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Advanced Flow Cytometry with 3-Color, Coulter Volume and Side Scatter Analysis
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Developed from a unique combination of ingenuity and high-level technical resources, the Cell Lab Quanta SC is a flow cytometry system that simultaneously measures electronic volume, side scatter and three fluorescent colors to provide unsurpassed population resolution and accurate cell counting. This advanced flow cytometer has multiple excitation wavelengths, including 488 nm laser and a UV light source optimized for excitation at 366, 405, 435 nm. Together with interchangeable filters, the Quanta SC allows flexible fluorochrome selection for a variety of multicolor applications typically only achievable with expensive and complicated systems. Side scatter and Coulter volume make size analysis and fluorescent measurements more precise. Unique user features expand ease-of-use and flexibility. When all the details are added together, the sum is performance.
Now Available - Increase Throughput with the Quanta SC MPL (Multi-Platform Loader) Option
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More Info
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Application ExamplesViability
Cell impermeant dyes, such as propidium iodide (PI) or 7-Amino Actinomycin D (7-AAD), can be used to differentiate live and dead cells. Because of loss in plasma membrane integrity, dead cells are stained positive (left panel). Differences in volume between live and dead cells can also be measured. Dead cells can be further identified with a dual parameter measurement of volume and side scatter (right panel).
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Three color immunophenotyping
Concentration and population measurements utilizing CD markers can be performed on the Quanta SC. Lymphocytes that are stained with CD4-FITC, CD8-PE and CD3-PC5 emit green, orange and red fluorescence when excited with the 488 nm laser.
 
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Apoptosis
Stages of programmed cell death can be enumerated using a dual parameter staining consisting of Annexin V-FITC and 7-AAD. Phosphatidylserine (PS) resides on the inner cell membrane of healthy cells, but will externalize to the outer membrane layer when in early stage apoptosis. Annexin V-FITC binds to the exposed PS molecules and can be measured. Cells with damaged plasma membrane are stained 7-AAD. Early apoptotic cells are Annexin V-FITC positive but 7-AAD negative and late apoptotic/dead cells are both Annexin V-FITC and 7-AAD positive (Q2).
Control |
| Control |
| Region |
Cell Population |
% Total |
Annexin-V (-),
7-AAD (+) |
Dead cells |
0.55 |
Annexin-V (+),
7-AAD (+) |
Late apoptotic/dead cells |
5.43 |
Annexin-V (-),
7-AAD (-) |
Live cells |
90.46 |
Annexin-V (+),
7-AAD (-) |
Early apoptotic cells |
3.56 |
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Treated |
| Treated |
| Region |
Cell Population |
% Total |
Annexin-V (-),
7-AAD (+) |
Dead cells |
1.97 |
Annexin-V (+),
7-AAD (+) |
Late apoptotic/dead cells |
22.58 |
Annexin-V (-),
7-AAD (-) |
Live cells |
64.01 |
Annexin-V (+),
7-AAD (-) |
Early apoptotic cells |
11.04 |
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Active Caspase-3
Active caspase-3 in apoptotic cells is detected with anti-Cleaved Caspase-3 (Asp175) – Alexa Fluor 488 and analyzed with the Quanta SC.
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CONTROL
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Pct |
Color |
| Caspase 3- |
96.55% |
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| Caspase 3+ |
3.45% |
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TREATED
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Pct |
Color |
| Caspase 3- |
86.67% |
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| Caspase 3+ |
13.33% |
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CFP expression
CFP expression can be detected using the UV light source of the Quanta SC (422/39 exciter and a 480/30 BP filter). The exciter includes both 405 nm and 435 nm of the UV light source. Expression of CFP stable transfected yeast was identified with these excitation and emission settings (top panel) compared to the control (bottom panel).
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Cell cycle analysis using UV
Enumeration of the various phases of a cell is easily determined using a DNA stain such as DAPI or Hoechst. The Quanta SC utilizes a Mercury arc lamp that produces an ideal 366 nm excitation line. G0/G1, S and G2/M population can be measured using fluorescence and contrasted against cell volume.
| Region |
Diameter (µm) |
MCV
(µm3) |
Count |
Conc.
(per ml x 1000) |
Percentage of Total |
FL1 Mean |
FL1 HPCV |
FL1 CV |
Color |
| G0/G1 |
6.22 |
125.7 |
10,901 |
304.2 |
62.21% |
191.4 |
2.78% |
4.86% |
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| S |
7.06 |
183.9 |
4,313 |
120.3 |
24.61% |
265.3 |
6.07% |
11.71% |
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| G2/M |
7.58 |
228.4 |
1,664 |
46.5 |
9.50% |
351.1 |
4.30% |
3.79% |
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Two color cell cycle
Cyclin A2-FITC and 7-AAD can be used together to further interrogate the various G1, S, G2 and M phases of the cell cycle. Cyclin A2 is expressed at very low level in G0 and G1 cells. Its expression gradually increases as cells enter the S phase. Maximal expression of cyclin A2 is found in G2 cells. Measurement of cyclin A2 expression can be used to distinguish mitotic cells (cyclin A2 negative) from G2 cells (cyclin A2 positive).
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Bacterial viability and enumeration
Count live and dead bacterial populations using a double color staining consisting of Syto9 and PI. Syto9, a cell permeable dye, binds to DNA of all the cells. PI stains only cells with damaged or compromised membrane integrity. Concentration and percentage of live and dead cells are calculated with accuracy.
100% dead (treated with 70% ethanol) |
50% dead |
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Cell proliferation
Carbosyfluorescein diacetate, succinimidyl ester (CFSE) is a cell permeable dye that enters cells, covalently binds to intracellular amines and yields fluorescence after cleaved by intracellular esterases. Once the cell divides the dye remains in the two daughter cells at a lower concentration. A cell stained with CFSE can be kept in culture for several days. Each fluorescent peak represents another round of cell division. The area of each peak is the number of cells in that particular cycle.
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