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A Miniaturized Programmable Multi-Fluidic Pneumatic System for precise control of Sample Preparation Environment

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Abstract

High-density microfluidics is becoming an important experimental platform for studying complex biological systems such as synthetic gene regulatory networks, molecular biocomputating of engineered cells, distributing rapid point-of-care diagnosis, and monitoring pathological environment. Imaging transient bio-chemical reactions happening in these systems at a single particle or cellular level requires precise time-dependent control of sample reaction and imaging conditions at the desired fluidic momentum. In this study, we showed our novel miniaturized and programmable electronic-based pneumatic system to meet the requirement. We demonstrated its capability to control reaction parameters such as concentrations and injection rates in a liposome production system.
Lawrence Berkeley National Laboratory
Recent Work
Title
A MINIATURIZED PROGRAMMABLE MULTI-FLUIDIC PNEUMATIC SYSTEM FOR PRECISE
CONTROLS OF SAMPLE PREPARATION ENVIRONMENT
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https://escholarship.org/uc/item/1q37q9td
Authors
Narayanasamy, Sankar Raju
Vasireddi, Ramakrishna
Holman, Hoi Ying N
Publication Date
2021-10-14
License
https://creativecommons.org/licenses/by-sa/4.0/ 4.0
Peer reviewed
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University of California
A MINIATURIZED PROGRAMMABLE MULTI-FLUIDIC PNEUMATIC SYSTEM FOR
PRECISE CONTROLS OF SAMPLE PREPARATION ENVIRONMENT
Sankar Raju Narayanasamy1,2, Ramakrishna Vasireddi3 and Hoi-Ying Holman1,2
1. Berkeley Synchrotron Infrared Structural Biology Imaging Program, Lawrence Berkeley National Laboratory, Berkeley,
USA; 2.Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, USA;
3. Proxima-1 & Microfluidics Laboratory, Synchrotron SOLEIL, Saint-Aubin, France
ABSTRACT
High-density microfluidics is becoming an important experimental platform for studying complex biological
systems such as synthetic gene regulatory networks, molecular biocomputating of engineered cells, distributing
rapid point-of-care diagnosis, and monitoring pathological environment. Imaging transient bio-chemical reactions
happening in these systems at a single particle or cellular level requires precise time-dependent control of sample
reaction and imaging conditions at the desired fluidic momentum. In this study, we showed our novel miniaturized
and programmable electronic-based pneumatic system to meet the requirement. We demonstrated its capability to
control reaction parameters such as concentrations and injection rates in a liposome production system.
KEYWORDS: Microfluidics, miniaturized pneumatic system, sample preparation, sample introduction, liposomes
INTRODUCTION
The intricacy of biological processes can be understood by observing the transient bio-chemical reactions as
they happen using high-density microfluidic chips-based imaging studies1,2. Very large scale integration of
microfluidic channels such as those in high-density microfluidic chips enables researchers to study these reactions.
Existing commercially available sample injection methods are based on syringe pumps or pneumatic pressure
control pumps which employ lengthy tubing and can be costly. Presently, researchers at imaging user facilities use
large power-electrical based pneumatic systems4 to precisely introduce samples/materials into the integrated
microfluidic devices. As the field of biological and biomedical sciences advances, there is an emerging need for
more portable and affordable pneumatic systems to accurately introduce complex samples into high-density
microfluidic reactors while satisfying the compact space requirement. To meet this need, we develop a miniaturized
versatile electronic-based pneumatic system that can be easily adapted to project-specific sample preparation and
introduction. This portable system can be scaled up in multiples of 4 miniature pneumatic pumps. Furthermore, it
is only a fraction of the total cost of existing commercially available sample injection apparatus.
EXPERIMENTAL
Our palm-size multi-fluidic pneumatic sample injection device with high-precision sample handling is shown
in Figure 1. In the demonstrated liposome synthesis experiment utilizing this new miniaturized pneumatic pressure
control system, we used ambient filtered air as pressure feed for the diaphragm pumps. The air can be replaced with
specific suitable gas as per biological sample compatibility if needed. The flow rates of each sample introduced into
the channels is controlled independently.
INSTRUMENTATION & RESULTS
The miniature pneumatic pumps are connected to the compact multi-board, which in turn is coupled to the high-
drivers and Raspberry Wifi header to meet all inter-connectivity needs. Multiple multi-boards can be easily stacked
up in a custom built 3D printed holder. The droplet-stable giant unilamellar vesicles synthesizing microfluidic
device3 was fabricated using maskless lithography procedure, utilizing Dilase-250 table-top high-resolution laser
lithography system. We have demonstrated the utilization of the miniaturized programmable multi-throughput
multi-fluidic pneumatic system for the synthesis of cell-like liposomes such as droplet-stable giant unilamellar
vesicles (dsGUV) in the 10-100 µm range (Figure 2).
CONCLUSION
The high-precision multi-fluidic bio-sample preparation and injection system can be easily installed in any
bioimaging microscopic setups satisfying the compact space requirement in many imaging facilities. Thus this setup
is a crucial advance for complex synthetic biology studies. Furthermore, the setup can be extended to multi-level
sample injections in single particle and cellular cryo-electron microscopy(cryo-EM) sample preparation systems.
Figure 1. (a) An experimental setup of 2 single 4-pumps units. (b) Schematic representation of a single 4-pumps unit. This unit
can be easily scaled up as per the number of samples to be controlled and the flow rates for each pump is controlled via
PC/Laptop using multi-board application suite.
ACKNOWLEDGEMENTS
This work was conducted through the Berkeley Synchrotron Infrared Structural Biology (BSIS-B) Imaging
program, supported by DOE Office of Biological and Environmental Research, under contract no. DE-AC02-
05CH11231. The authors also would like to acknowledge the technical team of Bartels Mikrotechnik GmBH,
Germany for providing the certain components and specifications to establish this multi-fluidic bio-sample injection
setup.
REFERENCES
[1] T. Thorsen et al, Science, 298, 2002.
[2] A.J. van der Linden, J. Vis. Exp, 152, 2019.
[3] S. Deshpande and C. Dekker, Nat. Protocols, 13, 856-874, 2018.
[4] K. Brower et al, HardwareX, 3, 117-134, 2018.
CONTACT
* Hoi-Ying N. Holman; phone: +1-510-486-5943; hyholman@lbl.gov
Figure 2. (a) Octanol assisted
liposome (OLA) assembly
based microfluidic device fabri-
cated using maskless lithogra-
phy for synthesizing cell-scale
micro- liposomes. (b) Sche-
matic of the computer aided de-
sign of OLA microfluidic de-
vice. (c-d) Manually produced
dsGUVs in the range of 5-25
µm and 30-110 µm. (e) Micro-
fluidic produced 30-60 µm
dsGUVs, and (f)the envisioned
100-µm dsGUVs to be pro-
duced.
ResearchGate has not been able to resolve any citations for this publication.
  • T Thorsen
T. Thorsen et al, Science, 298, 2002.
  • A J Van Der Linden
A.J. van der Linden, J. Vis. Exp, 152, 2019.
  • S Deshpande
  • C Dekker
S. Deshpande and C. Dekker, Nat. Protocols, 13, 856-874, 2018.
  • K Brower
K. Brower et al, HardwareX, 3, 117-134, 2018.