FLIMB: fluorescence lifetime microendoscopy for metabolic and functional imaging of femoral marrow at subcellular resolution.

IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Biomedical optics express Pub Date : 2025-03-31 eCollection Date: 2025-04-01 DOI:10.1364/BOE.549311
Alexander F Fiedler, Ruth Leben, Herbert Stürmer, Robert Günther, Romano Matthys, Reto Nützi, Anja E Hauser, Raluca A Niesner
{"title":"FLIMB: fluorescence lifetime microendoscopy for metabolic and functional imaging of femoral marrow at subcellular resolution.","authors":"Alexander F Fiedler, Ruth Leben, Herbert Stürmer, Robert Günther, Romano Matthys, Reto Nützi, Anja E Hauser, Raluca A Niesner","doi":"10.1364/BOE.549311","DOIUrl":null,"url":null,"abstract":"<p><p>Intravital imaging of bone marrow provides a unique opportunity to study cellular dynamics and their interaction with the tissue microenvironment, which governs cell functions and metabolic profiles. To optically access the deep marrow of long bones, we previously developed a microendoscopy system for longitudinal two-photon fluorescence imaging of the murine femur. However, this does not provide information on cell functions or metabolism, for which quantification fluorescence lifetime imaging (FLIM) has proven to be a versatile tool. We present and characterize FLIMB, an adapted GRIN-based microendoscopic system capable of performing reliable, co-registered TCSPC-based two-photon excited FLIM and fluorescence imaging in the femur of fluorescent reporter mice, at sub-cellular resolution. Using FLIMB, we demonstrate metabolic imaging via NAD(P)H-FLIM and intracellular Ca<sup>2+</sup> signaling via FRET-FLIM in immune cell subsets, in the femoral marrow. This method retains the power to study molecular mechanisms underlying various cell functions in tissue context thus providing new insights into bone biology.</p>","PeriodicalId":8969,"journal":{"name":"Biomedical optics express","volume":"16 4","pages":"1711-1731"},"PeriodicalIF":2.9000,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12047734/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomedical optics express","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1364/BOE.549311","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/4/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

Abstract

Intravital imaging of bone marrow provides a unique opportunity to study cellular dynamics and their interaction with the tissue microenvironment, which governs cell functions and metabolic profiles. To optically access the deep marrow of long bones, we previously developed a microendoscopy system for longitudinal two-photon fluorescence imaging of the murine femur. However, this does not provide information on cell functions or metabolism, for which quantification fluorescence lifetime imaging (FLIM) has proven to be a versatile tool. We present and characterize FLIMB, an adapted GRIN-based microendoscopic system capable of performing reliable, co-registered TCSPC-based two-photon excited FLIM and fluorescence imaging in the femur of fluorescent reporter mice, at sub-cellular resolution. Using FLIMB, we demonstrate metabolic imaging via NAD(P)H-FLIM and intracellular Ca2+ signaling via FRET-FLIM in immune cell subsets, in the femoral marrow. This method retains the power to study molecular mechanisms underlying various cell functions in tissue context thus providing new insights into bone biology.

荧光寿命显微内窥镜用于亚细胞分辨率的股骨骨髓代谢和功能成像。
骨髓活体成像为研究细胞动力学及其与组织微环境的相互作用提供了一个独特的机会,组织微环境控制着细胞功能和代谢谱。为了光学地进入长骨的深层骨髓,我们先前开发了一种用于小鼠股骨纵向双光子荧光成像的显微内窥镜系统。然而,这并不能提供细胞功能或代谢的信息,而定量荧光寿命成像(FLIM)已被证明是一种通用的工具。我们提出并描述了flam,这是一种基于grin的显微内镜系统,能够在荧光报告小鼠的股骨中以亚细胞分辨率进行可靠的、共同注册的基于tcpc的双光子激发FLIM和荧光成像。使用FLIMB,我们通过NAD(P)H-FLIM和细胞内Ca2+信号通过FRET-FLIM在股骨髓免疫细胞亚群中进行代谢成像。这种方法保留了在组织背景下研究各种细胞功能的分子机制的能力,从而为骨生物学提供了新的见解。
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
Biomedical optics express
Biomedical optics express BIOCHEMICAL RESEARCH METHODS-OPTICS
CiteScore
6.80
自引率
11.80%
发文量
633
审稿时长
1 months
期刊介绍: The journal''s scope encompasses fundamental research, technology development, biomedical studies and clinical applications. BOEx focuses on the leading edge topics in the field, including: Tissue optics and spectroscopy Novel microscopies Optical coherence tomography Diffuse and fluorescence tomography Photoacoustic and multimodal imaging Molecular imaging and therapies Nanophotonic biosensing Optical biophysics/photobiology Microfluidic optical devices Vision research.
×
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