Optimal Cell Disruption
Utilizing intracellular contents (proteins, organelles, DNA/RNA, enzymes and Adeno-Associated Virus (AAVs) Vectors) found and/or grown inside cells is the next generation of drug development. For cells that do not secrete these intracellular contents, it is vital to lyse the cell to liberate the contents. It is important to not denature intracellular components by unnecessary elevation of temperature or excessive shear rates. Microfluidizer® processors are extremely efficient at rupturing all cell types with minimal denaturation of the delicate intracellular contents. Our processors are great for the homogenization of cells because they are tough on cells and gentle on intracellular contents. Microfluidizers® are ideally suited for effectively rupturing cells requiring different levels of shear — including bacterial, mammalian, plant, insect, fungi, algae and yeast cells — while ensuring high protein recovery. These capabilities allow researchers to use the lowest shear rates possible to reach target rupture rates while avoiding denaturation of intracellular contents.
Proven Processing Results
Microfluidizer® processors provide many demonstrable advantages over all other cell disruption methods and equipment — for both lab and production volumes.
|E. Coli Cell Rupture|
Bacteria Lysis (E. Coli) Before and After processing on the Microfluidizer® for 1 pass achieving >95% lysis
|Yeast Lysis (S. Pombe)|
Yeast Lysis (S. Pombe) Before and After processing on the Microfluidizer® for 5 passes achieving ~95% lysis
Microfluidizer® Benefits for Cell Disruption
Highest Protein Integrity Recovery
Precisely controlled shear rates enable Microfluidizer® customers to use the minimum pressure required to rupture the target level of cells, while keeping proteins intact. Compared with other cell disruption techniques, the Microfluidizer® yields several times the amount of recoverable, usable protein. See a compilation of peer reviewed publications that highlight the performance and advantages of using a Microfluidizer for cell disruption
Cooling is extremely important in cell disruption because cell contents are typically temperature sensitive. Immediately after rupturing, the Microfluidizer® heat exchanger minimize the amount of time the sample experiences elevated temperatures and the lower temperatures combined with shorter processing times result in reduced denaturing and increased yields.
Ease of Use
Microfluidizers® were designed with convenient homogenization of cells and productivity in mind — that’s why they are simple to operate and easy to clean. Using a Microfluidizer in a lab requires no specialized skills or knowledge and initial training is provided during machine install. Customers appreciate how little maintenance is required, especially when compared to high-pressure valve homogenizers that have valves that need to be disassembled and cleaned manually, which is important especially in a multi-user environment to limit cross contamination.
Simple Downstream Processes
The Microfluidizer® breaks cells gently yet efficiently, resulting in large cell wall fragments. The large fragments are easier to separate from the much smaller cell contents. Filtration times are shorter and the need for centrifugation is reduced.
Microfluidizer pressure and the number of passes can be set to shear DNA if desired. Shearing the DNA makes downstream pipetting of small volumes of cell lysate simpler and more accurate.
Processes at a Constant, Controlled Shear Rate
Continuous processing at constant pressure and shear rates ensures that all cells receive the same amount of energy input. With sonication, cells closer to the probe receive exponentially more energy than cells farther from the probe; batch-processing methods provide little control of energy uniformity to each cell. Some cells remain unbroken, others die or the intracellular contents denature due to the uncontrolled temperature rise of sonicator probes. Monitoring the temperature of a volume of cells cannot tell you what has happened to the majority of cells in a sample. This is why Microfluidizers give higher yields than sonicators. Processing at lower shear rates make it possible to rupture mammalian cells with high efficiency for the harvest of Adeno-Associated Virus (AAVs) Vectors for gene therapy.
The image above outlines the shear rate required to rupture various cell types. The Microfluidizer can be precisely tuned by changing the pressure and Interaction Chamber (IXC) configuration to match and not exceed the necessary shear rate for the cells being processed.
Microfluidizer® machines offer media-free, negligible-wear processing that eliminates contamination of your sample.
Unlike other technologies used to rupture cells, Microfluidics guarantees scale-up from lab and pilot volumes to full-scale production. The ability to scale up from lab to production volumes is a highly valuable tool for cell homogenization researchers — and an area where our processors shine. Unlike Microfluidizers®, high-pressure valve homogenizers involve changes in the way the cells are ruptured to accommodate the higher flow rates, which results in inconsistency when scaling up. When you use a Microfluidizer®, scale-up performance is guaranteed.
Microfluidizers® are capable of handling a wide range of cell types by optimizing pressure and cooling.
Small Sample Volumes
Our LV1 Low Volume Lab Machine is capable of cell homogenization in samples as small as 1 ml.
Advantages in the Lab
Lab Scale Microfluidizers® process cells rapidly (up to several hundred ml/min) from small sample volumes (as small as 14 ml to several liters). Our machines utilize advanced technology to rupture even the most challenging cells, and enable multiple research groups to use the processor for diverse applications.
Advantages During Production
The ability to scale up from lab to production volumes is a highly valuable tool for cell homogenization researchers — and an area where our processors shine. Unlike Microfluidizers®, high-pressure homogenizers involve changes in the way the cells are ruptured to accommodate the higher flow rates, which results in inconsistency when scaling up. When you use a Microfluidizer®, scaleup performance is guaranteed.
Cell Disruption Methods
Advantages of Microfluidizers® Compared to Other Cell Lysis Techniques
High Pressure Valve Homogenizers: After a Microfluidizer®, a high-pressure valve homogenizer for cell disruption is the next best alternative. Prices are typically comparable, although cooling, cleaning, valve wear and scalability can be issues. The Microfluidizer® provides superior quality and usability of ruptured cell suspensions for increased yield.
French Press: When using a French Press for cell disruption, a manually controlled valve releases the pressurized fluid from a pressure cell, resulting in cell rupture. This method of cell disruption is not scalable or repeatable. A French Press is difficult and time-consuming to clean, and the unit must be cleaned after every sample. Most manufacturers of French Presses have discontinued production, although some outdated units are still in use.
Ultrasonication: This method of cell disruption or cell lysis uses cavitational forces. Often used for very small sample volumes, the cell suspension is sonicated with an ultrasonic probe. Disadvantages of this technique include local high temperatures, resulting in low yields; scalability challenges; and noise. Advantages of this cell disruption technology are low equipment prices and the ability to process small scale volumes.
Freeze-thawing: Subjecting cell suspensions to variable temperatures results in rupture of cell walls. This cell lysis technique is not a very reproducible method, results will vary, and the technique is only suitable for very small samples.
Chemical Cell Lysis: This approach to cell disruption involves adding chemicals that soften and rupture the cell walls. Chemicals can be costly and thus scalability is limited. These chemicals contaminate the preparation which is often undesirable.
Mortar and Pestle: Manually grinding a cell suspension is a laborious process that can take several minutes, making it not scalable and not very repeatable, suitable for small lab samples only.
Media Milling: Contamination by media and temperature control are difficult, but otherwise media milling can be an effective method for disrupting many cell types.