流式细胞术和光学显微镜定量评估细胞活力的比较

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该文章用钙黄绿素AM和EthD-1作为染料，比较了用流式细胞术和光镜来定量评估细胞活力，发现两者效果一致。

染料介绍：

• Calcein AM is a cell-permeant dye that can be used to determine cell viability in most eukaryotic cells. In live cells the nonfluorescent calcein AM is converted to a green-fluorescent calcein after acetoxymethyl ester hydrolysis by intracellular esterases.
• The cell-impermeant viability indicator ethdium homodimer-1 (EthD-1) is a high-affinity nucleic acid stain that is weakly fluorescent until bound to DNA and emits red fluorescence
  

文章摘要：
Cytometry A. 2012 Oct 18. doi: 10.1002/cyto.a.22213. [Epub ahead of print]  
Quantitative assessment of cell viability based on flow cytometry and microscopy.
Kummrow A   , Frankowski M   , Bock N   , Werner C   , Dziekan T   , Neukammer J   .
Physikalisch-Technische Bundesanstalt, Berlin, Germany. andreas.kummrow@ptb.de.

Abstract
We compare flow cytometric and microscopic determination of cell viability by fluorescence labeling using calcein acetoxy-methyl-ester and ethidium homodimer-1 as live and dead stain, respectively. Peripheral blood monocytes served as model system and were accumulated applying density gradients. Subsequently, monocytes were further enriched by magnetic-activated or fluorescence-activated cell sorting (MACS, FACS) targeting the antigen CD14. Identical samples were used for flow cytometric and microscopic analysis to allow direct comparison of both analysis methods. More than 1,000 cells were measured for each sample to minimize the measurement uncertainty caused by counting statistics. We observed good agreement of flow cytometric and microscopic viability measurements. On average, the difference in viability measured by flow cytometry and microscopy amounted to (2.7 ± 1.4)% for live staining and (1.7 ± 1.2)% for dead staining. These deviations were similar to the uncertainty of measurement for cell viability, thus demonstrating that both methods delivered equal results. Besides monocytes, comparison of flow cytometric and microscopy viability for MACS enriched CD34-positive cells also showed consistent results. © 2012 International Society for Advancement of Cytometry.

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