利用拉曼显微光谱技术单细胞测量微生物的生长速度。

IF 3.5 3区 生物学 Q2 MICROBIOLOGY
Tristan A Caro, Srishti Kashyap, George Brown, Claudia Chen, Sebastian H Kopf, Alexis S Templeton
{"title":"利用拉曼显微光谱技术单细胞测量微生物的生长速度。","authors":"Tristan A Caro, Srishti Kashyap, George Brown, Claudia Chen, Sebastian H Kopf, Alexis S Templeton","doi":"10.1093/femsec/fiae110","DOIUrl":null,"url":null,"abstract":"<p><p>Rates of microbial growth are fundamental to understanding environmental geochemistry and ecology. However, measuring the heterogeneity of microbial activity at the single-cell level, especially within complex populations and environmental matrices, remains a forefront challenge. Stable isotope probing (SIP) is a method for assessing microbial growth and involves measuring the incorporation of an isotopic label into microbial biomass. Here, we assess Raman microspectroscopy as a SIP technique, specifically focusing on the measurement of deuterium (2H), a tracer of microbial biomass production. We correlatively measured cells grown in varying concentrations of deuterated water with both Raman spectroscopy and nanoscale secondary ion mass spectrometry (nanoSIMS), generating isotopic calibrations of microbial 2H. Relative to Raman, we find that nanoSIMS measurements of 2H are subject to substantial dilution due to rapid exchange of H during sample washing. We apply our Raman-derived calibration to a numerical model of microbial growth, explicitly parameterizing the factors controlling growth rate quantification and demonstrating that Raman-SIP can sensitively measure the growth of microorganisms with doubling times ranging from hours to years. The measurement of single-cell growth with Raman spectroscopy, a rapid, nondestructive technique, represents an important step toward application of single-cell analysis into complex sample matrices or cellular assemblages.</p>","PeriodicalId":12312,"journal":{"name":"FEMS microbiology ecology","volume":" ","pages":""},"PeriodicalIF":3.5000,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11347945/pdf/","citationCount":"0","resultStr":"{\"title\":\"Single-cell measurement of microbial growth rate with Raman microspectroscopy.\",\"authors\":\"Tristan A Caro, Srishti Kashyap, George Brown, Claudia Chen, Sebastian H Kopf, Alexis S Templeton\",\"doi\":\"10.1093/femsec/fiae110\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Rates of microbial growth are fundamental to understanding environmental geochemistry and ecology. However, measuring the heterogeneity of microbial activity at the single-cell level, especially within complex populations and environmental matrices, remains a forefront challenge. Stable isotope probing (SIP) is a method for assessing microbial growth and involves measuring the incorporation of an isotopic label into microbial biomass. Here, we assess Raman microspectroscopy as a SIP technique, specifically focusing on the measurement of deuterium (2H), a tracer of microbial biomass production. We correlatively measured cells grown in varying concentrations of deuterated water with both Raman spectroscopy and nanoscale secondary ion mass spectrometry (nanoSIMS), generating isotopic calibrations of microbial 2H. Relative to Raman, we find that nanoSIMS measurements of 2H are subject to substantial dilution due to rapid exchange of H during sample washing. We apply our Raman-derived calibration to a numerical model of microbial growth, explicitly parameterizing the factors controlling growth rate quantification and demonstrating that Raman-SIP can sensitively measure the growth of microorganisms with doubling times ranging from hours to years. The measurement of single-cell growth with Raman spectroscopy, a rapid, nondestructive technique, represents an important step toward application of single-cell analysis into complex sample matrices or cellular assemblages.</p>\",\"PeriodicalId\":12312,\"journal\":{\"name\":\"FEMS microbiology ecology\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2024-08-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11347945/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"FEMS microbiology ecology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1093/femsec/fiae110\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"FEMS microbiology ecology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/femsec/fiae110","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

微生物的生长速率是了解环境地球化学和生态学的基础。然而,在单细胞水平上测量微生物活动的异质性,尤其是在复杂的种群和环境基质中测量微生物活动的异质性,仍然是一项前沿挑战。稳定同位素探测(SIP)是一种评估微生物生长的方法,涉及测量微生物生物量中同位素标签的结合情况。在这里,我们将拉曼微光谱技术作为一种 SIP 技术进行评估,特别是侧重于氘(2H)的测量,氘是微生物生物量产生的示踪剂。我们利用拉曼光谱和纳米级二次离子质谱(nanoSIMS)对生长在不同浓度氘化水中的细胞进行了相关测量,从而得出微生物 2H 的同位素定标。与拉曼光谱法相比,我们发现纳米级二次离子质谱法测量的 2H 会因样品洗涤过程中 H 的快速交换而被大量稀释。我们将源自拉曼的校准应用于微生物生长的数值模型,对控制生长率量化的因素进行了明确的参数化,并证明拉曼-SIP 可以灵敏地测量倍增时间从数小时到数年不等的微生物生长。拉曼光谱是一种快速、无损的技术,它对单细胞生长的测量是将单细胞分析应用于复杂样品基质或细胞组合的重要一步。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Single-cell measurement of microbial growth rate with Raman microspectroscopy.

Rates of microbial growth are fundamental to understanding environmental geochemistry and ecology. However, measuring the heterogeneity of microbial activity at the single-cell level, especially within complex populations and environmental matrices, remains a forefront challenge. Stable isotope probing (SIP) is a method for assessing microbial growth and involves measuring the incorporation of an isotopic label into microbial biomass. Here, we assess Raman microspectroscopy as a SIP technique, specifically focusing on the measurement of deuterium (2H), a tracer of microbial biomass production. We correlatively measured cells grown in varying concentrations of deuterated water with both Raman spectroscopy and nanoscale secondary ion mass spectrometry (nanoSIMS), generating isotopic calibrations of microbial 2H. Relative to Raman, we find that nanoSIMS measurements of 2H are subject to substantial dilution due to rapid exchange of H during sample washing. We apply our Raman-derived calibration to a numerical model of microbial growth, explicitly parameterizing the factors controlling growth rate quantification and demonstrating that Raman-SIP can sensitively measure the growth of microorganisms with doubling times ranging from hours to years. The measurement of single-cell growth with Raman spectroscopy, a rapid, nondestructive technique, represents an important step toward application of single-cell analysis into complex sample matrices or cellular assemblages.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
FEMS microbiology ecology
FEMS microbiology ecology 生物-微生物学
CiteScore
7.50
自引率
2.40%
发文量
132
审稿时长
3 months
期刊介绍: FEMS Microbiology Ecology aims to ensure efficient publication of high-quality papers that are original and provide a significant contribution to the understanding of microbial ecology. The journal contains Research Articles and MiniReviews on fundamental aspects of the ecology of microorganisms in natural soil, aquatic and atmospheric habitats, including extreme environments, and in artificial or managed environments. Research papers on pure cultures and in the areas of plant pathology and medical, food or veterinary microbiology will be published where they provide valuable generic information on microbial ecology. Papers can deal with culturable and non-culturable forms of any type of microorganism: bacteria, archaea, filamentous fungi, yeasts, protozoa, cyanobacteria, algae or viruses. In addition, the journal will publish Perspectives, Current Opinion and Controversy Articles, Commentaries and Letters to the Editor on topical issues in microbial ecology. - Application of ecological theory to microbial ecology - Interactions and signalling between microorganisms and with plants and animals - Interactions between microorganisms and their physicochemical enviornment - Microbial aspects of biogeochemical cycles and processes - Microbial community ecology - Phylogenetic and functional diversity of microbial communities - Evolutionary biology of microorganisms
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信