Improved pre-treatment for lipid staining of microalgae with a rigid cell wall

IF 1.9 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2025-07-25 DOI:10.1016/j.mimet.2025.107205

Samira Reuscher , Gerd Klock , Antje Kersten , Samuel Schabel , Rüdiger Graf

{"title":"Improved pre-treatment for lipid staining of microalgae with a rigid cell wall","authors":"Samira Reuscher , Gerd Klock , Antje Kersten , Samuel Schabel , Rüdiger Graf","doi":"10.1016/j.mimet.2025.107205","DOIUrl":null,"url":null,"abstract":"<div><div>The worldwide depletion of energy and feedstock resources is an important topic of the current decade. Microalgae are an interesting option, offering rapid biomass production in combination with CO<sub>2</sub>-fixation. For ongoing research in microalgae applications, well-functioning laboratory methods are important. One major aspect in microalgae-related studies is the measurement of the biomass' lipid content. In order to avoid elaborate extraction protocols, lipid staining with fluorescent dyes such as Nile Red or BODIPY is currently preferred in small-scale screening trials. Although practical, staining was hindered for microalgae with a rigid cell wall. Additional pre-treatment using solvents has already been described, but this was not sufficient for all strains. Besides sonication, we show here that fluorescent staining can be successfully implemented following the disruption of microalgal cell walls by bead beating. The protocol functions with conventional laboratory equipment (ball mill, shaker and vortex mixer), all of them allowing for small-volume samples and processing of multiple samples in parallel. The bead beating method extends the fluorescence lipid staining approach to microalgae with an extremely rigid cell wall and can be applied for screening trials in biofuel or biomaterial related research.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"236 ","pages":"Article 107205"},"PeriodicalIF":1.9000,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of microbiological methods","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0167701225001216","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 0

Abstract

The worldwide depletion of energy and feedstock resources is an important topic of the current decade. Microalgae are an interesting option, offering rapid biomass production in combination with CO₂-fixation. For ongoing research in microalgae applications, well-functioning laboratory methods are important. One major aspect in microalgae-related studies is the measurement of the biomass' lipid content. In order to avoid elaborate extraction protocols, lipid staining with fluorescent dyes such as Nile Red or BODIPY is currently preferred in small-scale screening trials. Although practical, staining was hindered for microalgae with a rigid cell wall. Additional pre-treatment using solvents has already been described, but this was not sufficient for all strains. Besides sonication, we show here that fluorescent staining can be successfully implemented following the disruption of microalgal cell walls by bead beating. The protocol functions with conventional laboratory equipment (ball mill, shaker and vortex mixer), all of them allowing for small-volume samples and processing of multiple samples in parallel. The bead beating method extends the fluorescence lipid staining approach to microalgae with an extremely rigid cell wall and can be applied for screening trials in biofuel or biomaterial related research.

Abstract Image

查看原文本刊更多论文

具有刚性细胞壁的微藻脂质染色预处理方法的改进

世界范围内能源和原料资源的枯竭是近十年来的一个重要话题。微藻是一个有趣的选择，它能提供快速的生物质生产和二氧化碳固定。对于正在进行的微藻应用研究，功能良好的实验室方法非常重要。微藻相关研究的一个主要方面是生物量脂质含量的测量。为了避免复杂的提取方案，目前在小规模筛选试验中首选使用荧光染料（如尼罗河红或BODIPY）进行脂质染色。虽然可行，但对于细胞壁坚硬的微藻，染色受到阻碍。已经描述了使用溶剂的额外预处理，但这并不足以适用于所有菌株。除了超声，我们在这里表明，荧光染色可以成功地实施微藻细胞壁的破坏后，头跳动。该协议与传统的实验室设备（球磨机，激振器和涡旋混合器）一起使用，所有这些设备都允许小批量样品和并行处理多个样品。该方法将荧光脂质染色方法扩展到细胞壁极其坚硬的微藻，可用于生物燃料或生物材料相关研究的筛选试验。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.