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DIY breakdown: Advantages and Disadvantages of Do-it-yourself Flow Systems

Mechanical forces have been shown to impact cell phenotype and function in vivo. The most prolific of these forces is shear stress, induced by fluid movement throughout the body. Shear stress has been demonstrated to impact polarization, alignment, differentiation, and function of various cell types, including endothelial cells, leukocytes, and stem cells.

Despite the impact of shear flow, the majority of in vitro cell culture experiments are still conducted in static systems, such as plates and flasks. This creates a challenge when attempting to translate the results of in vitro experiments to an in vivo environment, as cells may not look or act the same. To overcome this gulf, some scientists have begun to use do-it-yourself flow cells. Although these DIY flow cells improve the physiological relevance of in vitro experiments by attempting to mimic an in vivo environment, there are several drawbacks that are outlined below.

endothelial cells with and without flow

Figure 1. Representative images showing endothelial cells cultured without (A) and with shear flow (B).

DIY Drawbacks

  1. Piecemeal system

DIY flow cells are exactly that, do-it-yourself. That means multiple components, including pumps, tubing, slides, and more must be assembled according to the experimental needs. This could mean hours of checking and double-checking parts , ordering parts from multiple companies, or, as is the case with a bubble trap, building parts yourself.        

 

  1. Carryover and contamination

With most DIY flow systems, the fluid that runs across the cells is carried from a bulk container to and from the flow chamber by a tube or set of tubes. Although effective, this means that tubing must be thoroughly cleaned or changed between experiments to avoid carryover or contamination from previous experiments. This creates a burden, as investigators must either spend money on new tubing or spend valuable time cleaning tubing and waiting for it to dry.

 

  1. Throughput limitations

Collecting data quickly is a key component in meeting deadlines for publications, grant applications, and more. Although some slide-based flow chambers offer > 1 well throughput, the per slide throughput is typically limited to 6 chambers or less. This means that to achieve “high throughput” experimentation, a complicated setup is required (Figure 2). This type of setup is time-consuming, requires a large footprint in the lab, and is expensive.

DIY shear flow system

Figure 2. Representative “DIY” flow cell setup. A typical setup includes flow chamber(s), reservoirs, tubing, 3 way stopcocks, male and female luer adapters, a pressure controller, and a microscope with a camera for imaging.


Affordable Alternative to DIY Flow System

With these issues in mind, a viable alternative to slide-based shear flow systems is extremely desirable. The BioFlux family of shear flow systems separate themselves from slide-based shear flow systems by using microfluidic channels embedded into the bottom of 6, 24, or 48 well glass bottom plates. The microfluidic design combined with leveraging pneumatic pressure to precisely control shear flow has several key advantages over traditional slide-based shear flow systems.


BioFlux Advantages

  1. A Complete System

BioFlux is a complete, integrated solution, including hardware, software, and consumables. Unlike DIY flow cells, everything is included and engineered to work together seamlessly.

  1. Carryover and Contamination free

BioFlux pioneered the concept of contactless pneumatic pumping; samples and solutions only touch the single-use consumable, so starting the next experiment is as easy as filling a new plate. No need to change or sterilize pumps or tubing between experiments.

  1. High Throughput

BioFlux plates are available in Society for Biomolecular Screening (SBS) standard-sized 6, 24, or 48 well plates. This means that any lab equipment that works with standard plates can also be used on BioFlux plates, including multichannel pipettes. A standard 48-well BioFlux plate can run 24 simultaneous experiments. In addition, BioFlux offers a Quattro addon that enables up to 4 plates to run simultaneously, speeding drug discovery and other investigations that require high throughput assays.

  1. Experimental Flexibility

A major advantage of the BioFlux 200 and BioFlux 1000Z systems is the ability to create two independent parallel flow streams when using a 24-well plate. This enables the generation of a gradient between the two fluids. This gradient allows for complex experimentation such as cell migration/invasion, chemotaxis, and wound healing assays that are either not feasible or not possible with conventional flow cells.

Advantages of DIY Flow Systems

  1. Price

Although the BioFlux family of shear flow systems provides key advantages over DIY flow cells, typically, a DIY system can be assembled for a lower price than BioFlux 200 or BioFlux 1000Z. However, the recent introduction of the BioFlux One system has eliminated this price gap. Although not as experimentally flexible as BioFlux 200 and BioFlux 1000Z, BioFlux One is a complete system with all the functionality of a piecemeal DIY system but with higher throughput and a cleaner environment.

  1. Cell Recovery

Another advantage of DIY flow cells in comparison to BioFlux systems is the recovery of adherent cells. Some DIY systems can be disassembled for recovery and further analysis of adherent cells. Standard glass bottom BioFlux plates require that adherent cells be released by flushing the microfluidics with an enzyme, such as trypsin. While this type of release can enable unique assays, such as wound healing, it may negatively impact downstream analysis. However, a BioFlux custom substrate plate allows for the attachment of “coupons,” made of a material of the user’s choice, to the bottom of the plate. These coupons can be removed after the experiment for recovery of adherent cells.

BioFlux system with microscope CMS logo

A BioFlux 200 shear flow system with an inverted microscope and high-speed camera.

Flow chamber assay capability comparison 

BioFlux vs DIY assay comparison

Conclusion

Adding shear flow to experiments greatly enhances the physiological relevance of cell-based assays; however, investigators should carefully consider the current and future needs of their experiments before purchasing or assembling a shear flow system. Taken together, these facts highlight the key advantages and disadvantages of both BioFlux shear flow systems and DIY flow cells.

Dr. Anson Blanks completed his BS in exercise physiology at East Carolina University in 2003. Dr. Blanks then attended Appalachian State University, where he earned his Master of Science in clinical exercise science in 2009. After working as a clinical exercise physiologist in a cardiopulmonary rehabilitation center in Washington, DC, Dr. Blanks decided to pursue a career in scientific research. He attended Virginia Commonwealth University, where he completed his Ph.D. in rehabilitation and Movement Science in 2018. After spending several years as a research and development scientist in biotechnology industry, Dr. Blanks is now a scientific marketing manager for Cell Microsystems in Durham, NC.

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