BEE International Blog

A Look at 3 Liquid Homogenization Methods of Cell Lysis

Posted by Deb Shechter on Jul 27, 2015 12:30:00 PM

a-look-at-3-liquid-homogenization-methods-of-cell-lysis

In the liquid homogenization method of cell lysis, samples are pushed through a small space and shear forces are used to disrupt cell walls. This method is common for researchers lysing cultured cells or small volumes.

There are primarily three types of liquid homogenizers used in life science laboratories around the world: Dounce homogenizer, Potter-Elvehjem homogenizer, and French press. But do any of them really get the job done? Below, we take a look at each method.

  • Dounce Homogenizer

Often used for gently lysing single cell suspensions, a Dounce homogenizer uses round glass pestles that are manually forced into a glass tube. While these glass pestles are relatively inexpensive, a major drawback of using a Dounce homogenizer is that the process is typically exceptionally time consuming, and it does not work well with large throughputs – thus making it impossible for researchers to efficiently scale up to manufacturing or clinical trials.

  • Potter-Elvehjem Homogenizer

Often used by researchers who need to disrupt cell walls but not cell tissue, a Potter-Elvehjem homogenizer involves mechanically or manually driving a polytetrafluoroethylene (PTFE) pestle into a conical or rounded-shaped vessel.  Many researchers opt for the manually driven pestles because, while less efficient, they are more affordable. 

  • French Press

Suitable for sample volumes between 40 – 250mL, a French press uses a piston to apply very high pressure to samples, thus forcing them through a very small hole. Compared to a Dounce homogenizer and a Potter-Elvehjem homogenizer, a French press is more efficient (requiring only two passes). However, throughput is low -- thus making it impractical or prohibitive for many researchers who are facing time constraints.

The BEE High Pressure Homogenizers Advantage

Two of the liquid homogenizers described above can ONLY use the mechanical force of impact achieve cell lysis. The French Press additionally employs shears but with little control. However, BEE technology is an in-line process that makes use of three cell disruption forces: shear, cavitation (like sonic mixing) and impact.

As a result, researchers can adjust these forces to be more gentle or harsh, and control the process to rupture a variety of cell types -- including more challenging cells like yeast and fungi – but without damaging tissue or other valuable intracellular materials. The bottom line is better yields in fewer passes, and results that are scaleable to manufacturing and clinical trials.

Learn more about our groundbreaking technology here.

Topics: Liquid Homogenization

7 Cell Lysis Method Factors to Consider

Posted by Deb Shechter on Jul 24, 2015 11:30:00 AM

7-cell-lysis-method-factors-to-considerThere are several factors that life science researchers must keep in mind when choosing the optimal cell lysis method. Below, we highlight 7 key considerations:

1. Force Required 

In terms of strength, not all cell walls are created equal. Some, such as spores, are tougher and therefore require more force to disrupt, while others such as E. Coli are softer and require less force. Force, is related to intensity applied to cells whether it is a stronger concentration of chemical or an increase in mechanical forces exerted on the cell walls.

The ideal method provides the minimum force required to produce the highest yield of lysed cells in the least amount of time. In the laboratory, researchers need the ability to easily experiment and quickly find the ideal force for the cell lysis process.   

2. Volume & Sample Size

Researchers may work with small samples due to lack of product supply, or expensive product cost. But a method that only allows for small samples will prove time consuming when larger batches are required. The ideal laboratory method will allow the scientist to lyse small and medium batches of cells.

3. Scalability

While working with small samples is useful in the lab, time and money can be saved by using a scalable the cell lysis method.  Hopefully, the project at hand will generate enough interest to require larger batches, pilot testing and eventually commercialization. The ideal method will allow for a variety of sample sizes and must be scalable so that the results achieved in the laboratory can be duplicated in pilot and manufacturing.

4. Sample Variety

When purchasing equipment, your dollar will always go further with a versatile selection.  The ideal method can be used for a wide variety of cells as well as suspensions, emulsions, dispersions, liposomes, etc. 

It is also necessary to adjust to the number of different samples that will be disrupted concurrently. Failure to adjust the equipment and technology can result in diminished processing speed. Failure to clean equipment can also lead to low yield.  

5. Efficiency

Efficiency considerations are important to ensure the feasibility of scalability and to produce a high yield of the cellular material sought with the greatest ease. The ideal method will produce the highest percentage of lysed cells in the shortest amount of time. In the laboratory the amount of time and cost to lyse a batch of cells may not be critical. But in manufacturing it may make the difference between whether or not a product will be profitable.  Some cell lysis methods have an inherent trade off  between disruption efficiency and ensuring that sub-cellular components remain intact. This may be related to the type of cell being disrupted.

6. Product Stability

Once cell lysis has occurred, researchers must be prepared to protect the extracted proteins so that they can be isolated, examined, studied and experimented upon. The ideal method will maintain product stability to avoid denaturing of cells. Methods that cannot control factors such as normal biological processes, temperature change and oxidation will produce diminished yields.

7. Meeting Sanitary Standards

The Pharmaceutical and Biotech industries must comply with the highest Sanitary Standards  ...applications must conform to the stringent FDA and cGMP sanitary requirements. From welded components to the completed system care must be given to regulations are met and the safest, most effective pharmaceuticals and biotech products are produced. The ideal system must be easy to clean and designed for sanitary applications.

BEE Laboratory High Pressure Homogenizers

BEE Laboratory High Pressure Homogenizers are the preferred choice among research laboratory managers worldwide, because they are designed to easily support the widest range of experimentation. Also since cell lysis is one of the most common uses for this equipment, the critical factors described in this article have all been addressed.  

BEE systems offer the widest range of force (operating pressure) from 5 kpsi to 45 kpsi compared to other systems stop at 18, 23 or 30 kpsi. Increasing or decreasing force is as easy as turning a dial.

BEE technology is an in-line mixing process so one laboratory system can be used for sample sizes ranging from 20 ml to 25 liters per hour.

Not only do BEE laboratory systems produce a high yield of lysed cells for all cell types, but the ability to experiment with many options also produces the ideal process for a given product.

BEE high pressure homogenizing technology does generate heat which can cause damage to the precious cellular material. However our cell lysis systems include custom designed heat exchangers to maintain product temperature and avoid denaturing of cell materials.

They are also easy to clean and maintain, which lowers the risk of cross-contamination. Just as importantly, the technology used in our equipment is designed to deliver high yield with fewer passes, which is both time and cost efficient, and allows researchers to scale up to conduct clinical trials.

BEE systems are designed for sanitary applications and sanitary options are available to meet industry requirements. BEE systems offer Clean in Place (CIP) and are suitable for Clean Room environments.

Learn more about our laboratory high pressure homogenizers here.

Topics: cell lysis

A Look at 4 Common Downstream Applications Following Cell Lysis

Posted by Deb Shechter on Jul 23, 2015 12:30:00 PM

a-look-at-4-common-downstream-applications-following-cell-lysisAfter scientists disrupt cell membranes through the process of cell lysis and remove the biological material within, they carry out various downstream applications to fulfill their research goals and objectives. This article looks at 4 common downstream applications following cell lysis: protein purification, proteomics, Western blotting and immunoprecipitation.

Protein Purification

Prevalent in the biotech and pharmaceutical industries, protein purification involves several processes and specific tools designed to isolate one or a small number of proteins from the cell, so that the protein(s) function, structure and interaction can be studied.

Proteomics   

Often used in disease treatment research, drug discovery and proteogenomics (studies based on proteomic data in order to improve gene annotations), proteomics is the large scale study of the full set of proteins produced by a system or organism; and in particular, their functions and structures. During proteomics, researchers purify and analyze proteins using various tools (e.g. electrophoresis systems, mass spectrophotometry systems, etc.). Proper tool selection depends on various factors, including the type of cell, the biological system of interest, and time, size and budget limitations faced by the laboratory. 

Western blotting

Western blotting (a.k.a. immunoblotting) is a common downstream application that uses antibodies to help researchers identify specific proteins from a sample. There are three aspects of the western blotting process: 1) separating the extracted proteins based on size; 2) transferring the proteins to a solid support such as an enzyme or fluorescent dye; 3) visualizing the target protein via chemiluminescent or chemifluorescent detection reagents, or by using the fluorescent tag.

Immunoprecipitation 

Immunoprecipitation – often referred to in research literature by the acronym IP – is a process that leverages the antigen-antibody reaction principle in order to enable the purification or a protein (or a protein complex), so that researchers can examine physical characteristics or quantity. There are typically four steps in the process: 1) the proteins extracted via cell lysis are suitably precipitated; 2) the immune complex is captured on a solid support upon which Protein G or Protein B has been immobilized; 3) elements that bind to the immune complex are removed via elution from the support; 4) researchers further analyze the immunoprecipited proteins as per their plans.  

BEE International High Pressure Homogenizers: Designed for Downstream Applications

All of our high pressure homogenizers are designed to support a wide range of downstream applications, including (but not limited) to those described in this article. We accomplish this  by producing the highest yield cell lysis in the shortest amount of time. Each of the systems in our line-up consistently produce the same results, have a reputation for lasting reliability, scale up to pilot and clinical trial settings, and use in-line processes to reduce costs by achieving better results in less time.  

Learn more about our groundbreaking high pressure homogenizers here.

Topics: cell lysis

An Overview of Subcellular Fractionation

Posted by Deb Shechter on Jul 22, 2015 11:30:00 AM

an-overview-of-subcellular-fractionationSubcellular fractionation is the process separating the membrane-bound organelles within eukaryotic cells, which are found in the kingdoms Protista, Plantae, Fungi and Animalia. All eukaryotic cells share certain general features, including:

  • They all have a nucleus.

  • They are 10x larger than prokaryotic cells (which do not have an organized nucleus).

  • They are enclosed by a plasma membrane.  

  • Their cytoplasm is made of cytosol and ribosomes.

  • They have an internal cytoskeleton.

  • Their extra-cellular matrix is comprised of proteins and glycoproteins.

  • Locomotion is achieved through flagella or cilia.

Reasons for Subcellular Fractionation  

There are a couple of key reasons why life science researchers need to conduct subcellular fractionation. The first is to learn more about a protein’s function and where it resides. The second is to improve the results of immunoprecipitations (such as removing unwanted proteins).

Protocol Selection

Prior to subcellular fractionation, researchers must determine what aspect of the organelles within eukaryotic cells they wish to study, such as protein activity, organelle morphology, protein composition, and so on. There are several protocols available to assist researchers once they have determined their research goals.   

Subcellular Fractionation Protocol Steps

Generally, there are 4 subcellular fractionation protocol steps as follows:

  • Step 1: Cell Lysis

The correct cell lysis method depends on a number of factors, including protein type, the organelle within the eukaryotic cells that researchers want to examine, and various downstream applications (e.g. protein purification, proteomics, X-ray crystallography, Western blotting, immunoprecipitation, etc.).

  • Step 2: Subcellular Fractionation

Next, researchers use centrifuging in a high viscosity media (such as sucrose, glycerol or Percoll) in order to achieve subcellular fractionation. A number of factors must be taken into consideration here to ensure that the process is efficient, cost-effective, and yields the best possible results.  

  • Step 3: Collect Fractions

Researchers then collect fractions by a process of gently pipetting through the high viscosity media.

  • Step 4: Assess Results

Lastly, researchers verify and assess their results by (for example) running fractions on a Western blot.

BEE International’s Proprietary Technology

BEE International’s proprietary technology is designed to utilize all available mechanical forces to help researchers break particles apart – unlike other technologies, which apply just one mechanical force to mix a product.  As a result, researchers can achieve the ideal process for producing the highest yield of viable lysed cells in the shortest amount of time, increasing manufacturing efficiency, and reducing costs!

Learn more about our groundbreaking, proprietary technology here.

What to Consider in Designing a Protein Purification Buffer

Posted by Deb Shechter on Jul 21, 2015 11:30:00 AM

what-to-consider-in-designing-a-protein-purification-bufferDuring cell lysis, the extracted proteins can become denatured or damaged. They can also separate from the assay solution, or be contaminated by exposure to lipids, DNA, and other irrelevant cell components. To avoid this from happening, researchers use a buffer solution, which helps ensure the integrity and stability of the proteins. Below, we look at some key factors to consider in designing a suitable protein purification buffer.  

  • Buffer System

Buffer systems are designed to resist change in the assay solution’s pH. These systems have significant amounts of weak acid and its conjugate base (formed when the acid donates a proton), or a weak base and its conjugate acid added to the assay solution. The goal is to create a buffer that has a pKa value within a single pH unit of the optimal pH.

  • pH Level

Speaking of pH: researchers need to identify the suitable pH level for the protein of interest. Many researchers aim for pH 7.4, because this considered the healthiest pH level for blood (specifically, between 7.35pH – 7.45pH), and therefore aligns with ideal biological conditions.

  • Additives & Agents

Researchers also need to add various additives or agents to the buffer in order to enhance protein solubility and stability. Examples include bovine serum albumin (a.k.a. BSA or Fraction V), which derives from cows, or small amounts of citrate or detergents. Researchers may also need to add viscosity, which can be done by adding an agent like polyethylene glycol (a polyether compound).

  • Salt

Researchers often use salt to both enhance protein solubility and stability, as well as align with ideal or desired biological conditions. Often, the salt concentration must be modified (via dialysis of the protein in a new buffer) while protein purification is taking place, in order to avoid nonspecific interactions, and to detect ionic interactions.   

  • Reducing Agents

If oxidation is a risk factor, then researchers may need to add reducing agents to their protein purification bugger, such as DTT, TCEP, 2-mercaptoethanol etc. Many researchers prefer TCEP because it acts in a broader range of pH, and because it’s very stable. However, it is quite costly, which makes it prohibitive in some research programs.   

The Bottom-Line: It’s All About Yield!

When it comes to cell lysis and examining isolated proteins, the objective that all life science researchers have – and especially laboratory managers -- is the same, regardless of whether they work in the food industry, biotech, pharmaceutical field, or anywhere else: it’s all about yield!

At Bee International, all of our laboratory homogenizers are built for reliability, and designed to produce repeatable and scalable results, so that researchers can maximize yield in the fewest passes possible – and ultimately scale up to manufacturing or clinical trial as rapidly, reliably and cost-effectively as possible. Learn more here.

3 Best Practices for Preventing Contamination in Life Science Laboratories

Posted by Deb Shechter on Jul 16, 2015 12:30:00 PM

 

3-best-practices-for-preventing-contamination-in-life-science-laboratoriesBiological contamination is a constant threat in life science laboratories, and there is frankly no “bulletproof” way to prevent it. Yet with that being said, there are certainly proven ways to help minimize the risk of contamination, which not only saves time and money, but even more importantly, helps keep laboratory personnel safe.

With this in mind, here are 3 best practices to maintain the integrity of cell cultures, and promote a safe laboratory environment: 

1. Use Appropriate Lab Design

It is important that the lab has a specific area that is only used for cell culture. This area should be as far away as is practical from high-traffic areas, and it should only be accessed by authorized personnel. HVAC units, sinks, and other items or equipment should also be placed accordingly so as to minimize accidents or contamination. This is because the back splash from sinks can be a source of microbial contamination, and poorly-placed HVAC units can blow mold spores into the cell culture area.

2. Use Correct Culturing Procedures

All lab personnel should be trained to follow correct culturing procedure, which includes proper aseptic techniques. For example, it is vital to work with one cell at a time in order to avoid unintentional switching of cell lines, which can ultimately lead to flawed and unreliable data. It is also important to test for mycoplasma on a monthly basis, as well as to avoid routine antibiotics, as these can hide the existence of underlying contamination. And of course, Good Pipetting Practice (GPP) is essential to support sample integrity and accuracy. 

3. Use Suitable Cleaning Procedures 

It is necessary to implement standardized laboratory cleaning and disinfecting processes, and to ensure that they pertain to both work and non-work surfaces – since such surfaces rapidly collect potential contaminants such as dust. A sufficiently-stocked Biological Safety Cabinet (BSC) is also critical, and it should be placed in an area that is accessible, ensures appropriate air flow and filtration, and of course, prevents contamination. 

Also keep in mind that the BSC is exposed to microorganisms every time the door is opened. As such, advanced incubator design is required. For example, some incubator designs feature HEPA filtration that establishes ISO 5 cleanroom conditions within five minutes of the door opening. Other designs use a 100% pure copper internal chamber and components, and use high temperature decontamination. And there are also designs that feature CO2 sensors and humidity control.

The Bottom-Line

Although, as noted above, it is impossible to 100% prevent the possibility of contamination in laboratory science environments, there certainly are proven ways to mitigate the risk. These include appropriate lab design, correct culturing procedures, and suitable cleaning procedures. 

BEE International Laboratory High Pressure Homogenizers

At BEE International, our laboratory high pressure homogenizers are designed for sanitary applications, easy to clean and maintain, which make them an essential part of an overall system to help reduce the risk of contamination in life science laboratories -- which saves money and time, and helps keep laboratory personnel safe and out of harm’s way.

Learn more about our laboratory high pressure homogenizers here

Topics: Biotechnology, Contamination

A Look at Different Types of Cell Walls

Posted by Deb Shechter on Jul 13, 2015 12:30:00 PM

a-look-at-different-types-of-cell-wallsCell lysis is the process of disrupting cell walls in order to extract and isolate proteins, so they can be analyzed and experimented upon by life science researchers. Below, we provide an overview of different cell wall types.

  1. Plants

Plant cell walls have a primary membrane, and may have a secondary membrane as well. The primary membrane is made mostly of polysaccharides cellulose, pectin and hemicellulose. It is flexible, which allows the plant to grow properly, yet at the same time it is sturdy enough to establish turgor pressure, which is necessary to support the plant’s stability (i.e. water inside the cell presses up against the cell wall). The secondary membrane, if present, contains lignins, which are complex natural polymers, and one of the main classes of structural materials in the support tissues of vascular plants (as well as some algae). Lignins play an important role in making the cells waterproof and supporting xylem.

  1. Algae

Algal cell walls are made primarily of polysaccharides, which are long chains of polymeric carbohydrate molecules composed linked by glycosidic bonds. They can also contain cellulose, mannan or xylan. In addition, some algal cell walls -- such as those of brown algae -- contain alginic acid that is capable of absorbing water, and form a kind of gum that is used by researchers in the cosmetics and food industry.  

  1. Bacteria

Bacteria cells walls are comprised mainly of peptidoglycan, which is a polymer of amino acids and sugars. The result is a structure that looks something like a chain link fence, which is strong enough to support the cell, yet porous enough to allow particle movement. There are two types of bacteria cell walls: Gram-positive and Gram-negative. Gram staining is used to distinguish between them.  

  1. Archea

Archea cells walls are still being researched and not as fully understood as other types of cells walls. However, what we know thus far is that many archaea contain pseudo-peptidoglycan, which is created from the assembly of surface-layer proteins (S-layers).

  1. Fungi

Fungal cells walls are made of the polysaccharide chitin, which is somewhat similar to cellulose, but contains acetyl-amin (nitrogen) groups rather than hydroxyl-groups. True fungi cells walls also contain glucans (glucose polymers) and proteins, which support cell wall synthesis and lysis.  

BEE Technology: Designed to Disrupt Various Types of Cell Walls

At BEE international, the technology we use to design our equipment allows researchers to control and modify pressure, so that they can rupture a variety of cell types – including more challenging cells such as fungi – but without damaging the intracellular materials. This results are a higher yield with fewer passes, and results that can be scaled up to manufacturing. Learn more about our groundbreaking technology here.

Topics: Cell Walls

An Overview of Non-Mechanical Methods of Cell Disruption

Posted by Deb Shechter on Jul 10, 2015 12:30:00 PM

 

an-overview-of-non-mechanical-methods-of-cell-disruption

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.

Topics: cell disruption

An Overview of Mechanical Methods of Cell Disruption

Posted by Deb Shechter on Jul 9, 2015 1:00:00 PM


an-overview-of-mechanical-methods-of-cell-disruptionCell 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

Microwave (along with autoclave and other high temperature methods) are used to disrupt the bonds within cell walls, and also to denature proteins. This is a somewhat risky method, as the excess heat can quickly damage the rest of the cell.
  • 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!



Topics: cell disruption

Cell Lysis Snapshot: Sonication

Posted by Deb Shechter on Jul 9, 2015 12:30:00 PM

cell-lysis-snapshot-sonicationThere are several methods used in life science laboratories to break open cells (a.k.a. achieve cell lysis) and access the materials within (e.g. DNA, proteins, organelles, proteins, DNA, mRNA, etc.). One such method is sonication. 

What is Sonication?

Sonication breaks open cells via the process of sonochemistry. A metal probe is immersed in the sample containing the cells. A power source attached to the probe, which generates sound energy typically in the 20-50kHz range.

This sound energy is then converted via the ultrasonic probe into mechanical energy, which causes an implosion of tiny bubbles in the sample. This is known as “cavitation”, and is what ultimately triggers cell rupture and enables cell lysis.

Note that another, somewhat less common application of sonication is by using an ultrasonic bath instead of an ultrasonic probe. 

Limitations of Sonication

While sonication is a common method for achieving cell lysis, it is not without drawbacks and limitations; some of which are significant. Here is a snapshot of some of these weaknesses: 

  • Accessibility: Because it unleashes a violent implosion of “bubbles” in the cell culture sample – increased temperatures can result in denaturing proteins. As such, it is not suitable for less resistant cells. 
  • Efficiency: Sonication may require numerous short runs for larger samples, and therefore may be inefficient.
  • Costs: Because of the added time investment and the risk damaging the cell wall, relative to other methods sonication is not a cost-effective process, and is not an in-line process. 
  • Yield: Sonication can lead to variations in yield, due to the nature of random vibrations. This can impede the ability to develop a consistent manufacturing protocol.
  • Contamination: Sonication generates free radicals, which can react with other molecules and cause contamination. 
  • Configuration: Sonication equipment must be optimized (time, power) for each cell type. This can be inefficient and lead to added time and costs.  

BEE High Pressure Homogenizers

Researchers in life science laboratories who want to avoid these drawbacks and limitations of sonication are invited to learn more about BEE International’s High Pressure Homogenizers, which are suitable for different cell disruption strategies (e.g. gentle, harsh). They are also efficient and deliver high yields in less time, and scale to allow researchers to go from small samples to larger clinical trials, but without impeding the ability to reproduce results.  

Learn more about BEE International’s High Pressure Homogenizers by clicking here.

Topics: cell lysis, Sonication

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