Drop-on-demand single cell isolation and total RNA analysis.
Technologies that rapidly isolate viable single cells from heterogeneous solutions have significantly contributed to the field of medical genomics. Challenges remain both to enable efficient extraction, isolation and patterning of single cells from heterogeneous solutions as well as to keep them ali...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2011-03-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC3055874?pdf=render |
id |
doaj-f0c4d02ea93546f7b709dede8d1046ce |
---|---|
record_format |
Article |
spelling |
doaj-f0c4d02ea93546f7b709dede8d1046ce2020-11-25T01:21:22ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-03-0163e1745510.1371/journal.pone.0017455Drop-on-demand single cell isolation and total RNA analysis.Sangjun MoonYun-Gon KimLingsheng DongMichael LombardiEdward HaeggstromRoderick V JensenLi-Li HsiaoUtkan DemirciTechnologies that rapidly isolate viable single cells from heterogeneous solutions have significantly contributed to the field of medical genomics. Challenges remain both to enable efficient extraction, isolation and patterning of single cells from heterogeneous solutions as well as to keep them alive during the process due to a limited degree of control over single cell manipulation. Here, we present a microdroplet based method to isolate and pattern single cells from heterogeneous cell suspensions (10% target cell mixture), preserve viability of the extracted cells (97.0±0.8%), and obtain genomic information from isolated cells compared to the non-patterned controls. The cell encapsulation process is both experimentally and theoretically analyzed. Using the isolated cells, we identified 11 stem cell markers among 1000 genes and compare to the controls. This automated platform enabling high-throughput cell manipulation for subsequent genomic analysis employs fewer handling steps compared to existing methods.http://europepmc.org/articles/PMC3055874?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sangjun Moon Yun-Gon Kim Lingsheng Dong Michael Lombardi Edward Haeggstrom Roderick V Jensen Li-Li Hsiao Utkan Demirci |
spellingShingle |
Sangjun Moon Yun-Gon Kim Lingsheng Dong Michael Lombardi Edward Haeggstrom Roderick V Jensen Li-Li Hsiao Utkan Demirci Drop-on-demand single cell isolation and total RNA analysis. PLoS ONE |
author_facet |
Sangjun Moon Yun-Gon Kim Lingsheng Dong Michael Lombardi Edward Haeggstrom Roderick V Jensen Li-Li Hsiao Utkan Demirci |
author_sort |
Sangjun Moon |
title |
Drop-on-demand single cell isolation and total RNA analysis. |
title_short |
Drop-on-demand single cell isolation and total RNA analysis. |
title_full |
Drop-on-demand single cell isolation and total RNA analysis. |
title_fullStr |
Drop-on-demand single cell isolation and total RNA analysis. |
title_full_unstemmed |
Drop-on-demand single cell isolation and total RNA analysis. |
title_sort |
drop-on-demand single cell isolation and total rna analysis. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2011-03-01 |
description |
Technologies that rapidly isolate viable single cells from heterogeneous solutions have significantly contributed to the field of medical genomics. Challenges remain both to enable efficient extraction, isolation and patterning of single cells from heterogeneous solutions as well as to keep them alive during the process due to a limited degree of control over single cell manipulation. Here, we present a microdroplet based method to isolate and pattern single cells from heterogeneous cell suspensions (10% target cell mixture), preserve viability of the extracted cells (97.0±0.8%), and obtain genomic information from isolated cells compared to the non-patterned controls. The cell encapsulation process is both experimentally and theoretically analyzed. Using the isolated cells, we identified 11 stem cell markers among 1000 genes and compare to the controls. This automated platform enabling high-throughput cell manipulation for subsequent genomic analysis employs fewer handling steps compared to existing methods. |
url |
http://europepmc.org/articles/PMC3055874?pdf=render |
work_keys_str_mv |
AT sangjunmoon dropondemandsinglecellisolationandtotalrnaanalysis AT yungonkim dropondemandsinglecellisolationandtotalrnaanalysis AT lingshengdong dropondemandsinglecellisolationandtotalrnaanalysis AT michaellombardi dropondemandsinglecellisolationandtotalrnaanalysis AT edwardhaeggstrom dropondemandsinglecellisolationandtotalrnaanalysis AT roderickvjensen dropondemandsinglecellisolationandtotalrnaanalysis AT lilihsiao dropondemandsinglecellisolationandtotalrnaanalysis AT utkandemirci dropondemandsinglecellisolationandtotalrnaanalysis |
_version_ |
1725130693931958272 |