Microfluidic Device for the
Detection of V. cholerae in
Drinking Water
Cholera Microbusters, Inc.
CEO: Dr. Orlin Velev
Designers: Jeffrey Millman, Katherine Schadel,
Chris Shaul, Lao Yang
Overview
Goal and Motivation
Cholera: Cause and Detection
Available Detection Tools
Silver Enhancement Technique
The Microfluidic Advantage
Device Concept
Optimization of Design Features
Future Challenges
Cholera Microbusters, Inc.
Goal
To design a small, accurate, and inexpensive
microfluidic device for the detection of
cholera
Help local health officials identify outbreaks
Provide residents and travelers with an affordable,
discrete, and quick means of assessing risk
Cholera Microbusters, Inc.
Motivation
Cholera
Fatal if not promptly
treated
Predominant in Asia,
Africa, Latin America
Mass infections in
refugee camps and
after natural disasters
IV drip treatment of cholera victims at
Problematic in tropical a refugee camp in Mozambique.
areas popular with [Medecins Sans Frontiers/Doctors Without Borders]
[Link]
tourists reatment/multimedia/msf_moz_cholera_camp.jpg
Cholera Microbusters, Inc.
Motivation: Scope of the Problem
[Link]
Cholera Microbusters, Inc.
Cholera: Cause and Detection
Vibrio cholerae
Aquatic bacterium
Cholera toxin (CT)
Chlorine ion
imbalance
Severe dehydration
Resistant to Vibrio cholerae
vaccination and [Link]
antibiotics
Most devices detect CT
only
Cholera Microbusters, Inc.
Available Detection Tools
Naturally Specific Antibodies
Labeling
Fluorescent markers
Gold colloids
Immunoassay with Gold-Colloid Labeling
Courtesy Shalini Gupta, NCSU chemical engineering graduate student
Cholera Microbusters, Inc.
Commercially Available Device
The SMART
Gold-Labeled Antibodies
Limit of detection 6 million cells
per mL
Portable
External Requirements
Buffer
Reaction vial
Eyedroppers The SMART by New Horizons
Diagnostic Corporation (Columbia
Swabs
Maryland, USA)
[Link]
Cholera Microbusters, Inc.
Silver Enhancement Technique
Amplified level of
detection
Implemented on
microfluidic devices
recently
Immunoassay with Gold Labeling No known
and Silver-Enhancement
Courtesy Shalini Gupta, NCSU chemical engineering environmental or safety
graduate student
hazards
Cholera Microbusters, Inc.
The Microfluidic Advantage
Microchannel
Prototype
Smaller devices
RNA
Drastically reduced Fingerprinting
reagent volumes
Decreased waste DNA
Assays
Examples of previously developed
microfluidic devices currently on the
market.
Courtesy Dr. Orlin Velev, NCSU chemical engineering professor
Cholera Microbusters, Inc.
Device Concept
Credit-card-sized Assay and
diagnostic device result
with reagent and Sample
reaction chambers collection
Cholera Microbusters, Inc.
Device Considerations
Polydimethylsiloxane
(PDMS) Rubber
Elastomer
Antibody binding
techniques studied
Ease of microfabrication
Finger-Actuated Micro-
Pump The importance of rounded
chambers. (Ahn et al. 2004)
Sample collector
Reagent dispenser
Cholera Microbusters, Inc.
Finger-Actuated Micro-Pump
Optimization
Purpose
Chamber Wall Determine effective
Chamber volume
Deliver sufficient volume
Constraints on
Diameter
Large enough to be
easily handled
Small enough to fit on a
Finger-Actuated Micro-Pump microfluidic device
Cholera Microbusters, Inc.
Theoretical Analysis
Chamber Wall Chamber Wall
Dead Volume Dead Volume
Effective Volume Effective Volume
Chamber with Elastic Top Chamber with Elastic Top and
and Bottom Rigid Bottom
⎡ 2 ⎛ R 2 + rR + r 2 ⎞⎤
Vd = π h ⎢ R − ⎜⎜ ⎟⎟⎥
⎣ ⎝ 3 ⎠⎦
Cholera Microbusters, Inc.
Theoretical Analysis
80
60
% Dead Volume
40
20
0
0.5 0.6 0.7 0.8 0.9 1 1.1 1.2
Diameter (inches)
Percent dead volume decreases as diameter
increases
Not a function of height
Cholera Microbusters, Inc.
Experimental Analysis
Two Scenarios
Sample collector
Reagent dispenser
Constructed from Silicone
Rubber
Independent Variables
Diameter
Thickness
All numbers
in millimeters
Cholera Microbusters, Inc.
Reagent Dispenser Experimental Data
Height
80.0%
0.0625 inches
0.0313 inches
60.0% 0.0200 inches
% Dead Volume
40.0%
20.0%
0.0%
0.3 0.5 0.7 0.9 1.1
Diameter (inches)
Percent dead volume decreases as diameter
increases
Not a clear function of height
Cholera Microbusters, Inc.
Sample Collector Experimental Data
80.0%
60.0%
% Dead Volume
40.0%
Height
20.0% 0.0625 inches
0.0313 inches
0.0200 inches
0.0%
0.3 0.5 0.7 0.9 1.1
Diameter (inches)
Percent dead volume increases as diameter
increases
Not a clear function of height
Cholera Microbusters, Inc.
Chamber Optimization Results
Sample Collector
1-inch diameter
Larger size because of higher percent dead volume
Reagent Dispenser
0.625-inch diameter
Smaller size because of lower percent dead volume
Smaller amount of reagents
Height
0.0625-inch
No effect on dead volume
Greater total volume
Cholera Microbusters, Inc.
Channel Sizing Optimization
Guiding Principles
Low fill-up time of about 1 second
Manufacturing costs
Device size
Method
Extrapolation from experimental data
Mechanical energy balance with estimates of frictional
terms (length, expansions, contractions, bends)
Cholera Microbusters, Inc.
Channel Sizing Optimization
6
5
Fill-up Time (s)
0
0.4 0.6 0.8 1 1.2 1.4 1.6
Channel Width (mm)
Optimal channel width of 1 mm
Cholera Microbusters, Inc.
Five-Pump Design
All numbers
in millimeters
Cholera Microbusters, Inc.
Five-Pump Design
Cholera Microbusters, Inc.
Five-Pump Design
Cholera Microbusters, Inc.
Conclusions
Finger-actuated
No external
requirements
Portable
Disposable
Smaller, simpler
alternative to
commercially
available options
Cholera Microbusters, Inc.
Future Challenges
Final device selection
Build prototype
Operating procedure
Valve system to prevent backflow
Economic analysis
Cholera Microbusters, Inc.
Acknowledgements
Dr. Orlin Velev’s
research group, for
their help and
access to laboratory
equipment
Dr. Lisa Bullard, for
her advice and
[Link]
assistance ghostbusters/[Link]
Cholera Microbusters, Inc.