Tissue-Like 3D Standard and Protocols for Microscope Quality Management

Author:

Abrams Benjamin1ORCID,Pengo Thomas2ORCID,Wee Tse-Luen34ORCID,Deagle Rebecca C34ORCID,Vuillemin Nelly34,Callahan Linda M5ORCID,Smith Megan A3,Kubow Kristopher E6ORCID,Girard Anne-Marie7,Rappoport Joshua Z8,Bayles Carol J9,Cameron Lisa A10ORCID,Cole Richard11,Brown Claire M34ORCID

Affiliation:

1. Life Sciences Microscopy Center, University of California, Santa Cruz , 1156 High Street, 150 Sinsheimer Labs, Santa Cruz, CA 95064 , USA

2. Informatics Institute, University of Minnesota Twin Cities, Cancer and Cardiovascular Research Building , 2231 6th St SE, Minneapolis, MN 55449 , USA

3. Advanced BioImaging Facility (ABIF), McGill University , 3649 Prom, Sir William Osler, Bellini Building, Room 137, Montreal, QC H3G 0B1 , Canada

4. Department of Physiology, McGill University , 3655 Promenade Sir-William-Osler, Montreal, Quebec H3G 1Y6 , Canada

5. Department of Neuroscience, Del Monte Institute for Neuroscience, Univ. Rochester Medical Center , Rochester, NY 14642 , USA

6. Biology Department, James Madison University , Bioscience Building, 951 Carrier Drive, Harrisonburg, VA 22807 , USA

7. Center for Genome Research and Biocomputing, Oregon State University , 1500 SW Jefferson Way, Corvallis, OR 97331 , USA

8. Center for Advanced Microscopy and Nikon Imaging Center, Feinberg School of Medicine, Northwestern Medicine, Northwestern University , Chicago, IL 60611 , USA

9. Institute of Biotechnology, Cornell University , Cornell Institute of Biotechnology, Microscopy Facility, 130 Biotechnology Building, Cornell University Ithaca, NY 14853 , USA

10. Light Microscopy Core Facility, Duke University , 4215 French Family Science Center, 124 Science Drive, Durham, NC 27708 , USA

11. New York State Dept of Health/Wadsworth Center, Advanced Light Microscopy & Image Analysis Core Facility , 150 New Scotland Ave, Albany, NY 12208 , USA

Abstract

AbstractThis article outlines a global study conducted by the Association of Biomedical Resource Facilities (ABRF) Light Microscopy Research Group (LMRG). The results present a novel 3D tissue-like biologically relevant standard sample that is affordable and straightforward to prepare. Detailed sample preparation, instrument-specific image acquisition protocols and image analysis methods are presented and made available to the community. The standard consists of sub-resolution and large well characterized relative intensity fluorescence microspheres embedded in a 120 µm thick 3D gel with a refractive index of 1.365. The standard allows the evaluation of several properties as a function of depth. These include the following: 1) microscope resolution with automated analysis of the point-spread function (PSF), 2) automated signal-to-noise ratio analysis, 3) calibration and correction of fluorescence intensity loss, and 4) quantitative relative intensity. Results demonstrate expected refractive index mismatch dependent losses in intensity and resolution with depth, but the relative intensities of different objects at similar depths are maintained. This is a robust standard showing reproducible results across laboratories, microscope manufacturers and objective lens types (e.g., magnification, immersion medium). Thus, these tools will be valuable for the global community to benchmark fluorescence microscopes and will contribute to improved scientific rigor and reproducibility.

Publisher

Oxford University Press (OUP)

Subject

Instrumentation

Cited by 1 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3