In-vivo Conditions for Live Cell Assays
Many physiological processes take place under flow conditions: Blood flowing through the vasculature; cancer cells circulating throughout the body; plaque forming on teeth under the presence of saliva flow. It is now well established that physiological flow has a profound impact on many biological studies, yet a lot of research is still studied in vitro without the presence of flow. BioFlux Systems give you the ability to introduce flow to your research and drug discovery experiments effectively emulating in vivo conditions and getting closer to understanding what’s happening under more physiological conditions.
Offering the physiological relevance of laminar flow chambers with the throughput and convenience of standard well plates, BioFlux systems have many features and benefits including:
- High Speed: With automated experimental control, multiplexing up to 96 simultaneous experiments, and sophisticated data analysis software simplifying and accelerating complex functional assays, large projects can be effectively reduced from months to days.
- High Quality: Glass bottom plates with superior image quality from microscopes and high content imagers provide unparalleled image quality.
- Simple to Use: Well plate microfluidic design improve reliability and reproducibility while enabling precise control of critical parameters.
- Modular Design: A BioFlux system can be added to any existing inverted microscope setup instantly expanding its capabiliti
BioFlux systems come in different configurations and have been featured in multitudes of publications, presentations and webinars.
Bacteria and fungi organized into biofilms are highly refractory to known antibiotics and biocides. In order to develop new solutions to combat biofilms, screening technologies must be designed to grow biofilms under conditions that represent an in vivo or in situ community while maintaining the ability to add and observe the effects of compounds. BioFlux offers a comprehensive solution for running both fundamental biology assays as well as anti-microbial compound screens.
- Biofilm growth
- Mutant screens,
- Antibacterial or Antifungal screening
- Host-pathogen interactions
- Adhesion strength
(from left to right)
- An example of Pseudomonas aeruginosa growing over 24 hours.
- Staphylococcus epidermidis growing a biofilm over 16 hours in a BioFlux plate
- PlasDIC imaging used to visualize yeast adhering to a HUVEC monolayer. Notice the start of hyphae projections
- Co-culture of Candida albicans expressing GFP and mCherry: Grown at 0.5 dyn for 17 hours. Imaged every 5 minutes and played at 3 frames per second.
Proper immune function depends on the interconnected relationships of many diverse cell types. When improperly functioning, these cell-cell interactions can lead to allergy, asthma, autoimmune and infectious disease. Immune surveillance in large part occurs within the vasculature in the presence of constant flow, where cell-cell and cell-ligand interactions are influenced by in situ forces. BioFlux provides the ability to control the timing of these interactions as well as the shear rate.
- Transmigration migration
- Cell adhesion and rolling
- Wound healing
(from left to right)
- Lymphocytes (in white) migrating through a monolayer of endothelial cells (darker cells) over 8 hours
Chemotaxis assay established using the 24 well dual plate where human neutrophils migrated to fMLP perfused through inlet A (top) while control media was perfused in inlet B (bottom). Imaged in FITC channel (cells were stained with CellTracker).
Jurkat T cells rolling over E-selectin at 2 dyn
Latex beads demonstrating how pulsatile flow is a physiologically relevant method of vascular perfusion.
Platelet Function: Platelet aggregation, or thrombosis, occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. It can be triggered in the presence of shear flow and is dependent on many biochemical interactions present in whole blood. The BioFlux system enables simulation of vasculature shear rates up to 200 dyne/cm2 (5000 s-1) using whole blood, platelet-rich plasma or other cells of interest. Experiments can be run on purified matrix proteins (von Willebrand factor, collagen, etc.) or endothelial cell monolayers. The system uses as little as 20μL of reagents per experiment.
Vascular Biology: The endothelium is a dynamic mediator of many physiological phenomena and the crossroads for immune system function, cancer metastasis and atherosclerosis among others. Endothelial cells have been shown to alter their morphology and gene expression in the presence of the shear flow they routinely see in the vasculature. As such, the relevance of endothelial cell culture and downstream assays increases significantly with the presence of flow.
- Platelet aggregation and adhesion Thrombosis
- Migration and invasion
- Atherosclerosis models
(from left to right)
- CalceinAM Labeled Platelet Aggregation on Collagen 1:Whole blood labeled with calcein AM flowing over collagen 1 at 40 dyn over 5 minutes.
- Blood-Endothelial Cell Interaction Using RICM: Interference reflection microscopy used to image the interaction when whole blood flows over endothelial cells.
- Dual Flow Platelet Aggregation: Whole blood flowing in our dual flow 24 well plates where inlet A (top) had control blood and inlet B (bottom) had blood treated with anti-gpIIb
- Rolling Platelets: In whole blood where platelets have been stained with calcein AM, platelets roll over channels coated with VWF.
Understanding the molecular and cellular mechanisms of cancer progression, from primary malignancy to metastatic disease, is the key to development of successful treatments. Cancer cells travel through the vasculature under flow and interact with cells in the endothelium. BioFlux provides a physiologically-relevant in vitro model for studying these interactions under controlled shear flow conditions.
(from left to right)
- Cancer Cells Invade Matrigel : The 24 well dual plate was used to measure the invasive properties of HT 1080 fibroscarcoma cell into matrigel. Matrigel was perfused through inlet B (bottom) while PBS was perfused in inlet A (top) to establish an invasion assay. Cells invaded over 12 hours.
- PC3 Cells Invade Lymphatic Endothelial Cells in 3D: The Nikon Imaging Center (UCSF) captured a 3D stack image of human prostate PC-3 cells (green) invading through a monolayer of endothelial cells (red).
Stem cell research has the potential to produce novel treatments for previously incurable diseases and injuries. The application of controlled shear flow to undifferentiated embryonic stem cells promotes enhanced expansion of cell lines. Shear stress can be used as a stimulus for differentiation especially for cell types that naturally respond to physiological shear such as endothelial cells. Differentiation of cells into specific cell types and subsequent production of biomaterials is also facilitated by mechanical forces such as shear.