Millipore Sigma Vibrant Logo
 
 

Applications of the CellASIC® ONIX Microfluidic Platform

Request Information

Discover our latest application notes featuring the CellASIC® ONIX2 Microfluidic Platform




Merck:/Freestyle/BI-Bioscience/Cell-Culture/cellASIC/cellasic-application-dapi-hif1a.jpg
Chemoresistance changes parallel HIF1a protein levels. Data is presented as fluorescence intensity on a per cell basis (normalized). Total cells (DAPI); hypoxia-responsive fraction (inset images, 20X).

NEW! An integrated platform for real-time dynamic culturing and analysis of hypoxia with single cell resolution
Hypoxia is a common characteristic of solid tumors and is linked to chemo-resistance and poor clinical outcomes. Performing detailed studies of the hypoxic response in cancer requires precise control of the cell culture environment, combined with real-time imaging. In this study, we assess the hypoxic response of diverse cancer cell lines using the CellASIC® ONIX2 Microfluidic System, which captures live-cell images and video under precisely controlled culture conditions. We observed that hypoxia affected response to cytotoxic agents, mechanisms of cell death, and invasive capacity. Similar studies will help broaden our mechanistic understanding of hypoxia and lead to improved interventions in cancer.

Download Application Note




Merck:/Freestyle/BI-Bioscience/Cell-Culture/cellASIC/FOX_Graph_1-500px.jpg
Changes of the fluorescence intensity in the tracked nuclei upon treatments.

Establishing a Time Course for Translocation of FOXO4 in Live Cells Using a Novel Microfluidic Culture Platform Assay
This study establishes the first time-course analysis for translocation oof FOXO proteins following deactivation of the PI3K-Akt pathway, by combining live cell analysis, fluorescently-tagged reporter cells, and the unique microenvironmental control capabilities of the CellASIC® ONIX2 system. The assay method described here provides quantitative information on both FOXO4 nuclear import and export, and thus provides a platform for the discovery of new targets and therapeutic compounds in cancer as well as other diseases.

Download Application Note




Primary neuron culture and analysis facilitated by the CellASIC® ONIX2 Microfluidic Platform
Immunocytochemistry of the rat primary cortical neurons cultured, stained and analyzed on the microfluidic platform.

Primary Neuron Culture and Analysis Facilitated by the CellASIC® ONIX2 Microfluidic Platform
Perturbation analysis of live neurons is crucial for fully understanding the nervous system and requires spatiotemporal microenvironment control of cultured neurons. Advances in microfluidics have enabled such studies, but most microfluidic analyses of neurons require a prohibitive commitment of resources. The CellASIC® ONIX2 Platform is a novel, optimized microfluidic platform for long-term culture of primary neurons to monitor dynamic cellular processes in real time, enabling users to automate changes to culture conditions while offereing a superior platform for visualization and no specialized expertise required.

Download Application Note




In-plate immunocytochemistry using anti-cytokeratin 19, an intracellular protein, in fixed and permeabilized PC-3 cells. Scale bar = 100 µm..
In-plate immunocytochemistry using anti-cytokeratin 19, an intracellular protein, in fixed and permeabilized PC-3 cells. Scale bar = 100 µm.

Long-term Cell Culture and In-Plate Staining:
Demonstration of long-term culture and in-plate staining protocols using the CellASIC® ONIX2 Microfluidic Platform

The ability to analyze cell responses to perturbations over time greatly extends the precision and biological relevance of in vitro studies. However, there are technical challenges to successful long-term cell culture and analysis. Not only is it difficult to control the temperature and gas composition of the cell environment during the course of the experiment without impeding optical access to the cells, but it is also challenging to maintain sufficient nutrient supply and outflow of waste to keep cells healthy. The easy-to-use CellASIC® ONIX2 Microfluidic Platform delivers precise control of perfusion, temperature and gas conditions, facilitating long-term cell culture and enabling automated, in-plate cytochemical analysis using minimal reagent volumes. Here we demonstrate the long-term culture of multiple cell lines using automated perfusion protocols set up with intuitive CellASIC® ONIX2 FG Software, comparing the results to those obtained using traditional chambered slides.

Download Application Note
 



Views of (A) an invasive strain and (B) a non-invasive strain of E. coli after exposure to human HT-29 cells, washout, and perfusion culture. Bacteria expressed mCherry, and HT-29 cells stained with Calcein AM. Panel (A) was acquired with a 100X objective lens, and panel (B) was acquired with a 60X objective lens.
Views of (A) an invasive strain and (B) a non-invasive strain of E. coli after exposure to human HT-29 cells, washout, and perfusion culture. Bacteria expressed mCherry, and HT-29 cells stained with Calcein AM. Panel (A) was acquired with a 100X objective lens, and panel (B) was acquired with a 60X objective lens.

Host Pathogen:
Long term, live cell analysis of host-pathogen interactions using the CellASIC® ONIX2 System

The proper study of host-pathogen interactions requires careful control of the cell environment to simulate physiologic conditions. An in vitro model that can replicate infection parameters, including flow rate, exposure time, solution type, and timed introduction of therapeutic agents while sustaining long-term live cell analysis requires experiments that are difficult or impossible with standard methods. The CellASIC® ONIX2 Microfluidic System is well-suited for host-pathogen studies by providing a stable, long-term culture environment for host cells with controlled pathogen exposure. Here we demonstrated a host-pathogen experiment using human intestinal cells infected with engineered E. coli strains.

Download Application Note 



Comparison of per-cell EGFR expression of MCF-10A cells cultured in the CellASIC® ONIX2 System (M04S plate) vs. in standard Petri dish.
Comparison of per-cell EGFR expression of MCF-10A cells cultured in the CellASIC® ONIX2 System (M04S plate) vs. in standard Petri dish.

Long-Term Cell Culture and Gene Expression Analysis:
Microfluidic perfusion enables long-term cell culture, precise microenvironment control and gene expression analysis

The analysis of living cells in vitro is critical to understanding basic biology signaling pathways, drug effects, and disease models. Yet, technology for environment control of living cells during analysis has not advanced significantly since the Petri dish. A growing body of evidence indicates that the cellular environment, or "niche", is just as important as genetic factors for determining cell phenotype. Therefore, a method for providing more accurate, dynamic control of living cells has the potential to dramatically advance the state-of-the-art for live cell analysis. The CellASIC® ONIX2 Microfluidic System, in conjunction with the CellASIC® ONIX2 Microfluidic Plate provides perfusion-based microenvironment control for long-term, high quality, live cell analysis.

Download Application Note 



Dynamic Autophagy Assay
Schematic of live cell analysis for autophagy of LC3-GFP-expressing CHO cells. First, medium was perfused to establish fluorescence baseline (left). Serum starvation and the lysosome inhibitor CQ1 were then introduced to trigger autophagosome accumulation (center). When cells were returned to standard growth medium, autophagosomes underwent lysosomal degradation (right).

Dynamic Autophagy Assay:
A dynamic live cell assay platform to elucidate the mechanisms underlying autophagy

Autophagy is a complex cellular process essential for cell survival under stressed conditions. However, the dynamics of autophagy during a cell's stress and recovery phases are not fully understood. Here, we report a dynamic, live cell assay to monitor both the rate of autophagasome formation and changes in lysosomal degradative processes during autophagy. The CellASIC® ONIX2 Microfluidic Platform combined live cell analysis with cell lines stably expressing fluorescently-tagged markers specific for autophagosomes to enable the cellular dynamics of autophagy to be visualized at a single cell level in real time.

We used the CellASIC® ONIX2 Microfluidic Platform with M04S plates to develop a dynamic cell-based assay for monitoring autophagy and elucidating the dynamics of lysosomal degradation. LC3-GFP CHO reporter cells were subjected to nutrient starvation or hypoxic stresses for a designated time period followed by reintroduction of normal growth conditions. The time course of autophagy was visualized under a fluorescent microscope.

Download Application Note 




The CellASIC ONIX2 Microincubator utilizes an innovative recirculating heat exchanger manifold to regulate temperature and gas composition of the microfluidic chambers.
The CellASIC® ONIX2 Microincubator utilizes an innovative recirculating heat exchanger manifold to regulate temperature and gas composition of the microfluidic chambers.

Live Cell Analysis and Hypoxic Culture:
Microincubator for Long-Term, Live Cell Analysis and Hypoxic Culture

Live cell analysis technology has made extraordinary progress in recent years, with improvements in microscopy, cellular probes, and genetic engineering. The ability to perform live cell experiments within microfluidic chambers further extends the precision and biological relevance of in vitro studies. One of the major technical challenges for long-term cell analysis is controlling the temperature and gas composition of the cell environment during the course of the experiment without impeding optical access to the cells. The cost and inconvenience of performing experiments in static, hypoxic conditions have, until now, hindered the study of cell behavior in low-oxygen environments. The CellASIC® ONIX2 Microfludic platform is a promising basis on which to design an improved live cell incubation system with dynamic control.

Download Application Note 



High density seeding of NSCs in a microfluidic chamber under conditions of mild hypoxia causes formation of neurospheres. Shown is a neurosphere formed after 72 hours, stained for Sox2 and nestin expression following 'on-chip' immunostaining protocols.
High density seeding of NSCs in a microfluidic chamber under conditions of mild hypoxia causes formation of neurospheres. Shown is a neurosphere formed after 72 hours, stained for Sox2 and nestin expression following "on-chip" immunostaining protocols.

Rat Neural Stem Cells (NSCs):
CellASIC® ONIX2 live cell analysis platform for neural stem cell microenvironment control

Neural stem cells (NSCs) are sensitive to microenvironmental cues, including cell-cell contact, cell-ECM interaction, nutrient and waste transport, as well as environmental oxygen composition. How these parameters affect the stem cells' morphology, proliferation, and differentiation remains an open area for research. In this study, we demonstrated how the CellASIC® ONIX2 Microfluidic Platform with its microfluidic cell culture capabilities provides multi-parametric microenvironment control for NSC studies.

Download Application Note 



After 24 hours, 3D cell clusters were imaged in sharp focus using the CellASIC ONIX2 Microfluidic Platform. Scale bar = 100 µm.
3D cell cultures in sharp focus using the CellASIC® ONIX2 Microfluidic Platform. Scale bar = 100 µm.

3D Cell Culture:
Three-dimensional culture and assessment of drug-induced cell death using the CellASIC® ONIX2 Microfluidic Platform.

Cultured cells have proved to be convenient, powerful model systems for elucidating biological processes and for testing the safety, efficacy and mechanism of therapeutic candidates. Current strategies for 3D cell culture include growing cells in hanging drops, in a natural or synthetic 3D matrix on biodegradable polymers in a cross-linked hydrogel or in porous synthetic scaffolds. Even in these advanced platforms, if subjected to static conditions of gas, nutrient medium and waste buildup, they are limited by the inefficient mass transport between the inside and outside of the 3D cell structures. To overcome the challenges of mass transport in 3D culture, the use of microfluidics has gained in popularity. The CellASIC® ONIX2 Microfluidic System enables perfusion-based microenvironment control in the study of drug-induced cell death of 3D cultures of MCF7 cells in Matrigel®.

Download Application Note 



Tracking migration of individual MDA-MB0231 cells in response to a stable FBS gradient enables quantitation of individual cell speed and directionality.
Tracking migration of individual MDA-MB0231 cells in response to a stable FBS gradient enables quantitation of individual cell speed and directionality.

Cell Migration Analysis:
Automated live cell analysis of cell migration across a microfluidic-controlled chemoattractant gradient

Cell migration is stimulated and directed by interaction of cells with the extracellular matrix (ECM), neighboring cells, or chemoattractants. During embryogenesis, cell migration participates in nearly all morphogenic processes ranging from gastrulation to neural development. In the adult organism, cell migration contributes to physiological and pathological conditions, and is central to development of therapeutics affecting wound healing and tumor metastasis. Mechanisms of migration can be understood by analyzing cellular response to modulators of migration, therefore, techniques for the precise quantification of cell migration behavior in response to a gradient have become central to life science research. The most widely accepted cell migration assay is the Boyden chamber assay using chemoattractants. The CellASIC® ONIX2 Microfluidic Platform offers a microfluidic gradient plate to enable precision-controlled chemoattractant diffusion across perfusion barriers, enabling a quantitatively defined diffusion gradient, stable enough for long-term, live cell analysis over the course of days. Using the power of long-term, live analysis of cells, we studied the effect of a serum gradient on metastatic breast cancer cell migration distance, velocity and degree of chemotaxis.

Download Application Note 



Other Applications of CellASIC® Technology:

  • Cell response over time
  • Chemotaxis/migration
  • Drug dose/response
  • Hypoxic Conditions to Mimic Tumor Microenvironments
  • Bacteria Single Cell Analysis
  • Yeast Single Cell Analysis