Chemotaxis Cell Migration & Invasion
Cell migration is a fundamental biological process and a critical component of human development, immune response and diseases such as tumor metastasis. Under normal, healthy conditions, the capacity for cells to migrate is tightly regulated – e.g., the directed cell migration (chemotaxis) of leukocytes in response to chemokine release from damaged tissue, which plays a key role in the inflammatory response. In contrast, unregulated cell migration and invasion is a hallmark of diseases such as cancer, where metastatic cells move away from the primary tumor and establish secondary tumors at other sites in the organism.
Measurement of chemotaxis in cells is an essential component for understanding disease progression and immune response. However, there are drawbacks to current analysis methods. Common limitations of existing chemotaxis cell-based assays, such as trasnwell, include:
- Inability to visualize cell morphology of migration or invasion in real time
- Low throughput with cumbersome protocols requiring fixing, staining or cell scraping steps
- Requirement of a large number of cells
- Lack of integrated quantitation for measuring kinetic cell movement
Incucyte® Chemotaxis Cell Migration and Invasion Assays are image-based solutions, consisting of optically clear Incucyte® Clearview 96-well Plates and integrated software for continuous visualization and reproducible analysis of active cell motility. Monitor every cell in your experiment like never before!
Solutions for Chemotaxis
Incucyte® Chemotaxis Cell Migration & Invasion Assay Overview
Track and quantify dynamic cell movement using the Incucyte® Clearview 96-well Plate for Chemotaxis. Our optically clear surface enables non-invasive visualization and label-free analysis of cell movement. Incucyte® Clearview 96-well Plates have been validated for use with Incucyte® Live-Cell Analysis Systems and require the Incucyte® Chemotaxis Analysis Software Module.
Figure 1. Incucyte® Clearview 96-Well Plate for Chemotaxis. The Incucyte® Clearview 96-well Plate is specially designed to optimize chemotaxis assays and contains 96 laser-etched pores with a diameter of 8 µm. Cells are added to the upper chamber and chemoattractant to the lower reservoir plate. The design of the Incucyte® Clearview 96-well Plate ensures a stable chemotactic gradient over 72 hours and requires only 1,000-5,000 cells per well, ideal for experiments requiring precious, rare samples. Chemotaxis is visually monitored while cells remain in the physiologically relevant environment of the incubator.
Incucyte® Chemotaxis Analysis Software Module
Analyze label-free and fluorescently labeled chemotactic cell migration or invasion while qualitatively monitoring cell morphology of every cell within each well of a Incucyte® 96-well Clearview Plates.
Cat. No. 9600-0015
Live-Cell Imaging and Analysis by Sartorius
The Incucyte® Live-Cell Analysis System allows you to visualize and quantify label-free or fluorescently labeled chemotactic cell migration, invasion and transendothelial migration in real time—directly inside your incubator. Both time-lapse imaging and quantitative data are automatically generated during incubation, providing powerful insights into the time-course of chemotaxis in a physiologically relevant environment. This instrument, in conjunction with the Incucyte® Clearview 96-well Plates and Incucyte® Chemotaxis Analysis Software Module, offers a powerful alternative to end-point measurements and a better understanding of chemotaxis.
Key Advantages of Incucyte® Chemotaxis Cell Migration & Invasion Assay
Quantify and visualize cell migration or invasion of every cell in your experiment in real time with fully automated analysis
See example data below
Use Incucyte® Clearview Plates to conduct highly reproducible 96-well kinetic assays suitable for screening
See example data below
Reduce cell requirement with optimized pore density and stable gradient, ideal for rare or expensive cells
See example data below
Example Data for Chemotaxis Cell Migration & Invasion Assay
Visualize cell migration in real time with fully automated analysis
Monitor every cell in your experiment, assess morphology and gain phenotypic insight from images and movies using phase contrast and/or fluorescent imaging.
Movie 1. Real-time quantitative analysis of HT-1080 chemotaxis. Chemotactic migration of HT-1080 cells toward FBS, visualized on an Incucyte® Clearview 96-well Plate. Integrated Incucyte® Chemotaxis Analysis Software allows for real-time quantification of cells as they move through pores and adhere to the bottom side of the membrane.
Watch more movies:
Reduce cell requirement with optimized pore density and stable gradient
Figure 2. Lower cell requirement using the Incucyte® Chemotaxis Assay. Cell usage comparison for the Incucyte® Clearview 96-well Plate and a modified Boyden-chamber. Jurkat T cell chemotaxis toward SDF-1α was measured in both systems (2,000 to 20,000 cells/well). However, no signal was observed with the modified Boyden-plate. A minimum of 150,000 cells/well was required to observe chemotactic migration using the Boyden system.
Figure 3. Evaluation of Incucyte® Chemotaxis Assay gradient. (A) 10,000 kDA dextran, labeled with Alexa Fluor® 594 was added to the reservoir plates of an Incucyte® Clearview 96-well Plate and a 96-well modified Boyden-chamber at a concentration of 10 µM to establish a gradient. Gradient stability was measured at 0, 4,8, 24, 48 and 72 hours (N=3 wells). After 4 hours, a rapid loss in gradient is measure in the modified Boyden-chamber, whereas the Incucyte® Clearview Plate maintains a linear gradient over 72 hours. (B) An FBS gradient was established over 72, 48, 24 and t=0hours in separate wells. Serum starved Incucyte® Nuclight Red HT-1080 cells were added to the established gradients and FBS-induced chemotactic response was measured. Each data point represents mean ± SEM, N=3.
Investigate treatment effects on migration and invasion
Screen treatments effects on the inhibition of invasion and characterize pathways involved in transendothelial migration.
Figure 4. Treatment effects on leukocyte transendothelial migration. Inhibition of CXCL12 (SDF-1α) driven T cell migration by AMD3100 (A), BIRT377 (C) and neutralizing ICAM-1 (D). Analysis of AMD3100 pharmacological response (B). Migration across the endothelial layer was at least partially dependent on ICAM-1, as neutralizing ICAM-1 antibodies, and treatments with allosteric inhibitor of LFA-1, BIRT 377, significantly inhibited SDF-1α mediated diapedesis of T cells.
Figure 5. Profile inhibitors of tumor cell invasion. Incucyte® high-definition images of Incucyte® Nuclight HT-1080 fibrosarcoma cells migrating across a 2D substrate and invading through a 3D basement membrane extract (BME) biomatrix in response to a serum gradient – note the differential morphology and invasive filopodia-like projections that extend into the ECM. Time-course plots reveal that cell invasion through the BME matrix is slower than migration and that invasion, but not migration, is inhibited by the matrix metalloproteinase inhibitor, GM6001 in a concentration dependent manner.
Chemotaxis Assay Technical Resources
Reagents-Consumables-Brochure-en-L-8000-0738-B00-Sartorius.pdfPDF | 6.1 MB
Incucyte® Chemotaxis Cell Migration Assay ProtocolPDF | 772.4 KB
Detailed Incucyte® Chemotaxis Cell Invasion Assay ProtocolPDF | 596.2 KB
Incucyte® Chemotaxis Transendothelial Migration Assay ProtocolPDF | 633.5 KB
Differentiated THP-1 Incucyte® Chemotaxis Migration ProtocolPDF | 98.2 KB
Detailed T Cell Incucyte® Transendothelial Migration Chemotaxis ProtocolPDF | 99.8 KB
Incucyte® Chemotaxis Assay Protocol for Macrophages Protocol ManualPDF | 108.2 KB
Detailed Incucyte® Chemotaxis Assay Protocol for Jurkat Cells ManualPDF | 108.0 KB
Chemotaxis Frequently Asked Questions
The Incucyte® Scratch Wound Assay measures the movement of cells into a cell-free zone in the absence of a chemotactic gradient. You can use the Incucyte® Woundmaker Tool to create a precise wound in each well and monitor wound closure as cells move either across a substrate (migration) or through a 3D gel matrix (invasion).
The Incucyte® Chemotaxis Assay measures directed cell migration in response to a chemotactic gradient. You can use the Incucyte® Live-Cell Analysis System to investigate treatment effects on chemotactic profile. The system is suitable for adherent and non-adherent cells.
No. The optical quality of the membrane surfaces in existing Boyden chambers is not amenable to Incucyte® imaging or image processing algorithms.
No. The Incucyte® Clearview cell migration insert has been specifically designed to work with the Incucyte® Clearview reservoir plate.
Version 2015A Rev 1 or later. The Incucyte® Chemotaxis Analysis Software Module is not compatible with prior software versions. If you wish to receive a software upgrade, please reach out to your point-of-sale contact.
Can I measure migration of non-adherent cell types using the Incucyte® Clearview Cell Migration Plate?
Yes. Incucyte® Clearview Cell Migration Plates have been extensively validated with non-adherent cell types. To ensure uniform cell settling within each well after seeding, we recommend allowing the plate to sit at room temperature for 45-60 minutes after plating. The Chemotaxis (Top Only) “Analysis Job Type” is recommended for quantifying non-adherent cell migration.
Key variables to optimize include membrane coatings, assay medium and cell density. Recommended experimental conditions for a range of cell types - including cancer cells, vascular cells and leukocytes - can be found in the assay protocol.
Yes. Top and bottom membrane surfaces can be coated independently or together. Recommended volumes are 20 μL for the top side of the membrane and 150 μL for the bottom side.
Yes. We recommend using a nuclear restricted fluorophore (as opposed to cytoplasmic) to ensure neighboring cells can be differentiated and counted as individual objects following image analysis. Fluorophore excitation and emission spectra must be compatible with Incucyte® fluorescent channels.
Yes. By labelling different cell types with fluorescent probes and using the phase, green, red, orange or near-IR red data acquisition channels of Incucyte® Live-Cell Analysis System it is possible to quantify chemotaxis of more than one cell type in co-culture.
Yes. The Incucyte® Live-Cell Analysis System does not disturb your cells during the assay. Once the experiment is completed, simply remove the medium from the wells to perform additional analyses.
Yes. Multiplexed measurements of cell health can be made using any unused imaging channel. The Incucyte® Live-Cell Analysis System supports HD phase-contrast and green, red, orange and near-IR fluorescence automated imaging modes.