BEE International Blog

Cell lysis is not just a simple and natural process. Conversely, it is both intentional and complex, and critical to researchers involved in a variety of industries. As a researcher, you are probably all-too-familiar with cell lysis and the need for pure lysates. Keep reading for factors that you can adjust to improve the quality of your cell lysate. (1)

1. Cell Type

Regardless of your end goal, if you are lysing cells you are likely looking to extract intracellular contents. However, some cell types lend themselves easier to this process than others. Fungi, yeast, and bacterial have tough cell walls that must first be penetrated; these will therefore confer additional steps compared to an animal cell, whose easy-to-penetrate cell membrane is the only barrier between you and its intracellular contents.

2. Disruption Method

Scientists have the option of using either non-mechanical or mechanical methods of cell disruption. Non-mechanical methods include enzymes, beads, sonication, and detergents; these are advantageous in that they typically require less equipment and only simple reagents. However, non-mechanical methods are best for small sample sizes, and would likely be overwhelmed with a larger sample size. In contrast, mechanical methods such as rotor-stator and valve-type processors, have in common the use of high pressure and better ability to handle large sample sizes. Your laboratory’s specific lysate and product needs will dictate whether a mechanical or non-mechanical method is the best fit.

3. Sample Size

Interestingly, using a sample size that doesn’t match the method can be one of the most significant errors you make. Small sample sizes, commonly used for R&D and independent research, can easily produce a pure intracellular product. However, a larger sample size may pose difficulty in terms of reproducibility and purity of product, particularly when using non-mechanical methods of cell lysis. Homogenization is one of the only methods that achieve a uniform and pure product for both small and large sample sizes.

BEE International: Cell Lysis Homogenizers

Use of a homogenizer to lyse your samples will provide numerous benefits, some of which were touched on above. And there are plenty of homogenizers on the market to choose from. However, the lysate can be of higher quality and more even consistency when run through a top-shelf homogenizer. BEE International Technologies is trusted by researchers around the world for both their laboratory homogenizers and their associated customer support. Cell lysis is just one of a variety of applications for BEEI homogenizers; nano/micro emulsions, lipids, suspensions, and dispersions are also easily achievable. Additionally, the homogenizer processes can be controlled to suit your product, which will allow you to customize to your cell type. And finally, the equipment is easy to use, produces higher yield in less time, and achieves results that are reproducible and scalable.

Learn about how to make your cell lysis protocol more effective by checking out BEE's high pressure homogenizer equipment. Or you can learn more about how to optimize your cell lysis application, download our FREE eBook below:

For the past 545 million years, fungi and yeast have been a critical part of the global ecosystem. Contrary to many current views of them as detrimental to our health, they make a critical contribution in terms of the food web, and importantly, in science research and industry. Specifically, their intracellular proteins can be used to synthesize food and drug components that are vital to human health. However, access to these intracellular proteins can be challenging due to their rigid and protective cell structures. Read below for more on why homogenization is the preferred method for rupture of fungal and yeast cells.

Before we delve into characteristics of yeast and fungi and the logistics of homogenization as a cell disruption method, what we deem as ‘effective’ cell disruption must first be clarified. The goal of cell disruption is access to intracellular proteins for solubilization and/or extraction; therefore, easier access and more usable lysate equates to higher effectiveness in method. An effective disruption method produces a lysate in high quantities and which is highly usable in the application it is destined for.

Although there exist many physical and chemical differences between fungi and yeast, the two species are also incredibly similar. Both classified in the fungi kingdom and each consumes food through decomposition of dead or decaying things. Comparison of the unicellular yeast and a single cell from the multicellular fungus also reveals a strong structural resemblance; this indicates that a similar mode of cell disruption will be undertaken.

Animal cell disruption is typically straightforward with minimal equipment requirements, as they do not contain a cell wall that prevents access to intracellular contents. Yet among plants, fungi, and some protists, this is not the case and will affect which mode of disruption you select. It is recommended that fungi and yeast be exposed to hydrolytic enzymes to break through the cell wall first. Once that process has occurred, homogenization is the preferred method of cell disruption. In previous years, the French press was the machine of choice; however, its low pressure required multiple passes, which can be costly and time-consuming. High-pressure homogenizers, in contrast, can oftentimes disrupt the entire sample in 1-2 passes with enough force to achieve complete disruption, but with enough care that intracellular proteins are not disturbed.

BEEI: Homogenizers That Effectively Disrupt Cells

Researchers who require yeast and/or fungal cell lysates should have a laboratory homogenizer available. On determining which homogenizer will be right for your lab, begin your search with BEE International Technology. They are globally recognized among laboratory managers and researchers for their high quality products and excellent customer support. Cell lysis is just one of a variety of applications for BEEI homogenizers; nano/micro emulsions, lipids, suspensions, and dispersions are also easily achievable. Additionally, the homogenizer processes can be controlled to suit your product, which will allow you to customize to your cell type. And finally, the equipment is easy to use, produces higher yield in less time, and achieves results that are reproducible and scalable.

Learn more about how to effectively lyse your fungal and yeast cells by visiting BEEI here. For more information on how to optimize your cell lysis process, download our FREE eBook below:

Previously, we focused on some common mechanical methods that life science researchers use to achieve cell disruption. These include bead method (a.k.a. beadbetting), sonication, grinding, blenders, freezing, microwave and the use of homogenizers. Now, let us explore some common non-mechanical methods that researchers use to disrupt the cell wall and release the biological molecules within.   

• Chemicals

Often used with plant cells (and sometimes in combination with shearing), organic solvents such as toluene, ether, benzene, methanol, surfactants, and phenylethyl alcohol DMSO can be used to permeate cell walls. Also, EDTA (ethylenediaminetetraacetic acid) can be used to disrupt gram negative microorganisms, since it chelates the cations, which leave holes in the cell walls.

• Enzymes

Enzymes such as beta(1-6) and beta(1-3) glycanases, proteases and mannase can be used to disrupt the cell wall. This method is particularly useful for isolating the cell without the wall (protoplast). Researchers often use EDTA in order to make the peptidoclycan layer accessible.

• Osmotic Lysis

Through the process of osmosis, water can be moved into the cell causing its volume to increase to the point that it bursts. Note that this method can only work with animal cells and protozoa, since they do not have cell walls.

• Electrical Discharges

It is also possible to achieve cell disruption via electrical discharges in mammalian cells, which are cells that are bounded by plasma membranes and, unlike plant cells, have no cell wall. This method allows researchers to examine secretion by exocytosis, which is a process during which the membrane-bounded sphere (intracellular vesicle) shifts to and fuses with the plasma membrane.

• Basic Proteins

Yeast cell walls can be disrupted by using basic proteins, such as protamine.

• BEE Laboratory High Pressure Homogenizers

Many life science researchers are opting to use BEE Laboratory High Pressure Homogenizers in order to achieve cell disruption. This is because our groundbreaking products give researchers the unprecedented ability to control pressure, so that they can rupture a wide variety of cell types – including those with stronger cell walls (e.g. yeast, fungi, etc.). Researchers can also achieve better results, which means a higher yield in fewer passes, which saves time and money, and ensures that results can be scaled to manufacturing.

Learn more about how our laboratory high pressure homogenizers achieve superior cell disruption here. To read more details about each of these methods and about mechnical methods of cell lysis, download our free eBook below!

Cell disruption is a process in which the biological molecules within a cell are released and isolated from the rest of the cell, so they can be analyzed, studied and experimented upon. There are both mechanical and non-mechanical methods of cell disruption. This article looks at some common mechanical methods. A subsequent article will look at some common non-mechanical methods.

• Bead Method (a.k.a. “Beadbeating”)

With the bead method (a.k.a. “beadbeating”), very small beads (0.1-6 mm in diameter) made of glass, ceramic or steel are mixed with a sample that has been suspended in aqueous media (i.e. a solution in which the solvent is water). This process shears open the cell wall, yet in a manner that is gentle enough to ensure that the biological molecules within the cell remain intact.  

• Sonication

Often used for plant and fungal cells, sonication uses ultrasonic homogenizers to induce vibration in a titanium probe that has been immersed in the cell solution. This triggers a process called “cavitation,” which creates very small bubbles that eventually explode and produce shockwaves that ultimately disrupts the cell wall.

• Grinding

It is also possible to achieve cell disruption by grinding via a mortar and pestle. This method is often used with plant samples that have been frozen in liquid nitrogen. Once the cell wall has been disrupted, solvents are added to extract the biological molecules.

• Blenders

The use of blenders (both high speed or Waring) can be used to disrupt cell walls. This is the same process used by centrifugation, which separates or concentrates materials suspended in a liquid medium.

• Freezing

Often used when working with soft plant material and algae, freezing is used to achieve cell disruption via a series of freezing and thawing cycles. Freezing forms ice crystals, which expand upon thawing, and this ultimately causes the cell wall to rupture.  

• Microwave

• Standard Liquid Homogenizers

Homogenizers pump slurry at high pressure (up to 1500 bar) through a valve, which is instantly followed by an expansion through a separate exiting nozzle. Cell disruption is achieved by applying shear forces to the cell membrane.  

• BEE Laboratory High Pressure Homogenizers

A growing number of life science researchers are choosing BEE Laboratory High Pressure Homogenizers, because they represent a radical departure from conventional equipment and provide more experimentation options and capabilities for cell disruption, as well as emulsions, dispersions and liposomes.

Learn more about our groundbreaking laboratory high pressure homogenizers here! To read more details about these cell lysis methods and non-mechnical cell disruption, download our FREE eBook "7 Key Factors to Consider When Choosing a Cell Lysis Method" by clicking the button below!

Cell Rupture / cell disruption / cell lysis, by any name, rupturing cells in a controlled fashion is essential for both research and manufacturing. Some cells, such as yeasts, are much harder to rupture than others. The method chosen for rupturing the cell also has to take into account the nature of the final product- if the cell is being ruptured in order to obtain an active protein, the method chosen has to be gentle enough to not denature or damage the protein.

Enzymatic digestion is often used in the laboratory as a very gentle method of cell rupture. The cell walls of yeast and bacteria are digested with special enzymes and then the unprotected cell can easily be ruptured by a mild osmotic shock. Enzymatic rupture methods are, in general, too expensive to use in large-scale manufacturing.

Chemical methods of rupturing cells are sometimes used. When RNA is the desired product, rupturing cells with a strong chaotropic agent such as urea or guanidine is the approach of choice. These agents denature the proteins, including the many RNA-digesting enzymes present in cells, thus allowing the RNA to be isolated intact. Milder chemical methods, such as using surfactants (e.g. Triton), will allow some active proteins to be isolated. Chemical methods can also be expensive to use in large-scale manufacturing.

Mechanical methods of cell rupture include ball mills, blenders, French presses, homogenizers, and ultrasonic disruption. Ultrasonic disruption is often used in the laboratory with yeast and bacteria, but it is too expensive to use on a larger scale. Blenders and French presses are easy to use in the laboratory, but cannot be readily scaled up for manufacturing. Ball mills are not as popular because the balls need to be removed- an extra step- and usually can only be used once- adding to the cost. Homogenizers are the method of choice for large-scale manufacturing.  

BEE's cell disruption technology comes in both laboratory-sized machines and also in large-scale sizes for manufacturing. The technology is an in-line process that uses cavitation, shear and impact to rupture cells. The forces applied can be precisely adjusted to be as gentle as possible or to be very harsh for more difficult cell types. The sample sizes can vary as well. For example, the Nano DeBEE Laboratory Homogenizer can also handle sample sizes as small as 15 ml.

All aspects of the process can be adjusted- pressure, flow, cavitation, shear, impact and time- allowing the user to fine-tune the method for each application. The precision control of all aspects allows the method to be highly reproducible, and also allows for easy scaling-up. In fact, the Nano DeBEE Laboratory Homogenizer can handle up to 45,000 PSI / 3100 bar for maximum experimentation. The BEE machines are also easy to clean and require little maintenance. 

Don't hesitate to contact us if you have any questions about our cell disruption technology. We have three sizes of machines ideal for any laboratory, and industrial-sized machines for manufacturing. 

For more information on how to achieve efficient and consistent particle size reduction, download our FREE eBook: 

When you're looking for cell disruption equipment for the research laboratory equipment you need a system that is flexible. You're probably aware of the final goal of the research program but since it's research, it's all about trying different things out. If your equipment doesn't let you try things out it's the wrong equipment for the research environment. 

Flexibility

Some cell lysis equipment uses shear. Others use sonic cavitation. Ours uses shear, cavitation and impact- and the user can adjust the relative amount of each force for different applications or to see which procedure works best for a particular application. You may have thought your research team was trying to produce a biological in bacteria and therefore just needed a sonicator, but when they realize the biologic isn't full active unless processed in an eukaryotic system and switch to yeast you'll wish you had bought the BEE homogenizer instead. 

Scalability

Many research laboratory managers don't consider scalability when making purchasing decisions for the research setting. This is one of the reasons the once popular French Press is no longer a viable choice. Because everything is done small-scale in the research laboratory it just isn't a consideration. Let's say your lab started out with bacteria and bought a sonicator. Then they switched to a yeast system. The lab manager refused to buy more expensive equipment, so the team started using an enzymatic lysis method. The purified biologic showed tremendous potential in early experiments. The team proposed to start some clinical trials- which requires scaling up production. However, the costs of scaling up enzymatic lysis of yeast can be prohibitive. The entire research program could be shelved on budgetary grounds before it barely gets started- due to failure to consider "scalability" at the earliest steps. It would be sad if the cure for the cold was shelved because someone decided to not buy a BEE homogenizer. 

As a cell lysis device a BEE homogenizer is fully scalable because all of the parameters are tightly controlled and fully reproducible. After the research team spends weeks changing the parameters on their cell lysis unit to find the optimal method of cell disruption, these parameters can simply be used on a larger scale. All BEE units allow for full control over the pressures and fluid velocities being used during the process. 

Designed for the lab

For the laboratory setting, BEE offers three different sizes of laboratory homogenizers as cell disruption devices. The largest has a flow rate of 250 ml per minute. It can be placed on a standard laboratory bench and is self-cooling so it doesn't have to take up space in the cold room. The smallest unit can process a sample as small as 15 ml, and offers a wide range of operating pressures to allow for experimentation and optimization. It is also self-cooling and is smaller than many desktop centrifuges. The intermediate size can process a sample as small as 15 ml, but can also process up to 15 liters per hour. 

Our cell lysis equipment is the best, whether you want it for the research setting or for large-scale production. Don't hesitate to contact us if you have any questions.