Sonication Application of Sonication Principle of Ultra Sonication Sonication Methods Probe Sonicator Assembly

1. 1
Probe Sonicator
Dr. Anil Pethe
Shobhaben Pratapbhai Patel School of Pharmacy & Technology Management,
SVKM’S NMIMS, Mumbai

2.  Sonication
 Application of Sonication
 Principle of Ultra Sonication
 Sonication Methods
 Probe Sonicator Assembly
Contents

3.  Sonication is the act of applying sound energy to agitate particles in a sample,
for various purposes.
 Ultrasonic frequencies (>20 kHz) are usually used, leading to the process also
being known as ultrasonication or ultra-sonication.
 In the laboratory, it is usually applied using an ultrasonic bath or an ultrasonic
probe, colloquially known as a sonicator.
 Sonication is the mechanism used in ultrasonic cleaning—loosening particles
adhering to surfaces. In addition to laboratory science applications, sonicating
baths have applications including cleaning objects such
as spectacles and jewelry.
 Sonication is commonly used in nanotechnology for evenly dispersing
nanoparticles in liquids. Additionally, it is used to break up aggregates of
micron-sized colloidal particles.
Sonication

4. Sound frequency ranges
Ultra-Sonication

5.  It is applied in pharmaceutical, cosmetic, water, food, ink, paint, coating, wood
treatment, metalworking, nanocomposite, pesticide, fuel, wood product and
many other industries.
 Sonication can be used for the production of nanoparticles, such
as nanoemulsions, nanocrystals, liposomes and wax emulsions.
 For wastewater purification, degassing, extraction of plant oil, extraction of
anthocyanins and antioxidants, production of biofuels, crude oil
desulphurization, cell disruption, polymer and epoxy processing, adhesive
thinning, and many other processes.
 Sonication can be used to speed dissolution, by breaking intermolecular
interactions. It is especially useful when it is not possible to stir the sample, as
with tubes.
 It may also be used to provide the energy for certain chemical reactions to
proceed.
Application of Sonication

6.  In biological applications, sonication may be sufficient to disrupt or deactivate a
biological material. For example, sonication is often used to disrupt cell
membranes and release cellular contents. This process is called sonoporation.
 Small unilamellar vesicles (SUVs) can be made by sonication of a dispersion of
large multilamellar vesicles (LMVs).
 Sonication is also used to fragment molecules of DNA, in which the DNA
subjected to brief periods of sonication is sheared into smaller fragments.
 Sonication can also be used to initiate crystallisation processes and even
control polymorphic crystallisations. It is used to intervene in anti-solvent
precipitations (crystallisation) to aid mixing and isolate small crystals.
 Sonication can be used to remove dissolved gases from liquids (degassing) by
sonicating the liquid while it is under a vacuum. This is an alternative to
the freeze-pump-thaw and sparging methods
Application of Sonication

7.  The desired effects from the ultrasonication of liquids –
including homogenization, dispersing, deagglomeration,
milling, emulsification, extraction, lysis, disintegration and
sonochemical effects – are caused by cavitation.
 By introducing high power ultrasound into a liquid
medium, the sound waves are transmitted in the fluid and
create alternating high-pressure (compression) and low-
pressure (rarefaction) cycles, with rates depending on the
frequency.
Principle of Ultra- Sonication
 During the low-pressure cycle, high-intensity ultrasonic waves create small vacuum
bubbles or voids in the liquid. When the bubbles attain a volume at which they can no
longer absorb energy, they collapse violently during a high-pressure cycle.
 This phenomenon is termed cavitation. During the implosion very high temperatures
(approx. 5,000K) and pressures (approx. 2,000atm) are reached locally. The implosion of
the cavitation bubble also results in liquid jets of up to 280m/s velocity.

8. Sound propagation in a liquid showing cavitation bubble formation and collapse
Principle of Ultra- Sonication

9. Sonication Methods
 DIRECT Sonication Method
inserting a probe directly into a sample vessel
 INDIRECT Sonication Method
eliminates the need for a probe to come in contact with your sample.

10.  DIRECT Sonication (inserting a probe directly into a sample vessel)
is the most common way to process a sample.
 Energy is transmitted from the probe directly into the sample
with high intensity and the sample is processed quickly.
 The diameter of the probe’s tip dictates the liquid volume that
can be effectively processed.
 Smaller tip diameters (Microtip probes) deliver high intensity
sonication and the energy is focused within a small, concentrated
area.
 Larger tip diameters can process larger volumes, but offer lower
intensity.
 Boosters and High Gain horns can be used to increase the output
of large diameter probes.
 Probes are offered with either replaceable or solid tips and are
made from titanium.
Direct Sonication Method

11.  INDIRECT Sonication eliminates the need for a probe to
come in contact with your sample.
 This technique is often described as a high intensity
ultrasonic bath.
 The ultrasonic energy is transmitted from the horn, up
through the water and into a vessel or multiple sample
tubes.
 Indirect sonication is most effective for very small samples
because foaming and sample loss are eliminated.
 Pathogenic or sterile samples are ideal for this method
because cross contamination are prevented.
 The Cup Horn and Microplate Horn deliver indirect
sonication and are ideal for many high throughput
applications.
Indirect Sonication Method

12. Simplest – ultrasonic baths Ultrasonic probe system
Probe Sonicator Equipment

13. A sonicator has three main components:
a generator, a transducer, and a probe.
The generator transforms the input electrical
power to an electrical signal that drives the
transducer.
The transducer converts the electrical signal into a
vibration. This vibration is amplified as a
longitudinal vibration in the probe tip, causing the
sample to cavitate.
Cavitation creates the ultrasound energy, which
causes the sample to disrupt and break down into
smaller particles
Component of Sonicator

14. Probe Sonicator Accessories
 Horns (also known as probes)
 Direct Horn Options
 High Throughput Horns
 Indirect Horn Options
 Sound Enclosure
 Flocells
 Chillers
 General Accessories

15.  Horns (also known as probes) are made from titanium and machined to specific sizes
and shapes.
 When driven at their resonant frequency, they expand and contract longitudinally. This
mechanical vibration is amplified and transmitted down the length of the probe.
 In liquid, the probe causes cavitation which constitutes the main mechanism for
sample processing.
 Choosing the appropriate horn is extremely important.
 The sample volume to be processed is directly related to the tip diameter. Smaller tip
diameters (Microtip probes) deliver high intensity sonication, but the energy is focused
within a small, concentrated area.
 Larger tip diameters can process larger volumes, but offer lower intensity.
 Probes are offered with replaceable or solid tips.
 Probe tips will pit or erode over time and require replacement.
 Replaceable tip probes are used with aqueous samples only.
 In addition to aqueous samples, Solid probes can be used with organic solvents,
alcohols and low surface tension liquids.
Direct Horn Options

16. Replacement Tips for Standard Probes
 Standard ½", ¾" and 1" horns have replaceable
tips.
 During normal use, tips erode and become less
effective over time.
 These worn tips can be easily removed and
replaced.

17. Microtip Probes
 Microtips are thin, high intensity probes which are designed for processing
small sample volumes.
 Microtips screw into the threaded end of the standard ½" probe

18. Extenders
 Standard probes may not be long enough
to fit down into certain long necked
vessels.
 Extender probes attach to standard horns
of the same tip diameter and extend the
length of the horn assembly.
 Extenders are available in 5" and 10"
lengths with either solid, or replaceable
tips.
 Extenders offer the same processing
volume and amplitude of their
corresponding standard horn.

19. Booster horns
 Increase the intensity of standard ¾" and 1"
horns.
 Boosters attach between the converter and
horn to increase amplitude by the gain ratio
indicated below.
High gain horns
 High gain horns (also known as high
intensity horns) offer double the amplitude
of standard ¾" and 1" horns.
 High gain horns attach directly to the
converter.

20. The 4 Tip Horn
 The 4 Tip Horn enables 4 samples to be processed
simultaneously.
 This horn offers high intensity and is effective for cell
disruption, mixing, homogenization and many other
applications.
 Tip diameter is 1/8" and the space between each tip is 1.05".
 The 4 Tip Horn can process 1-15ml sample volumes and is
made to fit into both 1.5ml and 15ml tubes.
 When processing small volumes with high intensity, samples
will heat up quickly. In addition to using the pulse mode, a
CoolRack tube cooling module is highly recommended.
CoolRack accessories work well with the 4 Tip Horn.
High Throughput Horns

21. The 24 Tip Horn
 The 24 Tip Horn processes each well of a
24 well plate simultaneously.
 This horn is effective for cell disruption,
mixing, dissolution and many other
applications.
 The 24 Tip Horn can be mounted inside
the Sound Enclosure to reduce the noise
level generated by sonication.
 Alternatively, a Heavy Duty Stand is
available which allows precise
adjustment of the horn in and out of the
microplate.

22. Dual Horn
 The Dual Horn allows a single Sonicator unit to process two
samples simultaneously.
 The rectangular-shaped horn doubles the unit’s output, and
enables two probes to vibrate with the same intensity as a
single probe.
 The distance from center to center of each probe is 4.5".
 ¾" solid tip probes are included with the Dual Horn but ½"
or 1" probes may also be used.
 The Dual Horn is capable of withstanding the rigors and
harsh chemicals of environmental testing labs.

23. A Cup Horn
 A cup horns offers indirect sonication and functions as a
high intensity ultrasonic water bath. Multiple samples can
be processed in sealed tubes eliminating cross
contamination or aerosol issues.
 The horn is mounted within an acrylic cup and the cup is
filled with water. Sample tubes are placed in a rack at a fixed
distance above the ultrasonic horn. Cavitation is produced in
the water, processing the samples within the tubes.
 Sonication generates heat so ports for cooling are located
on each side of the cup.
 The Chiller is recommended for maintaining both the water
temperature and water level within the Cup Horn.
Indirect Horn Options

24. Microplate Horn
 Similar to a Cup Horn, but larger, the Microplate Horn
is an indirect sonication device capable of processing
an entire 96 well microtiter plate or many microtubes
at one time.
 Simply place your samples within the water-filled
reservoir and the sonic energy is transferred into each
individual well or tube.
 The Horn is equipped with a clear acrylic collar to
contain the liquid media within the reservoir.
 This allows the user to process deep well microplates
or other tall vessels. Standard microtiter plates or PCR
tubes require a smaller volume of liquid for sonication.
For these applications, the clear acrylic collar may be
removed and the lower, gray collar will allow for easier
access to the samples.
Indirect Horn Options

25.  Sonicators are extremely loud devices and will cause
discomfort to the user and anyone nearby.
 The Sound Enclosure reduces noise by approximately 20
dBa and is made to work with all accessories
 In addition to reducing noise, the Sound Enclosure has an
internal support rod and converter mounting system.
 Any probe or horn will be held safely and securely inside
the unit.
 Two ports are located on either side of the enclosure for
coolant tubing or a temperature monitoring probe. The
interior walls are lined with acoustical foam and the door
has a window so experiments can be visually monitored.
Sound Enclosure

26.  Flocells offer inline or continuous, large volume, batch
sample processing.
 Flocells are ideal for mixing and dispersing applications.
Batch volumes can be re-circulated through the system
multiple times if increased sonication time is needed.
 Multiple units can be used in series to reduce
processing time and/or maintain an even higher flow
rate.
 The liquid sample is pumped into the Flocell through
the inlet at the bottom of the unit. As the sample
passes through the cavitation field, it is processed. The
processed liquid exits the unit through an outlet port.
The degree of processing can be controlled by adjusting
the intensity of sonication as well as flow rate.
Flowcells

27. Recirculating Chillers
 Sonication generates heat which may be
detrimental to some applications.
 Attempting to control temperature with ice and/or
repeatedly changing out water is tedious and no
longer necessary.
 Automating the sample cooling process with 2
chiller options are available
 Quick-connect tubing and fittings (ordered
separately) attach the chiller to the ports on the
cup horn or microplate horn.
 When used in conjunction with the pulsed
sonication mode, your desired water temperature
will be maintained.
Chillers