Forces such as tension, compression and shear stress are physiologically important regulators of cell responses in vivo. Differences in blood vessel diameter and geometry across the arterial tree account for a wide physiological range of flow rates, flow profiles and shear stress in the circulatory system. Interstitial flow through porous tissue networks creates pressure and shear stress gradients. Fluid flow over cells cultured in microfluidic perfusion chambers provides key mechanical stimuli that are largely absent from traditional culture methods. Importantly, parameters such as flow rate and shear stress can be carefully controlled.
In this application note we describe how to recirculate medium for dynamic cell culture, use the OB1 pressure controller to maintain a precise flow rate of medium over cells in the perfusion chamber and hook up a MUX-Injection valve to enable recirculation of medium between two reservoirs unidirectionally over your cells. Recirculation enables higher flow rates to be maintained for multiple days, medium conditioning and studies of shear stress. Read also our application notes about automated cell seeding, microfluidic perfusion and cell staining for dynamic cell culture.
Some applications of dynamic flow in cell culture include:
Cell and biology pack
Microfluidic chamber designed for perfusion (IBIDI µSlide I Luer)
Flow controller OB1 Mk3+
Mux-Injection
Microfluidic bubble trap
Flow sensor
Tubing, fittings and reservoirs
Alternate between MUX-Injection valve POS 1 and POS 2. Liquid flows from one reservoir (pressurised) to the other (not pressurised). Flow is unidirectional through the chip.
Seed cells manually using a pipette into a microfluidic chip. Leave 6-24 h to attach.
Set valve: Set MUX-Injection valve to position 1.
Flow: Start flow from reservoir 1 (pressurise using OB1 channel 1). Reservoir 2 will collect the flow-through from the chip. Perfuse for desired time.
TIP: Both reservoirs will need to contain medium at the beginning: fill reservoir 1 (initial liquid source) with about 70% of the total volume and reservoir 2 (initial liquid collector) with 30%. Leave some volume in reserve in reservoir 1 before switching liquid sources to avoid introducing air into your perfusion chamber.
Switch valve: Switch MUX-Injection valve to position 2.
Flow: Start flow in reservoir 2 (pressurise using OB1 channel 2, depressurise reservoir 1). Reservoir 1 will collect the flow-through from the chip. Perfuse for desired time.
Repeat: Continue cycling between MUX-Injection valve positions to perfuse cells in culture for desired overall time.
MCF7 cells grown with continuous medium recirculation (10 µL/min, 24 h). Stained with Hoechst/ rhodamine phalloidin (fixed with PFA, 4%, permeabilized with BSA, 1%/ Triton X, 0.1%).
Application note written by Lisa MUIZNIEKS, Subia BANO, Camila BETTERELLI GIULIANO and Jessica AYACHE.
This work was done with the support from the French Agence Nationale de la recherche (ANR) in the frame of ERA-NET JPco-fuND 2019 (Orgtherapy).
Do you want tips on how to best set up your microfluidic experiment? Do you need inspiration or a different angle to take on your specific problem? Well, we probably have an application note just for you, feel free to check them out!
Biofilm testing using a simple microfluidic chip channel for in situ observation of their development under flow conditions.
Microfluidics for microscopy imaging in plant biology allows to observe, in vivo, the biological response of plant roots to various stimuli.
This application note describes how co-culture of different cell types in separate but interconnected chambers is possible in a microfluidic platform
This application note explains how to study bacteria adaptation to stress and environmental changes such as antibiotics.
In this application note we describe how to set up medium recirculation by using microfluidic valves
In this application note we describe how to stain cells for dynamic cell culture with different microfluidic setups.
In this application note we describe how to do cell perfusion for dynamic cell culture and a way to enable uni-directional recirculation of medium.
A simple guide to do dynamic cell culture by automating cell seeding in a microlfuidic chip
This application note proposes a microfluidic cardiac cell culture model (μCCCM) to recreate mechanical loading conditions observed in the native heart (in both normal and pathological conditions) by using an Elveflow OB1 pressure and flow controller.
In this application note, we will describe how to perform an automated and fast medium switch thanks to the Perfusion Pack.
Medium switch is widely used in cell biology. One application is the study of cell behavior under given flow conditions for different samples. In this tutorial, we walk you through the steps of a fast and stable medium switch using IBIDI© flow cells.
Fluorescence reader for microfluidic qPCR: faster, more sensitive and less expensive than most optical microscopes, it is a smart alternative for real-time fluorescence measurements of your on-chip qPCR signal.
Prostate cancer is the second leading cause of cancer-related death for men. Circulating tumor cells (CTCs) are considered as a marker of early cancer diagnosis and disease severity. Their screening in blood is thus crucial to detect metastatic stage in cancer patients.
Until recently, microfluidic devices have been employed to support tissue-engineering experiments on basal lamina, vascular tissue, liver, bone, cartilage and neurons as well as organ-on-chips.
This application note presents how to perform cell culture on chip using the Cell and Biology Pack and the MicroSlides developed by ALine Inc.
Get a quote
Name*
Email*
Message
Newsletter subscription
We will answer within 24 hours
By filling in your info you accept that we use your data.
Collaborations
Need customer support?
Serial Number of your product
Support Type AdviceHardware SupportSoftware Support
Subject*
I hereby agree that Elveflow uses my personal data Newsletter subscription
How can we help you?
Message I hereby agree that Elveflow uses my personal data Newsletter subscription