Microbubble preparation

and characterisation for

the biomedical
applications

Ketan Pancholi,
School of Engineering, Robert Gordon University, Schoolhill, Aberdeen ,AB10 1FR
 T-junction

ng R egion
Fo cussi

20µm

• T-junction1,2 three dimensional aligned and easy to

assemble.
• For each oil of given viscosity , the effect of varying the gas
pressure (Pg) from 0.64 KPa to 17 KPa was determined for
differentM.Edirisinghe
1 K.Pancholi, liquid flow rates
(2007) (Ql) from
Microbubbling 1.66
of viscous x 10
media, -10
toPhysical
American 8.33 Society,
x 10-8
March Meeting , Denver, USA
m3/s.   patent. PCT filed on 28/03/08 (PCT/GB2008/001050)
2 *International
Experimental Set-up
High pressure withstanding
T-junction enclosed in
polymer block with guiding
holes to self align tubes on
its own axis. Total set up is
shown including high speed
camera, stainless steel
syringe able to withstand
50MPa pressure. •First air thread enters liquid
column downstream to establish
two phase flow*
•Air column reduces effective area
for liquid to flow
•This increases pressure at air
liquid interface
as liquid is supplied at constant
flow rate by pump to maintain flow
rate.
•A sharp increase in liquid pressure
normal
*K.Pancholi, E.Stride, M.Edirisinghe (2008) Dynamics of bubbleto interface
in viscous starts 24 (8),
liquid, Langmuir,
4388 -4393
decreasing air column width
 Use and
application of
Microbubbles*
Optical micrograph of
monodisperse bubbles
100µ
m
produced in Lipid-water
suspension using the T-
junction*.
Ultrasonic transmission
response to bubbles
produced using different
techniques. A) Blue and grey
plot shows water and
suspension response. B) Red
dotted line shows T-junction
bubbles
*K.Pancholi, U.Farook, R.Moaleji, E.Stride,M.Edirisinghe (2008),Novelresponse*.
Method for preparing phospholipids coated
microbubbles, European Biophysics Journal, Volume 37, Number 4.
 • When ultrasound pulse is
transmitted through
microbubble suspension
backscatter from the
microbubbles only contain
one frequency
• Backscatter depends on
a b
microbubble radius, gas
core and microbubble
coatings
• Particles surrounding
increases stiffness of the
bubble in response to
ultrasound
• Results in response
containing(2008)
*E. Stride, K. Pancholi, M. J. Edirisinghe and S. Samarasinghe multiple
Increasing the
nonlinear character of microbubble oscillations atfrequency
low acoustic pressures, Journal of
Royal Society Interface, 5(24):807-11
 Ultrasound response of nano-
particle coated microbubbles
a b

• Experimental measurements of backscattered

ultrasound power
showing the amplification of nonlinear behaviour. The
black curve
shows the frequency spectrum for the backscattered
power from a
bubble suspension without nano-particle and the grey
 Physics of fluid
• Slender body theory applied to drop 75

• Drop placed in hyperbolic flow 70

•Tangential flow boundary condition

65

〈 ec〉
s1

60

•Normal stress balance 55

50
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

•Plotted radius of droplet against length

M

of droplet in direction of flow 75

• Both liquid viscosity (µl) and the ratio

of liquid to gas flow rate (Ql/Qg) 70

influences the size of the bubble 65

•with higher flow rates and higher s1

〈 ec〉

viscosities smaller bubbles form r 60

but only up to a limiting value of Ql/Qg
and/or µl. 55

dR g Po Rg
(z + G G )  9  50
−1 3 2
= − 1 − R 1 + G3G1−1 z 2 l 2 
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

3 1 zl M

dt 2µ G1  2 
Z
*K.Pancholi, E.Stride, M.Edirisinghe (2008) Dynamics of bubble in viscous liquid, Langmuir, 24 (8),
4388 -4393
 The angle between liquid supply and
Two liquid supply capillaries. The gas supply is shown 45 degrees.
angle between liquid supply and gas
supply could vary from 0-90 degrees

The angle between liquid supply and

gas supply is shown 0 degrees and
no third capillary. In all other cases
The angle between liquid supply and third capillary can be avoided as
gas supply is shown 0 degrees. well.

Possible designs of the T-junction

*UK patent application No. 02776963. PCT filed on 28/03/08 (PCT/GB2008/001050)
 Hybrid method*
Manometer

T- Harvard PHD- •Without electrospraying bubble

junction 4400 syringe
pump size of
Pressurized
Air Tank 30 mm with ± 0.1μm standard
Dimple
deviation.
Potential
V electrical •After electrospraying bubble size
Collection
vial
difference obtained
applied
was 5.1μm with ± 2μm standard
a b
deviation.

• Stability of bubbles found to be 2

hours
without any change in size.
20µm
Optical micrograph of microbubbles
collected directly from the outlet of the T- •Bubbles produced with this
junction a) without an applied electric method
field and b) with electric field. guarantees
*K.Pancholi, E.Stride, M.Edirisinghe (2008),Journal of targetedthat
drug all bubbles
delivery, are less
16(6):494-501.
 V

Insonation of chitosan suspension and subsequent electrospraying of foam in

sodium hydroxide solution.
•Increase in chitosan concentration in
solution controls the release of drug.

•Variation in acetic acid in chitosan

solution can reduce surface tension
without decreasing viscosity.

•Electrospray of highly viscous liquid

increases the particle size but lowering
surface tension results in smaller Chitosan particle size distribution for different
particles. surface tension but nearly same viscosity.

Ketan Pancholi, Eleanor Stride and Mohan Edirisinghe (2008), Novel preparation of porous
chitosan particles for drug delivery, Journal of Materials in Medicine (submitted)
 a b c

d e f

SEM images of chitosan particles made using electrohydrodynamic

atomisation of chitosan foam. Foam was prepared by insonation of
chitosan suspension. (a) Particle is of 500 nm range (b) Bulk particles
(c-d) one particle out of bulk (e-f) probable porous membrane or void
created by exposure of foam bubbles on surface.
 Image correlation analysis

4µm
Re-distribution of receptors after exposure to receptor specific antibodies and confocal
image of microbubbles

•Useful for measuring kinetics of particles at micrometer scale.

• Enables counting number of nano-particles present on image.
• Velocity, diffusion coefficient and direction of molecules can be measured.
•The method can be extended to other types of characterising techniques.
• Possible applications measurement of drug diffusion, interaction of signalling
molecules with receptors and intracellular transport of any vector.

1. Ketan Pancholi, Eleanor Stride and Mohan Edirisinghe (2008) Spatio-temporal image correlation spectroscopy for ultrasound
assisted drug diffusion measurement.
 Image correlation spectroscopy to measure
diffusion

•Experimental procedure •Simulation showing the temporal

•Real time image obtained using high fluorescent intensity fluctuations due to
speed camera diffusion of fluorescent dye from particle in
•Fluorescence intensity of images were water.
auto-correlated spatially and temporally1 •Darkest blue is lowest intensity and darkest
red represents highest intensity
1. Ketan Pancholi, Eleanor Stride and Mohan Edirisinghe (2008) Spatio-temporal image correlation spectroscopy for ultrasound
assisted drug diffusion measurement.
 Estimation of bubbles
diffusion
• Microbubbles
containing
fluorescent dye
immobilised in water
like gel
• Fluorescence
intensity of images
were auto-correlated
spatially and
temporally*
• Nonlinear data fitting
to function to find
out diffusion constant
*Ketan Pancholi, Eleanor Stride and Mohan Edirisinghe (2008) Spatio-temporal image
correlation spectroscopy for ultrasound assisted drug diffusion measurement,