Download NC-3000™ Cell Cycle Assays

NC-3000™ Cell Cycle Assays
– For rapid measurement of G1/G0, S and G2/M cell cycle phases
The cell cycle represents the most fundamental and important process in eukaryotic cells and is an ordered set of events, culminating in cell growth and division into two daughter cells.
In a given population, cells will be distributed among three major
phases of the cell cycle: G1 /G0 phase (one set of paired chromosomes per cell), S phase (DNA synthesis with variable amount of
DNA), and G2/M phase (two sets of paired chromosomes per cell,
prior to cell division).
The NucleoCounter® NC-3000™ offers two different cell cycle assays with predefined settings: 1) the rapid and simple Two-step
Cell Cycle Assay with a 5 minutes protocol, and 2) the robust Cell
Cycle Assay of Fixed Cells, offering the possibility of storing cells to
be analyzed for several weeks.
Two-step Cell Cycle
STEP 1: lysis buffer + DAPI
5 min. - detachment, disaggregation
and staining of cells
Key Benefits
of the NC-3000™ Cell Cycle Assays
Perform a complete cell cycle
analysis in just 5 minutes!
 Fast and easy measurement of cell cycle phases at the single cell level
 Acquisition, analysis and data
presentation in one simple step
 Standardized results - even with
different users
 No RNase treatment required
Cell Cycle of Fixed Cells
 No calibration required
Ethanol fixation of cells
STEP 2: stabilization buffer
 PlotManager for superior data
presentation
 Automated PDF reports
Wash with PBS
0 min. - neutralization of sample
 Export of data in FCS/ACS formats
Image Cytometry
Stain cells with DAPI
5 MINUTES
Image Cytometry
PLE
ROBUST & SIM
PROTOCOL
N,
ACQUISITIO
ANALYSIS AND
TATION
DATA PRESEN
IN ONE
SIMPLE STEP
The NucleoCounter® NC-3000™
- Next generation cell analysis
FIXED
ASSAYS
HIGH SPEED
CELL COUNT
FAST
ANALYSIS
VISUAL
INSPECTION
NO
RINSING
NO
CLOGGING
NO
CALIBRATION
NO
MAINTENANCE
LEARN
MORE
Viability and Cell Count | Cell Cycle Assays | Caspase 3/7, 8 & 9 | Annexin V | Mitochondrial Potential | Cell Vitality | DNA Fragmentation | GFP Transfection Efficiency | FlexiCyte™ (user adaptable protocols)
Principle: NC-3000™ Cell Cycle Assays
Using fluorescence microscopy and image analysis, the NucleoCounter® NC-3000™ system automates DNA content quantification and hence, measurements of cell cycle stages. The NC-3000™
Cell Cycle Assays use the nuclear stain, DAPI, to measure DNA content. DAPI binds specifically to
double-stranded DNA and, accordingly, there is no requirement for removing RNA prior to DNA
content measurements. This is a prerequisite for other dyes commonly used for measurements of
cell cycle phases, such as propidium iodide.
The Cell Cycle
After sample preparation cells are loaded into either of two types of ChemoMetec slides: the
2-chamber NC-Slide A2™ or the 8-chamber NC-Slide A8™. Samples are analyzed using the
NucleoCounter® NC-3000™ system and cellular fluorescence is quantified and DNA content histograms are displayed on PC screen. Markers in the displayed histograms can be used to demarcate
cells in the different cells cycle stages.
NC-3000™ Two-step Cell Cycle Assay
The Two-Step Cell Cycle Assay facilitates detaching, permeabilization, declumping and homogenous staining of the cell population in two simple steps without any trypsination, washing and
centrifugation. Preparation of samples only takes 5 minutes.
NC-3000™ Cell Cycle of Fixed Cells Assay
The Cell Cycle of Fixed Cells Assay possesses the possiblity of storing cells for several weeks. Thus,
after ethanol fixation the cells can be left in the fridge for a longer period before analysing the
sample.
Stages of the cell cycle can be estimated by
DNA content measurements. The genome is
replicated during S-phase (seen as a gradual increase in DNA content). In contrast,
cells in G1 and G2/M are uniform with
respect to DNA content, which is equivalent
to the DNA ploidy index 1.0 (for G1) and 2.0
(for G2/M).
990-0302 VER 01.2014
Results: Presented in PlotManager
U2OS cells were grown in the absence (upper row) or in the presence (lower row) of etoposide and the DNA content was measured using
the Two-Step Cell Cycle Assay and a NucleoCounter ® NC-3000™. Scatter plots, histograms and tableswere obtained from the NucleoView™
NC-3000™ software. Markers in the displayed histograms were used to demarcate cells in the different cells cycle phases. Colored histogram
is a merge between untreated (blue line) and etoposide treated (red line) samples.
For more information, please visit
ChemoMetec A/S
Gydevang 43
DK-3450 Allerod
Denmark
www.chemometec.com/NC-3000
Phone(+45) 48 13 10 20
Fax (+45) 48 13 10 21
[email protected]
Webwww.chemometec.com
www.youtube.com/chemometec