Single-Cell Nucleus Extraction with Cellular Indexing

doi:10.1002/advs.202514883

2025-09-01

Trinh Lam

Ana E. Gomez Martinez

Alison Su

Anna Fomitcheva Khartchenko

Xin Wang

Md Nazibul Islam

Amy E. Herr

Herr, Amy Elizabeth *

DOI：10.1002/advs.202514883

原文链接

查看原文

摘要

En 中文

大规模细胞器分离掩盖了细胞间的异质性，而现有的微流控方法无法可靠地将每个分离的细胞器与其母细胞重新连接，这是多组学读出的关键能力。VacTrap是一种高通量微流控装置，用于从哺乳动物细胞中分离和空间索引单个细胞核。VacTrap装置由三层对齐结构组成：1) 带有活门的Bis凝胶微孔层（BAC-gel），其基底被制造在贯穿孔玻璃载玻片上；2) 用于接收转移细胞核的聚二甲基硅氧烷（PDMS）微孔层；3) 真空 manifold。VacTrap的操作始于使用差异去污剂分离（DDF）进行细胞质裂解，以将完整细胞核释放到Bis凝胶微孔中，同时细胞质蛋白被电泳进入Bis凝胶层。随后加入二硫苏糖醇（DTT）和真空处理，可在3-5分钟内溶解活门，同步将细胞核转移至PDMS微孔中，在80%的活门中实现98%的效率。为验证每个细胞核的细胞质蛋白分离，通过档案Bis凝胶层中的原位免疫探针成功检测到特定蛋白靶标。为验证单个完整细胞核的分离和收集，形态分析证实了核特征的保留。通过将细胞核的空间索引重新关联至原始细胞，VacTrap提供了一种稳健、自动化的细胞制备平台，适用于单细胞多组学应用。

Keyword:

hydrogels

microfluidics

multiomics

organelles

proteomics

single-cell

AI总结

对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。

期刊

Advanced Science

IF:

14.1

论文数:

1.8W

被引数:

11.5W

机构

chan zuckerberg initiative (czi)

学者数:

251

论文数: 74

被引数: 0

University of California System

学者数:

37.7W

论文数: 33.8W

被引数: 6.6K

引用论文

IF0

PREAI

Single-cell RNA sequencing technologies and bioinformatics pipelines 单细胞RNA测序技术和生物信息学管道

EXPERIMENTAL AND MOLECULAR MEDICINE

IF12.9

2018-08-07

1.3K

OAAI

Hwang, Byungjin; Lee, Ji Hyun; Bang, Duhee

An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues 改进的ATAC-seq方案降低了背景信号并使冷冻组织得以分析

Nature Methods

IF0

2017-08-28

OAAI

M Ryan Corces; Alexandro E Trevino; Emily G Hamilton; Peyton G Greenside; Nicholas A Sinnott-Armstrong; Sam Vesuna; Ansuman T Satpathy; Adam J Rubin; Kathleen S Montine; Beijing Wu; Arwa Kathiria; Seung Woo Cho; Maxwell R Mumbach; Ava C Carter; Maya Kasowski; Lisa A Orloff; Viviana I Risca; Anshul Kundaje; Paul A Khavari; Thomas J Montine; William J Greenleaf; Howard Y Chang

Live single cell imaging assays in glass microwells produced by laser-induced deep etching

LAB ON A CHIP

IF5.4

2022-01-01

OAAI

Sandstroem, Niklas; Brandt, Ludwig; Sandoz, Patrick A.; Zambarda, Chiara; Guldevall, Karolin; Schulz-Ruhtenberg, Malte; Roesener, Bernd; Krueger, Robin A.; Oenfelt, Bjorn

Magnetophoretic-based microfluidic device for DNA isolation

Biomicrofluidics

IF0

2014-08-22

OAAI

C. Hale; J. Darabi

Microfluidic extraction and stretching of chromosomal DNA from single cell nuclei for DNA fluorescence in situ hybridization 用于DNA荧光原位杂交的单个细胞核中染色体DNA的微流控提取和拉伸

BIOMEDICAL MICRODEVICES

IF3.3

2012-01-10

OAAI

Wang, Xiaozhu; Takebayashi, Shin-ichiro; Bernardin, Evans; Gilbert, David M.; Chella, Ravindran; Guan, Jingjiao

UV Dose Governs UV-Polymerized Polyacrylamide Hydrogel Modulus

international journal of mining science and technology

IF4.4

2017-01-01

OAAI

Sheth, Saahil; Jain, Era; Karadaghy, Amin; Syed, Sana; Stevenson, Hunter; Zustiak, Silviya P.

BAC-DROP: Rapid Digestion of Proteome Fractionated via Dissolvable Polyacrylamide Gel Electrophoresis and Its Application to Bottom-Up Proteomics Workflow

JOURNAL OF PROTEOME RESEARCH

IF3.6

2020-12-24

PREAI

Takemori, Ayako; Ishizaki, Jun; Nakashima, Kenji; Shibata, Takeshi; Kato, Hidemasa; Kodera, Yoshio; Suzuki, Tetsuro; Hasegawa, Hitoshi; Takemori, Nobuaki

Review on recent advances in the analysis of isolated organelles

ANALYTICA CHIMICA ACTA

IF6

2012-11-01

OAAI

Satori, Chad P.; Kostal, Vratislav; Arriaga, Edgar A.

Enhancing protein disulfide bond cleavage by UV excitation and electron capture dissociation for top-down mass spectrometry

International Journal of Mass Spectrometry

IF0

2015-11-01

OAAI

Piriya Wongkongkathep; Huilin Li; Xing Zhang; Rachel R. Ogorzalek Loo; Ryan R. Julian; Joseph A. Loo

查看更多内容