An Introduction to Microfluidics Thursday, October 12th, 2017

Michael De Biasio 1003446167

1. Introduction
Microfluidics – the study of fluids on a microscopic scale – has important applications in
biological research and industry, such as in DNA sequencing and microchip applications [1]. In
this experiment, micro-fluid flow situations are analyzed and observed using fluorescent beads in
fluid, a microscope and an image capture software. The lab’s purpose is to observe, investigate,
and gain insight into the properties of fluid flow through microscopic channels.
Theoretical Background
Fluid flow can either be laminar – smooth with no path interference and low velocities –
or turbulent –high velocities, characterized by whirlpools and turbulences [2]. Furthermore, all
fluids have viscosity due to friction, and are constrained by the no slip condition – that the fluid
velocity be zero at a rigid surface [3]. Thus, a fluid velocity profile has both viscous and inviscid
flow regions, where the frictional effects are either significant and insignificant, respectively.
By Conservation of Mass, a steady, incompressible flow has an inlet flow rate equal to
the outlet flow rate, expressed in the continuity equation:
𝑚̇ = 𝜌𝐴1 𝑣1 = 𝜌𝐴2 𝑣2 (1)
where 𝑚̇ is the mass flow rate, 𝜌 is the density, 𝐴 is the cross-sectional area, and 𝑣 is the velocity
[4]. This analysis of a constant density, steady fluid flow can be extended through Bernoulli’s
equation:
1 1
𝑃1 + 𝜌𝑣12 + 𝜌𝑔ℎ1 = 𝑃2 + 𝜌𝑣22 + 𝜌𝑔ℎ2 (2)
2 2
where P is the pressure, 𝜌 is the density, 𝑣 is the velocity, 𝑔 is gravitational acceleration, and ℎ is
the elevation [5]. Thus, if you know the relative speeds and heights of two points in a fluid flow
analysis, the pressure difference between those points can be calculated.
2. Experimental Procedure and Results
The equipment and procedure from the laboratory manual [1] was used.
A. Hemacytometer Scale
To measure the distances travelled by the beads in the fluid flow, the hemacytometer was
used as a scale, with Figure 1 from [1] used for reference distances in millimeters. The photo-
editing tool “Pixelmator” (like Photoshop) was used to measure distances via pixel lengths.
Results are shown below in Table 1, along with a picture of the hemacytometer in Fig. 1.

Table 1: Hemacytometer Measurements at 10x Magnification

Total Picture Length 1.25 ±0.05 mm 1392 ±2 pixels
Total Picture Height 0.95 ±0.05 mm 1040 ±2 pixels
Sample Square 0.05 ±0.001 mm 55 ±2 pixels
Conversion factor k 0.00091 ± 0.00004 mm/pixel

Note: 10x magnification and an exposure rate of 671.0±0.05 ms

was used for all data. Fig. 1: 10x magnification
of hemacytometer
 B. Straight Channels

a) b)
Fig. 2: (a) Straight channel path, with streaks representing moving beads. (b) Velocity Profile
for the straight channel path
To calculate the velocity of the fluid, the streak lengths were measured, and using the
exposure rate, the velocity at different points along the channel were calculated via:
𝑑
𝑣= (3)
𝑡
By calculating the velocity at different points across the channel, a velocity profile for the path
can be formulated and is shown in Fig. 2(b). The velocities in the profile range from 0.038mm/s
near the wall to 0.079 mm/s in the middle of the channel, and some representative velocities are
shown below in Table 2. In addition, a sample calculation for velocity is shown below. Note that
at the wall, by the no-slip condition assumption, the tangential velocity of the fluid is zero.
𝑚𝑚 (562 − 534) ± 2 𝑝𝑖𝑥𝑒𝑙
𝑣 = (0.00091 ± 0.00004 ) = 0.038 ± 0.003 𝑚𝑚/𝑠
𝑝𝑖𝑥𝑒𝑙 0.671 ± 0.00005 𝑠
Table 2: Some Streak Velocities for the Straight Channel
Distance from Bottom Velocity (mm/s) Distance from Bottom Velocity (mm/s)
of Channel (mm) of Channel (mm)
0.052 ± 0.003 0.047 ± 0.003 0.31 ± 0.01 0.079 ± 0.004
0.19 ± 0.009 0.079 ± 0.004 0.39 ± 0.02 0.071 ± 0.004
0.28 ± 0.01 0.079 ± 0.004 0.52 ± 0.02 0.038 ± 0.003
C. Channels of Different Sizes

a) (b)
Fig. 3: (a) Gradual change in channel width. (b) Abrupt change in channel width.
 The streak velocity was calculated using equation (3) both before and after entering the
channel for the gradual and immediate width change. These speeds were taken as streaks from
the middle of the channel to ensure no changes due to frictional forces from interaction with the
channel walls. The values for these calculations are shown in Table 3 below.
Table 3: Data for Velocities from Gradual and Abrupt Width Changes
Type of Entrance Average Entrance Exit Width Average Exit Calculated Exit
Change Width (mm) Velocity (mm/s) (mm) Velocity (mm/s) Velocity 𝐴1𝑣1 (mm/s)
𝐴 2
Gradual 0.38±0.02 0.073±0.004 0.16±0.01 0.13±0.01 0.18±0.02
Abrupt 0.55±0.02 0.103±0.005 0.18±0.01 0.20±0.02 0.30±0.03

As seen in Table 3, Equation (1) can be used to compare the two quantities of 𝐴1 𝑣1 and
𝐴2 𝑣2, by calculating the expected final velocity and comparing it to the experimental value. This
comparison shows that the mass flow rates are not proportional within error, as would be
expected by the continuity equation. In both cases, the experimental and calculated values differ
by approximately 30%.
D. Bent Channels

a) b)
Fig. 4: (a) Sharp turn in bend. (b) Curved bend, more gradual turn.
Once again, the bead velocities were calculated from both images in Fig. 4, both before
and after the bend. The results are shown in Table 4 below. For the two bends, results verified
that the velocity does not change as the beads pass through the bend.
Table 4: Data for Different Bend Scenarios
Bend Initial Velocity (mm/s) Final Velocity (mm/s) Difference (mm/s)
Sharp 0.52±0.02 0.49±0.02 0.03±0.03
Curved 0.052±0.004 0.056±0.004 0.004±0.005

3. Error Analysis
Measurement Error
To determine the measurement uncertainty, a few estimations and assumptions had to be
made about the error of obtained data. Firstly, an error of ±2 pixels was used in image analysis,
as the photo editing software used, Pixelmator, gives precise information about pixel length.
Thus, the only error in estimating the streak distances was from determining the beginning and
end of the streak, with a maximal variance of 1 pixel on each end. In addition, since no error on
the hemacytometer was listed, it was assumed that the given box width of 0.05mm had an error
range of 0.001mm. Finally, the exposure rate had an assumed error of 0.05ms, as the software’s
accuracy is to the first decimal digit.
From these assumptions, all error was propagated using the following error propagation
relationships [6]:
𝑞 =𝑥 ±𝑦 ±𝑧 ⇒ 𝛿𝑞 = √(𝛿𝑥)2 + (𝛿𝑦)2 + (𝛿𝑧)2 (4)

𝑥𝑦 𝛿𝑥 2 𝛿𝑦 2 𝛿𝑧 2
𝑞= ⇒ 𝛿𝑞 = 𝑞 √( ) + ( ) + ( ) (5)
𝑧 𝑥 𝑦 𝑧
For example, propagating the error from the sample calculation in section 2.B:
0.038𝑚𝑚 0.00004𝑚𝑚/𝑝𝑖𝑥𝑒𝑙 2 2𝑝𝑖𝑥𝑒𝑙𝑠 2 0.00005𝑠 2
𝛿𝑣 = ( ) √( ) + ( ) + ( ) = 0.003𝑚𝑚/𝑠
𝑠 0.00091𝑚𝑚/𝑝𝑖𝑥𝑒𝑙 28𝑝𝑖𝑥𝑒𝑙𝑠 0.671𝑠

Systematic Error
The main source of error in the lab arose from equipment malfunctioning. When
measuring the fluid flow through the channel, our apparatus never had a steady flow rate, with
the fluid flow often stopping and moving in the opposite direction of the expected flow direction.
Furthermore, air bubbles often got stuck in the tubing, which led to no fluid flow through the
channel. In this case, the inlet fluid syringe needed to be pressurized to push the air bubble
through the channel and allow for fluid flow to resume. Overall, the general inconsistency in
fluid flow made it more difficult to capture images of the flow.
Additional equipment error resulted from the clumping of beads along the channel. This
is visualized well in Fig. 2(a), where the large yellow dots represent clumps of beads. These
stationary beads create barriers to the laminar fluid flow, forcing fluid to flow around them.
Thus, they represent an imperfection in the channel and a source of error. For example, the
curved, bent channel shown in Fig. 4(b) has a large number of stationary beads. This makes it
hard to discern fluid flow streaks, effecting the ability to accurately discern inlet and outlet
velocities of the curved path.
In the future, both of these sources equipment error could be accounted for by either
using new channels, or by finding a way to clear stationary beads from the channels so that
undisturbed fluid flow can occur. While these errors cannot be quantified without further
investigation into the channel apparatus used, they have a large effect on the results obtained in
the experiment, and must always be considered when analyzing quantitative data.
Finally, the channel itself is not straight as predicted. The unevenness of the channel is
represented visually in Fig. 5 (in Fig. 5(a), the large vertical imperfection is assumed to be below
the channel, most likely a piece of dust/dirt caught underneath). As can be seen at both 20x and
40x magnification, the channel walls have minor bumps which can impede velocity flow. By the
no-slip condition, the velocity of the fluid is zero at the wall due to frictional effects. Since the
wall is not perfectly planar as a result of imperfections, these frictional effects are not uniform
across the surface. However, because the imperfections are minor when compared to the overall
channel size, they seem to have no impact on the velocities near the wall in data collection.
Therefore, their effect can be viewed as negligible when analyzing channel flow.

(a) (b)
Fig. 5: Wall Imperfections at (a) 20x and (b) 40x Magnification
4. Discussion
B. Straight Channels
The maximum velocity of the fluid is in the center of the channel, as outlined in the
velocity profile shown in Fig. 2(b). This is expected, as it is the region in which the fluid is
farthest from the channel walls, and thus will experience the least viscous effects. Furthermore,
due to this maximum velocity and the no-slip condition on the channel wall, the greatest velocity
difference is expected to occur between the center and edges of the channel.
The observed flow is laminar, as it produces smooth, straight velocity streaks with no
apparent whirlpools or turbulences. This velocity can be manipulated in a number of different
ways. First-off, the velocity would increase by lifting the syringe with the inlet fluid to a higher
height, as the fluid falls through a greater distance under the same gravitational acceleration. In
other words, the fluid’s potential energy is converted into kinetic energy of fluid flow. Secondly,
by forcing the fluid through the micro-channels via pressurization, the velocity of the fluid
through the channel can be greatly increased (this technique was often used to expel air bubbles,
as described in Section 3 above). Both of these examples for increasing the velocity of fluid flow
can be seen as direct applications of Bernoulli’s principle (equation (2)), assuming the flow is
constant and steady, as changes in height/pressure at the inlet are effecting channel velocities.
C. Channels of Different Sizes
As the fluid flow passes from large to small channel widths, the velocity increases. In this
transition from slow to fast velocity, the laminar flow curls inwards to fit in the width of the
smaller channel. As depicted in Fig. 3, this flow curl for abrupt width change is much more
pronounced and sudden, while gradual width change has a continuous transition between widths,
and thus a broader inward curl of fluid velocities.
Table 3 used equation (1) to calculate the predicted exit velocity. In both gradual and
abrupt width changes, calculated exit velocity was higher than the experimental velocity, with a
percent difference of 28% and 33% for the gradual and abrupt width changes, respectively. This
large margin of error can be attributed to the incorrect assumption of steady fluid flow made via
use of the Continuity Equation (equation (1)). As discussed in Section 3, the fluid flow did not
exhibit steady flow, as the rate of mass transfer into and out of the system was changing. Thus,
the assumption that flow could be represented by the Continuity Equation was imprecise.
 D. Bent Channels
As represented by the difference column in Table 4, the velocity before and after the
Fig.6:
bends did not change to a significant amount within the given error range. This is expected by
Path
Bernoulli’s equation (equation (2)) as pressure, potential energy and fluid density are constant. Lines for
Laminar flow in both cases was observed. As discussed in Section 3, the curved bend channel Curved
had many stationary beads, making flow observation harder than the sharp bend channel. Bent
However, the fluid’s path was apparent in the sharp bend channel as depicted in Fig. 4(a), and Channel
therefore this type of flow, where the fluid follows through the channel bend in a laminar flow
fashion, can be extrapolated for the gradually curved bent channel, as shown in Fig. 6.
Holistic Observations
Comparing all the channels, the inaccuracy of a constant fluid flow as discussed in
Section 3 makes any velocity comparison impossible. In addition, if the fluid had no pressure
(i.e., no force pushing it through the channel via gravity or pressurization), then the fluid would
remain stationary in the channel and not flow. Finally, in general, these results could be
reproduced with much more accuracy if better micro-channels were used.
5. Conclusions
This experiment analyzed the flow of fluid in various micro-channels, and compared
experimental results with theoretically expected results. In general, the data obtained was in
accordance with the theoretical background. The result with the most deviance from the expected
value was the velocity difference for changing channel widths. This deviance was attributed to
lack of steady flow as assumed in the Continuity Equation. However, all channels exhibited
laminar flow, and it was shown, within uncertainty, that the velocity across bent channels does
not change, in accordance with Bernoulli’s principle. In addition, a velocity profile adequately
represented the viscous effects and no-slip condition for fluid flow. For future experiments,
higher quality results can be obtained if new channels with fewer stationary beads are used.
6. References
[1] Keith, B., Chung, E., & Dell, T., Intro to Microfluidics, 3rd ed. A. Shukalyuk & B. Mark,
Eds. Toronto: University of Toronto, 2013.
[2] “What is the difference between laminar and turbulent flow?”, Physlink.com. [Online].
Available: http://www.physlink.com/education/askexperts/ae464.cfm.
[3] Abidin, U., “Chapter 1: Introduction”, Fluid Mechanics I. University Putra Malaysia
[Online]. Available: http://www.fkm.utm.my/~ummi/SME1313/Chapter%201.pdf.
[4] “Continuity Equation”, Princeton.edu, 2017. [Online]. Available:
https://www.princeton.edu/~asmits/Bicycle_web/continuity.html.
[5] “Bernoulli's Equation”, Princeton.edu, 2017. [Online]. Available:
https://www.princeton.edu/~asmits/Bicycle_web/Bernoulli.html.
[6] "Propagation of Error", Chemistry LibreTexts, 2017. [Online]. Available:
https://chem.libretexts.org/Core/Analytical_Chemistry/Quantifying_Nature/Significant_Digits/Pr
opagation_of_Error.