Laser Capture Microdissection for Analysis of Single Cells

Author(s):  
Nobuki Nakamura ◽  
Katharina Ruebel ◽  
Long Jin ◽  
Xiang Qian ◽  
Heyu Zhang ◽  
...  
1999 ◽  
Vol 70 (11) ◽  
pp. 4377-4385 ◽  
Author(s):  
Seth R. Goldstein ◽  
Thomas Pohida ◽  
Paul D. Smith ◽  
John I. Peterson ◽  
Ed Wellner ◽  
...  

2017 ◽  
Author(s):  
Joseph W. Foley ◽  
Chunfang Zhu ◽  
Philippe Jolivet ◽  
Shirley X. Zhu ◽  
Peipei Lu ◽  
...  

AbstractRNA sequencing (RNA-seq) is a sensitive and accurate method for quantifying gene expression. Small samples or those whose RNA is degraded, such as formalin-fixed, paraffin-embedded (FFPE) tissue, remain challenging to study with nonspecialized RNA-seq protocols. Here we present a new method, Smart-3SEQ, that accurately quantifies transcript abundance even with small amounts of total RNA and effectively characterizes small samples extracted by laser-capture microdissection (LCM) from FFPE tissue. We also obtain distinct biological profiles from FFPE single cells, which have been impossible to study with previous RNA-seq protocols, and we use these data to identify possible new macrophage phenotypes associated with the tumor microenvironment. We propose Smart-3SEQ as a highly cost-effective method to enable large gene-expression profiling experiments unconstrained by sample size and tissue availability. In particular, Smart-3SEQ’s compatibility with FFPE tissue unlocks an enormous number of archived clinical samples, and combined with LCM it allows unprecedented studies of small cell populations and single cells isolated by their in situ context.


2021 ◽  
Author(s):  
Margareth Nogueira ◽  
Daiane CF Golbert ◽  
Richardson Leão

Laser Capture Microdissection (LCM) is a method that allows to select and dissecting specific structures, cell populations, or even single cells from different types of tissue to extract DNA, RNA, or proteins. It is easy to perform and precise, avoiding unwanted signals from irrelevant cells, because gene expression may be affected by a bulk of heterogeneous material in a sample. However, despite its efficiency, several steps can affect the sample RNA integrity. In comparison to DNA, RNA is a much more unstable molecule and represents a challenge in the LCM method. Here we describe an optimized protocol to provide good concentration and high-quality RNA in specific structures, such as Dentate Gyrus and CA1 in the hippocampus, basolateral amygdala and anterior cingulate cortex of mouse brain tissue.


BioTechniques ◽  
1999 ◽  
Vol 26 (2) ◽  
pp. 328-335 ◽  
Author(s):  
C.A. Suarez-Quian ◽  
S.R. Goldstein ◽  
T. Pohida ◽  
P.D. Smith ◽  
J.I. Peterson ◽  
...  

2019 ◽  
Vol 29 (11) ◽  
pp. 1816-1825 ◽  
Author(s):  
Joseph W. Foley ◽  
Chunfang Zhu ◽  
Philippe Jolivet ◽  
Shirley X. Zhu ◽  
Peipei Lu ◽  
...  

2016 ◽  
Vol 77 (S 02) ◽  
Author(s):  
Youssef Yakkioui ◽  
Remco Santegoeds ◽  
Koo van Overbeeke ◽  
Andreas Herrler ◽  
Yasin Temel

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