BEE International Blog

Cell disruption is used throughout many industries, from cosmetics and pharmaceuticals to food and biotechnology. As a method or process employed to release biological molecules or other materials from inside a cell, these industries commonly utilize it to study intercellular materials or use the materials located inside a given cell. It can be achieved through a range of methods and technologies, either mechanical or non-mechanical.

The method of cell disruption selected depends on the product, scale and cell type and must obtain the necessary components effectively and reliably without disturbing any of its components. Though high-pressure homogenization is the most common procedure for cell disruption, chemical methods including osmotic lysis, surfactants and chaotropic agents also are prevalent. In this blog, we’ll go into more detail on each of these methods.

Osmotic Lysis

In osmotic lysis (or shock), cells are suspended in a hypotonic extracellular environment, usually a dilute sucrose solution, causing them to take on water, swell and burst. It’s often applied to lyse mammalian blood cells and for RNA extraction, although it is sometimes used to disrupt microbial cells. Due to its low efficiency, however, it is one of the less commonly used chemical methods of cell disruption.

Surfactants

Surfactants (surface acting agents), often called detergents, are compounds that lower surface tension and disrupt the distinct interface between hydrophobic and hydrophilic systems. Their hydrophilic head and hydrophobic tail enable them to insert into and then disperse biological membranes. They are used at fairly low concentrations, and in order to disrupt bacterial cells, they must be used with lysozyme.

Detergents used for disrupting cells are divided three categories depending on their electrical charge: anionic, cationic and non-ionic. All three directly damage the cell wall or membrane, although the best detergents can lyse cells and solubilize proteins. Surfactant cell lysis may be used with ultrasonic processing in order to facilitate disruption.

Surfactants aren’t used for cell disruption more often because they denature proteins in the process and have different impacts on biological systems depending on which detergent is utilized and what its concentration is. No one detergent can be employed for all applications, and some of those selected may disturb downstream processing steps further in the process.

Chaotropic Agents

Chaotropic agents such as urea, guanidine and sodium iodide are capable of bringing some hydrophobic compounds into aqueous solutions by disrupting the structure of water, making it a less hydrophilic environment, and weakening the hydrophobic interactions among solute molecules. Used in very high concentrations, they are like surfactants because they break non-covalent interactions. Unlike surfactants, though, chaotropic agents disrupt the weak interactions between molecules and are sometimes used with detergents to denature and emulsify biological systems.

These techniques are usually only viable at laboratory scale due to increased consumption of energy, chemicals and water and can be very costly for use in large-scale manufacturing. Conditions during the chemical disruption process aren’t always uniform among samples, making it a risky proposition. In addition, some components may cause the denaturation of protein, resulting in a damaged end product.

High-pressure Homogenization

In high-pressure homogenization, a cell disruption method most often used for soft, solid tissues, samples are forced through a narrow space while pressure is applied to them. As the temperature increases, so does the rate of cell disruption. Although they can be used for small batches, high-pressure homogenizers are scalable and can easily adapt to different sample sizes to accommodate increased demand. They not only provide a high level of disruption efficiency but also can be used for the recovery of recombinant proteins.

BEE International: Meeting the Need for Mechanical Cell Disruption

At BEE International, our high pressure homogenizing technology allows you to gently rupture cells without damaging their valuable intracellular materials. You can control the pressure, enabling rupture of a variety of cell types. No harsh chemicals are introduced into the process, and all results are 100 percent scalable to meet your manufacturing needs.

To learn more about cell disruption and how our line of high-pressure homogenizers can help you achieve your goals, please contact us today.

For more information on cell lysis methods and key factors you need to consider when choosing one, download our FREE eBook:

Cell disruption is a critical part of the overall homogenization process, and is used by a number of industries, including pharmaceutical, biotech, cosmetic, and food. But where does cell homogenization fit into the overall process of homogenization? What are the different methods of cell disruption? And most importantly, which method should you use? Continue reading for the answers to all of those questions, as well as more information on BEE International and our suite of high quality homogenization equipment.

Homogenization is also known as micronization, or more simply, particle size reduction. However, before the desired particles within the cell can be released and studied, another step must first take place: cell disruption. Cell disruption is defined as the act of breaking particles apart.

There are numerous forces at play, all of which can be leveraged to disrupt and lyse cells. These forces include impact, shear, turbulence, and cavitation. The type of cell that you wish to disrupt helps drive the decision on which force should be used.

For example, a particle with a harder cell wall, such as yeast, sees the best results with the use of the force of impact. On the other hand, a more delicate particle, such as E. Coli, would benefit from a gentler force such as shear. Other cell types see the best results when utilizing turbulence or cavitation.

When selecting the best homogenization equipment for the task at hand, it is important to have a general understanding of the different types of forces, as well as the forces that should be used for your particular application. This is where BEE International comes in. Our technology allows for the use of multiple forces that can be tweaked and adjusted to yield the highest quality results.

Our equipment offers numerous benefits to the end consumer. Our homogenizers are easy to use, produce a higher yield in less time than our competitors, and allows for the production of repeatable, reliable, and scalable results.

BEE International offers a number of high quality homogenizers for virtually every industry and setting – from a small laboratory or research and development setting all the way up to pilot plants and industrial manufacturing environments. Our homogenizers offer these additional benefits and features as well:

• Ability to control pressure to rupture a variety of cell types
• Ability to rupture cells without damaging the intracellular materials
• Better results in fewer passes

To learn more about the homogenization equipment that BEE International has to offer, please contact us today. We would be more than happy to assist you with your next manufacturing project.

For more information on the different methods of cell lysis and factors you should consider when choosing one, download our FREE eBook:

Cell disruption is the process of releasing molecules or other materials from inside a cell. The action of cell disruption is used in various industries, including pharmaceutical, biotech, cosmetic, food, and drug, and is commonly used by these industries to study intercellular materials or to use the materials that are located inside of a given cell.

Cell disruption can be either a preliminary step to homogenization — the act of micronizing or reducing particle size — , or it can take place simultaneously. In either case, cell disruption can occur via various methods. Continue reading to learn more about the methods for cell disruption.

Bead Method

In the bead method of cell disruption, the cells in question are disrupted via small glass, ceramic, or steel beads mixed with the sample that is suspended in media. In the bead method, the sample and bead mixture is manually agitated by either stirring or shaking.  This is a very common method that is popular in laboratory settings with small amounts of media, and is often used to disrupt yeast cells, as well as various types of animal and plant tissues.

Cryopulverization

The method of cryopulverization is often utilized when dealing with cells with a tougher outer membrane, such as animal connective tissues, cartilage, or seeds. In this method, the material in question is reduced to a very fine powder by impact pulverization at very low (liquid nitrogen) temperatures. Like the bead method mentioned above, cryopulverization is a very manual (and often slow) process.

Microfluidization

Microfluidization is a common method of cell disruption that is often used for proteins, enzymes, and cells, such as E. Coli. In this method, micro channels with fixed geometry are paired with an intensifier pump to achieve high shear rates that easily disrupt even the toughest of cells. While changes in viscosity are often noted with other methods of cell disruption, microfluidization avoids this phenomena, making it an attractive choice for cell disruption.

Nitrogen Decompression

Nitrogen decompression is the method of choice when dealing with more sensitive enzymes, organelles, and cells. In this particular method, large quantities of nitrogen are first dissolved in the cell under high pressure. As the gas pressure is released, the nitrogen escapes, which works to rupture and release the contents of the cell. This technique is used to homogenize cells and tissues, and produces an even dispersion of cells.

To learn more about cell disruption and how our line of high pressure homogenizers can help you achieve your goals, please contact us today.

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

Cell homogenization, also known as cell micronization or cell fractionation, is the action of reducing the particle size of molecules to facilitate even distribution and emulsification of liquids, creams, or other mediums. This process is extremely common, and is used in a number of growing industries: chemical, pharmaceutical, biotech, cosmetic, and food.

In the world of cell homogenization, there are several techniques that are commonly used to achieve this task. All of the methods involve encouraging the cells to lyse, or break apart. Cell homogenization can be achieved through various methods, including mechanical disruption, liquid homogenization, sonication, or manual grinding. Continue reading for a brief overview of each method of cell homogenization.

Mechanical Disruption

Mechanical disruption involves the use of rotating blades. These blades work to grind and disperse cells, and are most effective at homogenizing tissues such as liver.  Rotor-stator homogenizers are one of the best homogenizing tools used in mechanical disruption, and can homogenize samples in the volumes from 0.01 milliliters to up to 20 liters, depending on the type of motor that is used. Sample loss is typically minimal, and small amounts of samples and tissues can easily be homogenized using this method.

Liquid Homogenization

Liquid homogenization is the most widely used cell disruption technique, especially with small volumes and cultured cells. In this method, cells are lysed by the action of being forced through a small space, which acts to shear the cell membranes. There are several types of liquid homogenizers on the market, including Potter-Elvehjem homogenizers, french presses, and the dounce homogenizer.

Sonication

Sonication is a type of physical disruption used to lyse cells. This method uses high frequency sound waves to lyse cells, bacteria, and other types of tissue. The sound waves are delivered via a probe that is immersed in the liquid cell suspension. This method, while common, is often time consuming and is best suited for volumes of less than 100 mL.

Manual Grinding

Manual grinding, while one of the most time consuming methods of cell homogenization, is also the most common. In this method, a mortar and pestle is used to manually grind cells. While not suitable for extremely large volumes, this method is the most effective at breaking apart plant tissue cells.

BEE International offers a wide variety of high pressure homogenizers to meet the needs of virtually any industry. Our technology is well suited for nano emulsions, cell lysis, uniform particle reduction, and other related applications.

Contact us today to learn more about how our line of homogenizers can help with your company’s needs. For more information on cell lysis methods and how to choose the right one, download our FREE eBook:

Cell disruption – the process of releasing biological molecules from inside a cell – is a common method utilized in a variety of manufacturing industries, including pharmaceutical, cosmetic, biotech, and R&D settings.

In order to thoroughly study and analyze the behavior of molecules, the cells that produce the molecules in question must be disrupted. Disruption of cells can be achieved in numerous ways, including cryopulverization, nitrogen decompression, or through using a homogenizer.  Each method comes with varying degrees of complexity, ease, and success.

1. Using Cryopulverization

Cyropulverization is one way of accomplishing cell lysis, and is most often used for cellular samples with a tough outer matrix, such as cartilage, connective tissue, or seeds. In this method, the samples are reduced to a powder through the use of liquid nitrogen and impact pulverization. This method, while successful, is often time consuming and expensive.

2. Nitrogen Decompression

Nitrogen decompression, as the name suggests, uses large quantities of nitrogen to achieve cell lysis. In this particular method, nitrogen is dissolved in the cell under very high pressures. When the pressure is released, the nitrogen violently escapes the cell, causing disruption and breakage of the cell walls. While nitrogen decompression is successful in disrupting certain types of cells – plant cells, bacteria, and other fragile cell types are most receptive to this treatment – it has not been proven to be effective in disrupting cells with tougher outer matrices, such as yeast, spores, and fungus.

3. High Pressure Homogenization

Homogenization is arguably the most widely used method of cell lysis, especially for small volumes of cells and/or cultured cells. This method involves the use of shearing force on the cell. This is achieved by forcing the small cell through an even smaller sized orifice. This removes the outer layer and lyses the cell.

BEE High Pressure Homogenizers

BEE International offers a number of high-quality homogenizers to meet the needs of any industry when it comes to the process of cell lysis and disruption. Our laboratory grade homogenizers include the Nano DeBEE, Micro DeBEE, and Mini DeBee – all three are easy to use, offer consistent and reliable results, and easily scale up for full scale production. In addition, our homogenizers offer maximum experimentation capabilities, the ability to change flow, pressure, shear, and impact settings, and offer high pressures up to 45,000 PSI for maximum effectiveness with cell lysis and disruption.

Contact us today to learn more about how our line of homogenizers can help you meet your production needs. Or for more information on how to achieve the most efficient cell lysis, download our eBook “7 Key Factors to Consider When Choosing a Cell Lysis Method”:

Cell disruption, or cell lysis, is achieved when the cell wall or membrane is ruptured, releasing the contents of the cell. Cell disruption is the first step in many biotechnology applications. This sensitive process needs to be controlled as much as possible; and choosing the correct cell lysis method is essential for preserving the desired intracellular contents.

Cell disruption can be achieved in several ways, including mechanical, enzymatic or chemical lysis. Read on to learn more about chemical methods of cell disruption:

The very simplest form of chemical cell lysis is osmotic lysis. Here, cells are suspended in a hypotonic extracellular environment (often a dilute sucrose solution). This causes the cells to take on water, swell and subsequently burst. Osmotic lysis does not occur in plant cells due to their sturdy cell walls. Organic solvents like alcohols, ether or chloroform can disrupt cells by permeating the cell walls and membranes. These solvents are often used (in combination with shearing forces) with plant cells.

EDTA (ethylenediaminetetraacetic acid) is a chelating agent which can be used to disrupt gram negative microorganisms, since it chelates the cations, leaving holes in the cell walls.

Surfactants (commonly called detergents) disrupt the distinct interface between hydrophobic and hydrophilic systems.  Detergents are used in cell lysis buffers and they help to solubilize membrane proteins and lipids thereby causing the cell to lyse and release its contents. Widely used detergents include Triton and sodium dodecyl sulfate (SDS).

Chaotropic agents, such as urea and guanidine, are also used for cell lysis. They are capable of bringing some hydrophobic compounds into aqueous solutions. They do this by disrupting the structure of water and making it a less hydrophilic environment, and weakening the hydrophobic interactions among solute molecules.

Disadvantages of Chemical Methods of Cell Disruption:

A major drawback in using chemical methods of cell disruption in manufacturing is the cost. Using small amounts of chemicals and enzymes in the R&D laboratory is acceptable; but the cost of using the large volumes required for large-scale production is often not feasible.

Harsh chemicals and detergents can often damage or destroy the contents of the cell if used incorrectly. Lastly, using large volumes of potentially hazardous chemicals creates significant health and safety risks.

The Benefits of Mechanical Cell Disruption with High Pressure Homogenization:

Our high pressure homogenizing technology at BEE International allows you to gently rupture cells without damaging the valuable intracellular materials. You are able to control the pressure, allowing for rupture of a variety of cell types. No harsh chemicals are introduced into the process, and all results are 100% scalable to manufacturing.

For more information on cell disruption, download our FREE eBook: 7 Key Factors to Consider When Choosing a Cell Lysis Method or contact us today!

There exists a small cohort of laboratory techniques that are used regularly by life science researchers and scientists. Among these is cell disruption (also known as cell lysis or rupture), which provides access to versatile intracellular components. Because there are so many ways to disrupt a cell, it is important to understand the broad categorizations, either mechanical or non-mechanical, before narrowing down the mixing method you are interested in using. Below is a descriptive comparison of the two, which can point potential users in the right product direction.

Reference to mechanical cell disruption indicates that the machine uses force to generate a lysate. While some methods, like beadbeating and blending, employ a single force, many others, like grinding and homogenization, incorporate multiple forces, which is advantageous in achieving a higher lysate yield. This mechanical mixing style is advantageous for a number of reasons. Firstly, the use of force instead of detergents and other chemical treatments allows many intracellular proteins to remain intact where they may have otherwise been destroyed. Additionally, mechanical methods are oftentimes better able to process large sample sizes, in terms of both sheer capacity and financial burden.

Non-mechanical disruption, in contrast to mechanical, uses methods other than force to rupture a cell’s wall and/or membrane. For example, enzymatic lysis uses enzymes to disrupt the hydrophilic and hydrophobic bonds in the cell membrane; detergents and chemicals work in a similar fashion. They type of enzyme or chemical will depend on its intended use; for example, proteases like trypsin and collagenase can release individual cells from tissues, while cellulases can yield protoplasts from plant cells. Non-mechanical disruption is most useful for small samples and those which do not have very tough cell walls, as the required time and materials are minimal compared with those of mechanical disruption methods. Additionally, they can target specific cells and/or molecules without leaving contaminants and other unwanted material in the lysate.

BEE International: Trustworthy High-Pressure Homogenizers

Whether you select a mechanical or non-mechanical method of cell disruption will ultimately depend on downstream applications and the cell type being used. Either way, you will be well-served to select a homogenizer that is flexible to meet the various needs of a laboratory. There are plenty of companies on the market to select equipment from; however, the lysate can be of higher quality and more even consistency when run through top-shelf equipment, most frequently in the form of a 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 BEE 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 BEE’s products can maximize your homogenization processes by contacting us today. For more information on 7 factors to consider when choosing a cell lysis method, download our FREE eBook: 

As a scientist/researcher, you can probably think of some lab techniques that have become second nature due to frequent use. However, what if you could make a few tweaks to an already existing technique to make it more efficient? This can translate to savings in both time and money, and importantly, increased yield and quality of lysate. Keep reading to learn about a few tips that will help make your cell rupture technique more efficient.

Decrease Number of Cycles

Although this strategy may only work with some homogenizers, those which may be impacted can see significant results. Most homogenizers require at least 3-4 passes to achieve a consistent and uniform product. However, some higher quality homogenizers can obtain the same results in just 1-2 passes. By decreasing the number of cycles, you can expect to save both reagents/resources and time.

Keep Sample as Cool as Possible

Protein degradation is a common yet unfortunate result of sample mishandling. When the cell membrane is disrupted, proteases that can degrade the molecule of interest, oftentimes a protein, are released. By keeping the sample cool, protease activity can be slowed and even prevented. You can take action by keeping the sample on ice during use, pre-cooling glassware, or working quickly; additionally, it is important to avoid repeated freeze-thawing, which can propagate the degradation process. (1)

Raise Pressure

In a study conducted by Ahmad-Raus et. al., researchers were interested in determining which high pressure homogenization conditions were most important to obtaining a high lysate yield. They found that, specifically among E. coli cells, high pressure (as opposed to medium or low pressure) had a huge impact on the lysate yield. It is therefore advisable to boost the pressure of your own technique for better time and product outcomes. (2)

Use Protease Inhibitors

Because proteases degrade proteins, your protein of interest may be at risk for denaturation if the sample is not handled properly. While cooling can help to slow the actions of proteases, as indicated above, protease inhibitors can completely rid the sample of these irksome molecules. By binding to proteases, protease inhibitors can prevent their destructive actions. As multiple types of proteases exist within a single cell, protein inhibitor cocktails that target many more culprit enzymes are recommended. Conveniently, most science supply companies provide protease inhibitors either individually or as pre-mixed cocktails.

BEE International: The Homogenizer Advantage

One way to implement the above tips is to consider the equipment you are using for your cell disruption technique. As the preferred method, high pressure homogenization will save you precious time while still achieving a high yield of intracellular contents. BEE International Technology manufactures high pressure homogenizers that are trusted by researchers and lab managers around the world. We deliver an array of key benefits - importantly, cell disruption - but also production of nano/micro emulsions and dispersions and lipids and suspensions. These can be used for products across the pharmaceutical, biotechnology, food, and chemical industries.  

Learn more about BEEI homogenizers for cell disruption by contacting us today. For more information on how to pick the cell lysis method that will best suit your needs, download our FREE eBook: 

Cell rupture, a technique used in labs across the world, allows for extraction of intracellular molecules via one of many processes. Chemical and mechanical disruption are two of the more commonly used classes of methods, where enzyme treatment may be employed as a chemical, and high pressure homogenization as a mechanical, procedure. Each may confer significant benefits on the condition of an appropriate match between the researcher’s goal, sample composition, and process capability. Below is an overview of both enzyme and high pressure homogenization procedures in relation to cell rupture, and an analysis on which may be more appropriate for your purposes.

Overview of Processes

Enzyme treatment can be an effective avenue in terms of breaking down cell walls, removing unwanted contaminants, generate protoplasts, and promote DNA isolation by breaking down DNA-binding proteins. The type of enzyme used will depend on its intended use; for example, proteases like trypsin and collagenase can release individual cells from tissue, while cellulases can yield protoplasts from plant cells. One of the major benefits of enzymes is that they can target specific cells and/or molecules without leaving contaminants and other unwanted material in the lysate. (1) High pressure homogenization, in contrast, forces a sample through a narrow space while imparting high amounts of force (e.g. pressure, shear, cavitation, turbulence) on the sample. This allows for effective rupture, particularly among cells whose intracellular components are protected by tough cell walls.

Which Technique Should I Use?

To determine which method will be best for your purposes, start off by identifying three factors about your sample: 1) What type of cell it will be comprised of, 2) The intracellular component(s) of interest, and 3) The process volume. The cell type will determine whether a tough cell wall or thin membrane needs to be penetrated to access the molecule of interest. Additionally, knowledge about the molecule of interest like whether it is secreted from the cell and which enzymes can degrade it will hugely influence your machine selection. Finally, while enzymes are appropriate for small process volumes, high pressure homogenizers are more appropriate for medium and large volumes. Analysis of these three factors as a whole will help you make a more informed and accurate selection.

BEE International: The Homogenizer Advantage

Use of high quality cell lysis equipment will provide numerous benefits for both production laboratories and consumers alike. And there are plenty of companies on the market to select your equipment from. However, the lysate can be of higher quality and more even consistency when run through top-shelf equipment, most frequently in the form of a high pressure homogenizer. BEE International Technology is trusted by researchers and lab managers around the world. We deliver an array of key benefits, such as production of nano/micro emulsions and dispersions and lipids and suspensions; these can be used for applications such as injectables, targeted drug delivery, inhalants, time release, anesthetics, and importantly, vaccinations.

In addition, we have extensive experience in the challenges that our customers face as they transition from concept, through to R&D, clinical trials, all-important FDA approval and finally, to manufacturing.

See how BEE International can help your cell lysis application by contacting us today or if you're just looking for more information on how to choose a cell lysis method, check out our FREE eBook below:

Proteins are indispensable tools for science research and the study of living organisms. Because they are essential to both cell structure and function, much can be learned through protein analysis; these molecules can be accessed through cell rupture, which entails disrupting the cell wall and/or membrane.

In particular, mammalian cell lines are more commonly used than any other cell type because of their easy maintenance, scalability, and genetic modifiability.

Below is a compilation of factors, specific to mammalian cells, that should be considered as you optimize your cell rupture technique.

1. Chosen Technique

Unlike most other cell types with tough exterior walls, the contents of mammalian cells are only separated from the extracellular environment by a thin membrane. Multiple cell rupture techniques exist, such as homogenization, mechanical shearing, freezing, and sonication.

Because primary and culture mammalian cells do not require physical disruption, detergent-based cell lysis is an easy alternative that can be combined with homogenization for optimal results. Detergents work to break apart the lipid bilayer, and homogenizers finalize particle breakage, so that all contents of interest can be accessed.

2. Protein Location & Stabilization

Both cellular location and chemical stability play major roles in determining the best way to approach cell rupture. For example, certain proteins are located in specific organelles. Rupture methods that release the contents from every compartment make it incredibly difficult to achieve high yield of a specific protein of interest.

In contrast, optimization of methods that are able to release contents from specific structures makes the process easier, and enhances product isolation. 

3. Tissue Type

Mammalian organisms are made up of four major tissue types: connective, epithelial, and nerve & muscle. These can be further differentiated into specialized tissues, all of which have unique structures. The tissue type your cell hails from should impact your cell rupture method, as the method may differ with tissues.

According to the Thermo Scientific Cell and Protein Isolation Technical Handbook, physical disruption, such as homogenization, is the best option for cells within tissues, as they are bound tightly within organ-specific matrices. In contrast, primary and cultured mammalian cells simply require some form of enzyme exposure or mechanical dissociation.

BEE International: The Homogenizer Advantage

Whether working with mammalian, yeast, bacterial, or viral cells, it is important to consider the equipment you are using for your cell rupture technique. As the preferred method, high pressure homogenization will save precious time while still achieving a high yield of intracellular contents.

BEE International Technology manufactures high pressure homogenizers that are trusted by researchers and lab managers around the world. We deliver an array of key benefits- importantly, cell rupture- but also production of nano/micro emulsions and dispersions and lipids and suspensions. These can be used for products across the pharmaceutical, biotechnology, food, and chemical industries.  

Learn more about BEEI homogenizers for cell rupture by visiting http://www.beei.com/applications/cell-disruption.