Flow cytometry, in conjunction with modeling algorithms, provides a powerful tool to assess cells in G0/G1 phase versus S phase, G2, or polyploidy. We offer a series of fluorescent dyes to allow accurate cell cycle analysis in either live or fixed cell populations.

Select cell cycle products

See also imaging assays for live-cell cell cycle analysis

Live cell cycle assays

The Invitrogen Vybrant DyeCycle family of dyes offers robust fluorescent dyes for live cell cycle analysis with limited cytotoxicity using 405, 488, 532, or 633 nm excitation. The dyes have low cytotoxicity, allowing stained cells to be sorted and otherwise cultured or assessed with functional assays after staining.

 
Cell cycle analysis using Vybrant DyeCycle Violet stain. Histogram of live Jurkat cells stained with Vybrant DyeCycle Violet stain showing DNA content distribution. G0/G1 and G2/M peaks are separated by the S phase distribution. Violet 405 nm excitation was used with a 440/40 nm bandpass filter.

 

Fixed cell cycle assays

Analyzing cell cycle using Invitrogen FxCycle Violet, Invitrogen SYTOX AADvanced, or Invitrogen FxCycle Far Red stains allows for multiple color options for simplified fixed cell cycle analysis.

 
Multiparametric cell cycle and immunophenotypic analysis. TF-1 erythroblast cells were alcohol-fixed overnight, washed, and then suspended in 0.1% Triton X-100/PBS/1% BSA before staining with anti–histone H3[pS10] purified antibody complexed with Zenon Invitrogen Alexa Fluor  488 Rabbit IgG labeling reagent and FxCycle Violet stain. The pH3 signal (red) identifies cells that are in mitosis.

 

Selection guides for cell cycle analysis products

  Hoechst 33342 Vybrant DyeCycle Violet Vybrant DyeCycle Green Vybrant DyeCycle Orange Vybrant DyeCycle Ruby
Basis of assay Utilize fluorescent dyes to distinguish cells in G0/G1 phase versus S phase, G2, or polyploidy
Readout DNA content analysis in living cells using a is cell permeable dye, which after binding double-stranded DNA, emits a fluorescent signal that is proportional to the DNA mass
Excitation UV or 350 UV or 405 488 488 or 532 488 or 633/5
Emission 461 437 534 563 686
Recommended filter 450/50 440/40 530/30 585/42 695/40
Multiplexable
Sample type The dyes have low cytotoxicity, allowing stained cells to be sorted and otherwise cultured or assessed with functional assays after staining.
Format 10 mL 200 assays 200 assays 200 assays 100 assays 400 assays
Cat. No. H3570 V35003 V35004 V35005 V10309 V10273
Bibliography
  Propidium Iodide ReadyProbes Reagent FxCycle Violet FxCycle PI/RNAse Solution FxCycle Far Red SYTOX AADvanced
Basis of assay Utilize fluorescent dyes to distinguish cells in G0/G1 phase versus S phase, G2, or polyploidy
Readout DNA content analysis using a cell permeable dye, which after binding double-stranded DNA when cells are alive, emits a fluorescent signal that is proportional to the DNA mass even after fixation.
Excitation 488 or 532 405 488 or 532 633 488 or 532
Emission 617 358/461 617 660/20 695/40
Recommended filter 575/24 450/50 572/24 660/20 695/40
Multiplexable
RNase required
 
Included
 
Sample type These stains have been designed for flow cytometric analysis of DNA content in fixed cells.
Format 1 kit 1 kit 100 mL 1 kit 5 x 0.5 mL 0.2 mL
Cat. No. R37108 F10347 F10797 F10348 S10274 S10349
Live cell cycle assays
  Hoechst 33342 Vybrant DyeCycle Violet Vybrant DyeCycle Green Vybrant DyeCycle Orange Vybrant DyeCycle Ruby
Basis of assay Utilize fluorescent dyes to distinguish cells in G0/G1 phase versus S phase, G2, or polyploidy
Readout DNA content analysis in living cells using a is cell permeable dye, which after binding double-stranded DNA, emits a fluorescent signal that is proportional to the DNA mass
Excitation UV or 350 UV or 405 488 488 or 532 488 or 633/5
Emission 461 437 534 563 686
Recommended filter 450/50 440/40 530/30 585/42 695/40
Multiplexable
Sample type The dyes have low cytotoxicity, allowing stained cells to be sorted and otherwise cultured or assessed with functional assays after staining.
Format 10 mL 200 assays 200 assays 200 assays 100 assays 400 assays
Cat. No. H3570 V35003 V35004 V35005 V10309 V10273
Bibliography
Fixed cell cycle assays
  Propidium Iodide ReadyProbes Reagent FxCycle Violet FxCycle PI/RNAse Solution FxCycle Far Red SYTOX AADvanced
Basis of assay Utilize fluorescent dyes to distinguish cells in G0/G1 phase versus S phase, G2, or polyploidy
Readout DNA content analysis using a cell permeable dye, which after binding double-stranded DNA when cells are alive, emits a fluorescent signal that is proportional to the DNA mass even after fixation.
Excitation 488 or 532 405 488 or 532 633 488 or 532
Emission 617 358/461 617 660/20 695/40
Recommended filter 575/24 450/50 572/24 660/20 695/40
Multiplexable
RNase required
 
Included
 
Sample type These stains have been designed for flow cytometric analysis of DNA content in fixed cells.
Format 1 kit 1 kit 100 mL 1 kit 5 x 0.5 mL 0.2 mL
Cat. No. R37108 F10347 F10797 F10348 S10274 S10349

Cell cycle analysis scientific posters

  Poster Presented Year
Non-Cytotoxic Near-IR DNA Stain for Cell Cycle Analysis in Living Cells CYTO 2010
Dual Pulse Labeling of S-Phase Population Using Click Chemistry to Measure Changes In Cell Proliferation ASCB 2009
Dual Pulse Labeling Using a New Thymidine Analog 5-Ethynyl-2’-Deoxyuridine (EdU) to Detect Alterations In S-Phase Progression by Fluorescence Microscopy and Flow Cytometry ASCB 2009
Detection of S-Phase Cell Cycle Progression Using 5-Ethynyl-2’-Deoxyuridine Incorporation With Click Chemistry Probe Discovery 2008
Characterization of DNA Content, Cyclin B1 and Phosphorylated Histone H3 With Direct S-Phase Using EdU Incorporation In Multiparameter Testing of Cell Lines With Cell Cycle Blocking Agents ISAC 2008
Detection of DNA Synthesis by Automated Microscopy and Image Analysis: Comparison of BrdU Method and a New Click Chemistry–Based EdU Method ISAC 2008
Effects of The Thymidine Analogues EdU and BrdU On Cell Viability and Cycle Progression ISAC 2008
Evaluation of Click Chemistry–Based Alternative to BrdU Antibody Labeling in Tissue and Cultured Cells Using Fluorescence Microscopy and Flow Cytometry ASCB 2007

Resources

Support

For Research Use Only. Not for use in diagnostic procedures.