Supplementary Information for

A microfluidic device for real-time on-demand intravenous oxygen

delivery.

Ashwin Kumar Vuthaa,b Ryan Patenaudea, Alexis Colea, Rajesh Kumara,b, John N. Kheira,b and
Brian D. Polizzotti a,b,c *
a
Department of Pediatrics, Harvard Medical School, Boston, MA USA
b
Department of Cardiology, Boston Children's Hospital, Boston, MA USA
c
Experimental Therapeutics Program, Dana Farber Cancer Institute/Harvard Cancer Center,
Boston, MA USA

*Brian D. Polizzotti

Email: brian.polizzotti@gmail.com

This PDF file includes:

Figures S1 to S12

References

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Figure S1 - Schematic of Generation 1 device

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Figure S2 - Schematic of Generation 2 device

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Figure S3 - Schematic of Generation 3 device

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Figure S4 - Schematic of Generation 4 device

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Figure S5. Schematic of optimized microfluidic device with all relevant
dimensions. Schematic illustration of the 3D multinozzled microfluidic device used
for in-situ nanospraying. The arrangement of sequential nozzles of decreasing
dimensions allowed generation of bubbles in the sub-micron regime. The 3D expansion
was introduced to alleviate the adverse pressure drop requirement for pumping a two-
phase mixture through a series of microscale nozzles. Such a step has previously been
shown to aid the formation of sub-micron particles (1)

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Fig. S6. Optical profilometry measurement of the SU-8 mold used to fabricate the
microfluidic devices. The device consisted of two layers, with the second layer in the form
of a step of ~ 25 µm height. (a) 3D view of the SU-8 mold and (b) Measurement of step
heights across the section indicated by the inclined red line.

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Figure S7. Representative images of the operation of the multinozzled device, as
observed in between the first and second nozzles. When the flow was far from steady-
state for a given ratio of flow rates (a), the gas phase formed a wide bulb-like shape with
frequent interfacial oscillations that lead to the formation of large bubbles at the outlet
of the device. At steady-state (b), the width of the gas stream reached a minimum and
the interfacial oscillations were greatly reduced or non-existent, resulting in the smallest
size distribution of bubbles that could be obtained for a given ratio of gas and liquid flow
rates. The figure presents a representative change in gas stream width with time. Actual
widths depended on the ratio of gas and liquid flow rates. Scale bar is 50 µm.

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Figure S8: Varying foam fractions seen at different gas and liquid flow rate with the
multinozzled microfluidic device. The coarse foam at the top of the sample indicated the
presence of larger bubbles formed due to coalescence at higher bubble concentrations,
which rise faster due to buoyancy forces. The images were captured a few minutes after
collection of the sample from the device. During in-vivo operation, the bubbles on exiting
the outlet tubing, would immediately come in contact with desaturated blood, thus aiding
their dissolution and reducing coalescence.

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Figure S9. Human donated blood was desaturated using 95% N2/5%CO2. Infusion of
oxygen microbubbles from microfluidic devices rapidly raised oxygen tensions as
evidenced by the color change from deep maroon (Left, desaturated blood) to bright
red (right, saturated blood).

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 Microbubbles
Free gas bubbles
Pale Lungs

Figure S10. Animals were infused with oxygen nano/microbubbles continuously for
30 min followed by a 30 min observation period. Microbubbles are clearly evident in
the RV (white patch, A). Free gas pockets are evident in the SVC and IVC (B). Lungs also
appeared pale suggesting significant microvascular obstruction at the end of the study
period (A). Qg = 0.7 sccm; Ql = 12mL/hr.

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 No microbubbles
in RV
Pink Lungs

Figure S11 . Animals were infused with oxygen nano/microbubbles continuously for
30 min followed by a 30 min observation period. Necropsy revealed no evidence of
bubble collection in the RV. Lungs also appeared healthy and pink with no evidence of
obstruction or hemorrhage at the end of the study period. Qg = 0.4sccm; Ql = 12mL/hr.

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Figure. S12: Experimental setup for size distribution analysis of the lipid micro/nanobubbles
produced using serial shear. Oxygen was supplied to the microdevice from a pressurized cylinder
(Airgas, Inc.). The liquid phase was pumped through the device using a high-precision syringe
pump (Harvard Bioscience, Inc). Bubbles produced from the device were diluted in a vial contained
clean DI water, prior to making size measurements. In the case of the in-vivo experiments, the
collection vial was eliminated and the outlet tubing from the device was inserted into the femoral
vein of a male Sprague-Dawley rat which was instrumented with an angiocatheter.

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Fig. S13. Photomicrograph of the actual nanospraying device. (Inset) optical
photomicrograph of the multi-nozzle geometry.

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 (Bubble concentration, #/mL)
2.5×105 **** 0.25 ml O2/min
2.0×105 ** **** 0.4 mL O2/min
**** 0.7 mL O2/min
5
AUC

1.5×10 **** Ql = 12 mL/hr

**
1.0×105 ****
****
5.0×10 4

0.0
1

5
5-

1-

2-
0.

Diameter (mm)

Fig. S14. Bubble concentration as a function of bubble diameter at varying gas flow
rates (Qg). Given the limitations in our experimental setup these values likely represent a
gross overestimation of the actual microbubble diameter exiting the device.

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References

1. S. A. Peyman, et al., Expanding 3D geometry for enhanced on-chip microbubble

production and single step formation of liposome modified microbubbles. Lab
Chip 12, 4544–4552 (2012).

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