CODEX multiplexed tissue imaging with DNA-conjugated antibodies

寡核苷酸 DNA 计算生物学 抗体 生物 分子生物学 遗传学
作者
Sarah Black,Darci J. Phillips,John W. Hickey,Julia Kennedy‐Darling,Vishal G. Venkataraaman,Nikolay Samusik,Yury Goltsev,Christian M. Schürch,Garry P. Nolan
出处
期刊:Nature Protocols [Nature Portfolio]
卷期号:16 (8): 3802-3835 被引量:441
标识
DOI:10.1038/s41596-021-00556-8
摘要

Advances in multiplexed imaging technologies have drastically improved our ability to characterize healthy and diseased tissues at the single-cell level. Co-detection by indexing (CODEX) relies on DNA-conjugated antibodies and the cyclic addition and removal of complementary fluorescently labeled DNA probes and has been used so far to simultaneously visualize up to 60 markers in situ. CODEX enables a deep view into the single-cell spatial relationships in tissues and is intended to spur discovery in developmental biology, disease and therapeutic design. Herein, we provide optimized protocols for conjugating purified antibodies to DNA oligonucleotides, validating the conjugation by CODEX staining and executing the CODEX multicycle imaging procedure for both formalin-fixed, paraffin-embedded (FFPE) and fresh-frozen tissues. In addition, we describe basic image processing and data analysis procedures. We apply this approach to an FFPE human tonsil multicycle experiment. The hands-on experimental time for antibody conjugation is ~4.5 h, validation of DNA-conjugated antibodies with CODEX staining takes ~6.5 h and preparation for a CODEX multicycle experiment takes ~8 h. The multicycle imaging and data analysis time depends on the tissue size, number of markers in the panel and computational complexity. This protocol describes co-detection by indexing, a highly multiplexed imaging technology that uses DNA-conjugated antibodies to image up to 60 markers in formalin-fixed, paraffin-embedded and fresh-frozen tissues.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
1秒前
2秒前
2秒前
3秒前
Rosie发布了新的文献求助10
5秒前
5秒前
LKL林完成签到,获得积分10
5秒前
5秒前
1111应助科研通管家采纳,获得10
6秒前
Akim应助科研通管家采纳,获得10
6秒前
6秒前
只A不B应助科研通管家采纳,获得10
6秒前
坦率的匪应助科研通管家采纳,获得10
6秒前
6秒前
李爱国应助科研通管家采纳,获得10
6秒前
田様应助科研通管家采纳,获得10
6秒前
ED应助科研通管家采纳,获得10
6秒前
1111应助科研通管家采纳,获得10
6秒前
7秒前
czh应助科研通管家采纳,获得10
7秒前
7秒前
汉堡包应助拖拖沓沓ttt采纳,获得10
7秒前
坦率的匪应助科研通管家采纳,获得10
7秒前
钮卿完成签到,获得积分10
7秒前
所所应助科研通管家采纳,获得10
7秒前
坦率的匪应助科研通管家采纳,获得10
7秒前
如意的刚应助科研通管家采纳,获得10
7秒前
大模型应助科研通管家采纳,获得10
7秒前
7秒前
7秒前
7秒前
7秒前
7秒前
7秒前
7秒前
7秒前
linmo发布了新的文献求助10
8秒前
8秒前
华仔应助那咋了采纳,获得10
8秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3988827
求助须知:如何正确求助?哪些是违规求助? 3531197
关于积分的说明 11252739
捐赠科研通 3269830
什么是DOI,文献DOI怎么找? 1804815
邀请新用户注册赠送积分活动 881915
科研通“疑难数据库(出版商)”最低求助积分说明 809028