Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16-30 synchronized fractions, including particle size analysis, can be achieved in 4-5 h.

Original languageEnglish
Pages (from-to)663-673
Number of pages11
JournalNature Protocols
Volume3
Issue number4
DOIs
Publication statusPublished - Apr 2008

Fingerprint

Cell Nucleus
Cell Cycle
Synchronization
Animals
Cells
B-Lymphoid Precursor Cells
Cell Separation
Cricetulus
Cell Size
Particle Size
Ovary
Flow Cytometry
DNA
Flow cytometry
Sedimentation
Particle size analysis

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation. / Bánfalvi, G.

In: Nature Protocols, Vol. 3, No. 4, 04.2008, p. 663-673.

Research output: Contribution to journalArticle

@article{9bd7a027333e4c859d71d5d6bfa74ca8,
title = "Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation",
abstract = "Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16-30 synchronized fractions, including particle size analysis, can be achieved in 4-5 h.",
author = "G. B{\'a}nfalvi",
year = "2008",
month = "4",
doi = "10.1038/nprot.2008.34",
language = "English",
volume = "3",
pages = "663--673",
journal = "Nature Protocols",
issn = "1754-2189",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation

AU - Bánfalvi, G.

PY - 2008/4

Y1 - 2008/4

N2 - Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16-30 synchronized fractions, including particle size analysis, can be achieved in 4-5 h.

AB - Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16-30 synchronized fractions, including particle size analysis, can be achieved in 4-5 h.

UR - http://www.scopus.com/inward/record.url?scp=42149133709&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42149133709&partnerID=8YFLogxK

U2 - 10.1038/nprot.2008.34

DO - 10.1038/nprot.2008.34

M3 - Article

VL - 3

SP - 663

EP - 673

JO - Nature Protocols

JF - Nature Protocols

SN - 1754-2189

IS - 4

ER -