The cell cycle is a series of growth and development steps which a cell undergoes during cell division. Comprised of a set of distinct phases, the cell cycle is a key drug discovery target and of significant importance for both cancer and regenerative medicine research. Indeed a dis-regulation of the cell cycle is a hallmark of cancer, and check point inhibitors and cyclin dependent kinases that regulate cell cycle are studied as strategies or targets for anti-cancer therapies. Cell cycle regulation is also believed to play a key role in the cell differentiation that is required to deliver on the promise of regenerative medicine. However, studying dynamic changes of the cell cycle can be challenging.
Conventional methods to study the cell cycle may be challenging, due to:
Continuous imaging and analysis over time at microplate throughput, within a physiologically-relevant environment inside your incubator, enables the long-term, kinetic analysis of cell cycle dynamics. With the quantification of cell cycle progression within living cells, researchers can:
With the Incucyte® Cell Cycle Assay, you can now quantify cell-cycle progression continuously over multiple cell divisions - inside your incubator. Gain deeper insight into treatment effects on cell cycle dynamics, or link cell cycle arrest to morphology and function using our unique and accessible approach to live-cell imaging and analysis.
The Incucyte Cell Cycle Assay combines easy-to-learn automated image acquisition and integrated analysis of 96- or 384-well plates with a simple, non-perturbing protocol for cell cycle labeling to generate kinetic, image-based measurements of cell cycle progression over time – all from inside your incubator. The Incucyte® Cell Cycle Lentivirus Reagents are fluorescent ubiquitination-based cell cycle indicator (FUCCI), and provide a homogeneous expression of two fluorescent proteins to distinguish between cells in the interphase or the mitotic phase to enable the visualization of cell cycle progression. The Incucyte® Cell Cycle Green/Red Lentivirus Reagent and the Incucyte® Cell Cycle Green/Orange Lentivirus Reagent are both available for use in the Cell Cycle Assay. Used in combination with the Incucyte® Cell-by-Cell Analysis Software Module, the Cell Cycle Assay offers a powerful solution for tracking subpopulations of cells within each stage of the cell cycle over multiple cell divisions, enabling greater insight into cell cycle dynamics.
Follow cell cycle progression over time in living cells — inside your incubator
Combine a simple protocol with easy-to-learn automated image acquisition and integrated analysis of 96- or 384-well plates
Enable high-throughput study of drug-induced treatment effects on cell cycle transitions or link cell cycle modulation to function
Follow cell cycle progression over multiple cell divisions in a variety of cell types using image-based measurements. Maintain physiological relevance with stable incubation provided by your incubator and our novel Incucyte Cell Cycle Lentivirus Reagents that are non-perturbing to cell health and morphology.
Figure 1. Generate image-based measurements of cell cycle transitions over multiple cell divisions with automated image acquisition and integrated analysis. HeLa cells infected with the Incucyte Cell Cycle Green/Red Lentivirus Reagent exhibit red fluorescence in G1 and green fluorescence in S/G2/M (A). Yellow cells are in transition from G1 to S while non-fluorescent cells are moving from M to G1. Quantification using Incucyte Cell-by-Cell Analysis enables accurate masking shown by yellow mask (B) and classification based on red and green fluorescence (C&D). Panel E shows quantification of cell cycle duration, where HeLa cells transfected with Incucyte Cell Cycle Green/Red Lentivirus Reagent were treated with Thymidine (2.5 mM for 24 h) to arrest cell growth in S/G2/M phase. Upon release of this block, cells undergo synchronous division (M-G1-S/G2/M). Data shows a cycle length of 17 h (peak to peak duration). Data shown as mean ± SEM of 6 wells.
Figure 2. Generate direct measurements within living cells through non-perturbing reagents. The introduction of the Incucyte Green/Red Cell Cycle Lentivirus Reagent stably into cells is non-perturbing. Images show the same morphology in parent or transfected MDA-MB-231 cells and analysis of images demonstrates no effect on cell growth.
Speed time to answer with Incucyte's approachable, end-to-end solution. Follow a simple live-cell labelling protocol - no fixing, no lifting. Automate acquisition and analysis in 96- or 384-well plates with easy-to-use, purpose-built software tools. Easily generate graphs to reveal kinetic changes in cell cycle phase distribution using Incucyte Cell-by-Cell Analysis Software Module.
Figure 3. Incucyte Cell Cycle Lentivirus Reagent Quick Guide. Stably express a fluorescent ubiquitinated-cell cycle indicator (FUCCI) using a simple labelling protocol — no fixing, no lifting.
Combine the Incucyte Cell Cycle Lentivirus Reagent with Cell-by-Cell Analysis to track individually identified phases of the cell cycle for the evaluation of drug treatment effects, or link drug-induced cell cycle arrest to cell differentiation.
Figure 4. Measure drug effects on cell cycle phase distribution over time. Time course of HT1080 fibrosarcoma cell division following cisplatin or fluorouracil (5FU) treatment. Cell cycle phases were determined in HT1080 cells expressing the Incucyte Cell Cycle Green/Red Lentivirus Reagent. Images were analysed using Cell-by-Cell Analysis software and population subsets classified based on green and red fluorescence. Microplate views (top row) show the concentration and time dependent effects over 30 h of cisplatin and 5FU compared to vehicle for the G1 phase (red population), S/G2/M phase (green population) and G1-S phase (orange population). The concentration response curves taken at 24 h post treatment (bottom row) show after cisplatin treatment, there was a decrease in the population of cells in the G1 phase, increase in the S/G2/M phase and decrease in the G-S phase in line with its effect to interfere with mitosis. After 5FU treatment, there was an increase in the population of cells in the G1 phase, decrease in the S/G2/M and G1-S phase in line with effect on DNA synthesis. Values shown are the mean ± SEM of 6 wells.
Figure 5. Observe drug-induced cell cycle arrest effects on cell morphology and function. THP-1 monocytes stably expressing the Incucyte Cell Cycle Green/Red Lentivirus Reagent were exposed to vehicle or Phorbol 12-myristate 13-acetate (PMA; 100 nM) and analysed using the Cell-by-Cell Analysis software. PMA causes the cells to differentiate arresting the cell cycle in the G1 phase (A). Analysis demonstrates a lack of proliferation (B), increase percentage of red cells, G1 phase and a decrease in green cells S/G2/M phase (C) compared to vehicle treated cells. PMA-treated cells were also larger in area (D), indicative of a morphological change and concordant increase in phagocytic potential as measured by efferocytosis of apoptotic Jurkat cells labeled with the Incucyte pHrodo Red Cell Labeling Kit (E).