亮度的蓝色发光二极管，并获得了一种新的红色敏感的光电倍增管为基础的检测系统，表现出高灵敏度和非常宽的动态范围。宽动态范围的检测器允许SCFRR测量各种各样的细胞大小范围从1到100 μ m的等效球直径。紧凑和重量轻的设计使SCFRR荧光计适用于实验室和现场研究。
A Single Cell Fast Repetition Rate (SCFRR) fluorometer was developed to measure the quantum yield of photochemistry, the functional absorption cross section of PS II and the kinetics of electron transport on the acceptor side of PS II in individual algal cells. These parameters are used to quantify the cell-specific photosynthetic performance in natural phytoplankton assembledges in aquatic ecosystems. The SCFRR technique measures chlorophyll fluorescence transients induced by a precisely controlled series of excitation flashlets that cumulatively saturate PS II within 120 mu s. To meet the requirement in the analysis for single algal cells, the measurements are conducted in micro volumes, such that the probability of probing more than one cell at a time is vanishingly low. We designed a novel, computer-controlled hydromechanical system to deliver a portion of the sample into the measuring chamber and, following measurement, remove it into one of six sorting containers. The fluorescence signal is induced by a series of high frequency flashlets obtained from high luminosity blue light-emitting diodes and is acquired by a novel red-sensitive PMT-based detection system exhibiting both high sensitivity and a very wide dynamic range. The wide dynamic range of the detector allows SCFRR measurements for a wide variety of cell sizes ranging from 1 to 100 mu m equivalent spherical diameter. The compact and light-weight design makes the SCFRR Fluorometer applicable for both laboratory and field studies.