Xu Manlong, McCanna David J, Sivak Jacob G
Optometry and Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
Centre for Contact Lens Research, Optometry and Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
J Pharmacol Toxicol Methods. 2015 Jan-Feb;71:1-7. doi: 10.1016/j.vascn.2014.11.003. Epub 2014 Nov 15.
PrestoBlue is a new resazurin based reagent to assess cell viability and cytotoxicity. It is claimed to be a fast and highly sensitive assay. Here, we compared PrestoBlue, alamarBlue, and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazonium bromide (MTT) in assessing cell viability of human corneal epithelial cells (HCEC), and investigated the effect of plate color, reading mode, and plate storage on the performance of PrestoBlue assay.
The viability of different numbers of healthy HCEC and the toxicity of various chemicals on HCEC were evaluated using PrestoBlue (fluorescence), alamarBlue (fluorescence), and MTT (absorbance). The sensitivities of the three assays were compared. In the PrestoBlue assay, three plate colors and two reading modes were used and compared in assessing the toxic effect of sodium dodecyl sulfate (SDS). The PrestoBlue solutions after reaction were stored and measured on day 1, 2, 3, 5, and 7. The fluorescence readings obtained on different days were then compared.
Both PrestoBlue and alamarBlue were able to detect 5000 healthy cells after 30min incubation and 1000 cells after 1h, 2h, and 4h incubation; while MTT was able to detect 5000 cells after 3h incubation. In the assessment of the toxicity of various chemicals, PrestoBlue and alamarBlue performed similarly. There was no significant difference between the results obtained by these two reagents. All the three plate colors and two reading modes showed similar results in the PrestoBlue assay in assessing the toxicity of SDS. Plate storage up to 7days did not affect the result of the PrestoBlue assay.
Our study suggests that in evaluating the viability of HCEC, PrestoBlue is more sensitive than MTT, but similar to alamarBlue. The plate color, reading mode and plate storage up to 7days did not affect the performance of the PrestoBlue assay.
PrestoBlue是一种基于刃天青的新型试剂，用于评估细胞活力和细胞毒性。据称它是一种快速且高度灵敏的检测方法。在此，我们比较了PrestoBlue、alamarBlue和3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑（MTT）在评估人角膜上皮细胞（HCEC）活力方面的效果，并研究了板的颜色、读数模式和板的储存对PrestoBlue检测性能的影响。
使用PrestoBlue（荧光法）、alamarBlue（荧光法）和MTT（吸光度法）评估不同数量健康HCEC的活力以及各种化学物质对HCEC的毒性。比较这三种检测方法的灵敏度。在PrestoBlue检测中，使用三种板的颜色和两种读数模式来比较评估十二烷基硫酸钠（SDS）的毒性作用。反应后的PrestoBlue溶液在第1、2、3、5和7天进行储存和测量。然后比较不同日期获得的荧光读数。
孵育30分钟后，PrestoBlue和alamarBlue均能够检测到5000个健康细胞，孵育1小时、2小时和4小时后能够检测到1000个细胞；而MTT在孵育3小时后能够检测到5000个细胞。在评估各种化学物质的毒性时，PrestoBlue和alamarBlue表现相似。这两种试剂获得的结果之间没有显著差异。在PrestoBlue检测中评估SDS毒性时，所有三种板的颜色和两种读数模式都显示出相似的结果。板储存长达7天不影响PrestoBlue检测的结果。
我们的研究表明，在评估HCEC的活力时，PrestoBlue比MTT更灵敏，但与alamarBlue相似。板的颜色、读数模式以及板储存长达7天均不影响PrestoBlue检测的性能。
