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Top » Product Catalogue » Why / What Is Sonication

What is Sonotransfection?

What is sonoporation and ultrasound-mediated transfection?

Sonoporation is another name for ultrasound-mediated cell membrane permeabilisation or the induction of membrane pores using ultrasound.  When the phenomenon is exploited to facilitate or promote entry of nucleic acid into cells, this is called sonotransfection.   In actual fact, sonoporation can also be exploited in facilitating entry of other agents such as proteins and drugs into living cells and as such it provides the researcher with a valuable tool for applications such as drug screening or metabolic studies.

 

What are microbubbles?

In the field of sonoporation, it is believed that pores are formed in the membrane when ultrasound brings about disturbance in the vicinity of that membrane.  Formation of pores, (sonoporation) can occur at low levels when cells are exposed to ultrasound.  It can however be greatly enhanced by the addition of microbubbles.  These consist of a stabilised lipid shell enclosing a gas (usually a perfluorocarbon gas).  These agents are usually found to be essential when using ultrasound for gene transfection because in their absence transfection efficiency is relatively poor.  Microbubbles, as the name suggests, are small bubbles and they resonate (or vibrate) in the presence of an ultrasonic field.  The optimal resonant frequency of the bubble is a function of the bubble size and so for optimal sonoporation, a match must be achieved between the microbubble size and the ultrasound frequency of the sonoporator device being used.  This is why SONIDEL Limited has designed its own MB101 microbubble to deliver optimal gene transfection with its SP100 sonoporator platform emitting at ultrasound at a frequency of 1 MHz.

 

How do microbubbles work?

Although many studies have been carried out in order to answer this question it appears that the microbubbles work in a combination of ways.   It has been suggested that the bubble will resonate at a specific frequency in an ultrasonic field.  As the intensity or power density of the ultrasound increases, the bubble resonates more violently so that eventually it will collapse in a catastrophic manner.  This effect, if proximal to the cell membrane, will induce rupture of the membrane and this will, in turn, allow reagents into the cell.   Others have shown, using extremely high speed image capture, that the bubble resonates in the ultrasonic field.  In doing so, it forms very strange projections from it surface and it has been suggested that these projections (like needles) transiently rupture the cell membrane thereby permitting transmembrane travel of extracellular substances.

 

How does sonoporation compare with other transfection methods in terms of cell viability?

When all parameters are optimised, sonoporation causes little irreversible cell damage in most cell lines.  Cell membrane recover time has been shown to be less than 10 seconds.  The factors that must be controlled to prevent irreversible cell damage and death are:

- Concentrations of transfection reagent or microbubbles

-      Ultrasound power output

-      Ultrasound application time

-      Ultrasound duty cycle

 

In the published literature, everyone seems to be using different frequencies, which is best?

Yes, this is very true and indeed the field seems quite confused.   This stems, primarily from the lack of commercially-available instrumentation such as has been available to researchers using electroporation over the past number of years.  If one  reviews sonoporation in the scientific literature, one finds that researchers have used different instruments, different ultrasound frequencies, different ultrasound intensities or power densities, different treatment times, different pulse regimes,  different exposure configurations and indeed different microbubble preparations.  It really is extremely confusing.  However, in delving through that body of literature a number of aspects become clear.  One is that 1 MHz seems to yield the most consistent results and has proven most successful for achieving superb membrane permeabilisation with maximal cell viability.   SONIDEL Limited has harvested the best attributes indicated by this extensive body of literature and used those to design its sonoporation platform and associated range of products. 

 

 

For consistent and reproducible results SONIDEL Limited recommends:

-      The SONIDEL SP100 Sonoporation, and

-      SONIDEL STK10 Ultrasound Transfection Kit     

 

Results using the SONIDEL SP100 Sonoporator and STK10 Transfection Kit

 

 

 

 

 SONOPORATION Publications Resource


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