建立结晶紫吸光度与生物膜生物量之间的定量联系

IF 1.9 Q2 MULTIDISCIPLINARY SCIENCES
MethodsX Pub Date : 2025-09-16 DOI:10.1016/j.mex.2025.103630
Sheida Stephens , Radhakrishnan Mahadevan , D. Grant Allen
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引用次数: 0

摘要

量化生物量对于研究生物制造中的生物膜至关重要,然而广泛使用的测定方法,如结晶紫(CV),往往产生难以跨实验室比较的结果,因为它依赖于吸光度作为生物量的主观代表。为了解决这个问题,我们提出了一种标准化的方法,使用离心浮游生物培养物校准CV吸光度与细胞光密度(OD)和干细胞重量(DCW)。通过建立OD、DCW和CV吸光度之间的三向相关性,这种简单的方法可以进行定量、可重复性和可比性的生物量测量。在大肠杆菌菌株和palustr红假单胞菌之间的验证显示出很强的线性,特别是当使用10%醋酸作为溶剂时。评估了季节性和基于仪器的变异性,支持该方法的稳健性和广泛的实用性。该方法使研究人员能够标准化CV数据,提高实验室间的可比性,并支持更准确的生物膜生产力评估。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Establishing a quantitative link between crystal violet absorbance and biomass in biofilms

Establishing a quantitative link between crystal violet absorbance and biomass in biofilms
Quantifying biomass is essential for studying biofilms in biomanufacturing, yet widely used assays such as crystal violet (CV) often yield results that are difficult to compare across laboratories due to its reliance on absorbance as a subjective proxy for biomass. To address this, we present a standardized method that calibrates CV absorbance against cellular optical density (OD) and dry cell weight (DCW) using centrifuged planktonic cultures. By establishing a three-way correlation among OD, DCW, and CV absorbance, this simple method allows for quantitative, reproducible, and comparable biomass measurements. Validation across Escherichia coli strains and Rhodopseudomonas palustris showed strong linearity, particularly when using 10% acetic acid as the solvent. Seasonal and instrument-based variability were evaluated, supporting the method’s robustness and broad utility. This method enables researchers to normalize CV data, improving inter-laboratory comparability and supporting more accurate assessment of biofilm productivity.
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来源期刊
MethodsX
MethodsX Health Professions-Medical Laboratory Technology
CiteScore
3.60
自引率
5.30%
发文量
314
审稿时长
7 weeks
期刊介绍:
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