Senior

Design Spring 2015

Final Report
By: Samuel Beck, Bradley Bernard, Rheagan Chambers,
Kristen Galloway, Brandon Landry, Alexandra Williams

Faculty Advisors: Noelle Bryan, Dr. Brent Christner, Dr. Steven Hall, Charles Malveaux

1 | P a g e

Table of Contents:
Abstract 5
Introduction .. 6
Problem Statement .... 7
Measurable Objectives .... 7
House of Quality ... 7

Figure 1 .. 8
Product Design Specification 9
Design Alternatives .. 10

Function Structure Diagram . 10

Figure 2 10

Figure 3 11
Physical Decomposition .. 11
Figure 4 .. 12

Morphological Chart .. 13
Table 1 ... 14
FALL 2014 MATERIAL .. 15

Preliminary Designs ... 16
Summary of Construction to Date . 17

Figure 5 . 17

Materials of Construction .. 17

Table 2 .. 18
Graph 1 .20
Table 3 . 21
Figure 6 .. 21
Graph 2 . 22
Graph 3 . 22

Control System ..... 23

Figure 7 . 28

Efficiency Factors .. 28

Figure 8 . 29

Graph 4 . 29
Figure 9 30
Table 4 .31
Table 5 . 31
Figure 10 31
Figure 11 31
Figure 12 32
Calculations 33
Final Design ... 35
Figure 13 35
Methods of Analysis ... 36
Future Schedule .. 38

2 | P a g e


Figure 14 38
Budget ... 39

Table 6 39
Bill of Materials ... 40

Table 7 40
Conclusion .. 41
SPRING 2015 MATERIAL 42
Final Design 43

Figure 15 43

Figure 16 44
Materials .... 44
Construction Methods 44
Figure 17 46

Figure 18 46
Control System 46

List of parts .. 49

Table 8 49
Wiring Schematic ... 50

Figure 19 50
Actual Circuit Board... 51

Figure 20 51
Testing ... 51

Figure 21 52

Graph 5 .... 52
Graph 6 .. 53
Graph 7 .. 53
Fluids Dynamics 54
Figure 22 54
Graph 8 .. 55
Graph 9 .. 56
Bacterial Collection ... 56

Figure 23 58

Figure 24 .. 58
Figure 25 58
Figure 26 58
Redesign ..... 58
Schedule ..... 59
Figure 27 59
Budget ..... 59
Table 9 60
Bill of Materials ...... 61
Table 10 61
Conclusion ..... 62
References .. 63
Appendix 1: Decision Matrices .... 64

Table 11 .. 64

3 | P a g e


Table 12 .... 64

Table 13. .. 64

Table 14 .. 64
Appendix 2: Impact ..... 65

Figure 28 65
Figure 29 65
Figure 30 66
Appendix 3: Impingement .. 67
Figure 31 67
Figure 32 67
Appendix 4: Impact + Impingement ... 68
Figure 33 68
Appendix 5: Inducted Fan .. 69
Figure 34 69
Appendix 6: Materials .. 70
Figure 35 70
Figure 36 71
Figure 37 72
Figure 38 73
Graph 10 74
Appendix 7: Fluid Dynamics ... 75
Table 15 .75
Appendix 8: UAS-Hexacopter ...... 76
Table 16 76
Graph 11 . 76
Graph 12 . 77
Table 17 77
Table 18 78
Figure 39 . 79
Table 19 80
Appendix 9: List of FAA Regulations ............ 81
Appendix 10: Control System ... 82
Figure 40 82
Figure 41 83
Figure 42 84
Figure 43 85
Figure 44 86
Figure 45 87
Figure 46 88
Appendix 11: Graphical Analysis .... 89
Graph 13 89
Graph 14 89
Appendix 12: Calculations ... 90

4 | P a g e

Abstract:
The ACROBE project consists of a remotely operated bacterial collection device
designed for the purpose of furthering aerial bacteria data and aiding in the relatively new
field of bio-precipitation. Three current methods exist for capturing bio-aerosols, but these
methods do not allow for the control of variables such as collecting at known altitudes. The
ACROBE offers high repeatability and reproducibility as well as ease of use for potential
users. The ACROBE will rely on the use of an inducted fan that creates the devices vacuum
pressure. Two identical impactors will allow for increased sampling volume per flight.
Servo operated doors ensure the device remains sealed until collection at target altitudes. A
collection plate will hold industrial grade aluminum foil coated with a film layer of glycerol,
which will reduce the bounce effect of the bacteria. Various sensors will be on board for
data collection of weather conditions. To ensure optimal positioning of the device, a
lightweight wind vane can be used in combination with a rotary encoder. The device will be
predominately constructed of carbon fiber due to its low density. The entire system will be
mounted on an Unmanned Aerial System or more specifically a Hexacopter.

5 | P a g e


Introduction:

Bio-precipitation is a fairly new field. It is the study or concept of particular

microbes that highly organize the water molecules so that the freezing temperature of the
water is increased by 7 C [1]. These microbes are called ice-nucleating (IN) bacteria, and
they are known for being plant pathogens. In the United States, nearly one billion dollars go
to waste potentially due to these ice-nucleating bacteria causing frost damage to the crops
[2].
The three current methods for studying bacteria in the atmosphere include the
HASP (High Altitude Student Platform), hailstones and ice core samples. The purpose of the
HASP is to carry twelve student payloads to approximately 36km high with the duration of
each flight lasting between 15-20 hours [3]. The HASP, however, does not allow the
determination of which altitude the collection of bacteria occurred. Hailstones are studied
because they have ice-nucleating bacteria at their cores. The problem with this method is,
in order to collect these IN bacteria, one has to wait for hail and then wait more time for the
hailstone to melt. Time is a key issue for studying bacteria through the use of hailstones.
Last, the ice core samples are taken from the North Pole. This method of collection is
inconvenient due to the location being far. Overall, the three current methods allow for low
variable control.
In order to study IN bacteria, a device for microbe collection will be built to allow
scientists to learn more about the bacteria and where it is located in the atmosphere. This
device is called the ACROBE.

6 | P a g e

Problem Statement:

The ACROBE is designed to develop an efficient device that will collect microbes in

order to increase bacterial data at known altitudes satisfying Federal Aviation

Administration (FAA) regulations. This will be accomplished by mounting the ACROBE to
an Unmanned Aerial System-Hexacopter (UAS-H).

Measurable Objectives:

There are a total of four measurable objectives for ACROBE. After discussing with

Dr. Brent Christner, a microbiologist here at Louisiana State University who recently had an
article published in Nature concerning bio-precipitation, the device should filter at least
1 ! of air to ensure a large enough sample size. Since the ACROBE is designed to
determine at which elevation the bacteria exists, the ACROBE should collect within 2 of
a specified altitude. Finally, the hexacopter has the ability to carry up to 15 , so the
ACROBE will weigh less than 10 .

House of Quality:

For the House of Quality, a scale of 1, 3 and 9 was used to rank the correlation

between engineering characteristics and customer requirements with 1 being the least
correlated and 9 being the most correlated. The customer requirements were listed on the
vertical axis and engineering characteristics on the horizontal axis. Next, each
corresponding box was given a ranking. After tabulating the results, remote operation of
the device was found to be the most important engineering characteristic.

7 | P a g e


Figure 1: House of Quality for the ACROBE.

8 | P a g e


Product Design Specification:

The construction materials for the device will mainly be carbon fiber. In order to

make the ACROBE easy to repair and disassemble, the design will be modular. The device
will also be designed for reproducibility and reliability to ensure that others who use this
will be able to produce the same outcomes. Based on preliminary testing where a pressure
sensor was placed in a tube while the fan ran for roughly a minute at approximately
!

20 ! , the operation vacuum pressure was found to be near 0.03 ! or 206.84 ! . Since
the device will be flown in Louisiana, over crop fields, operation temperatures will be
between 40 F and 100 F (4.44 C and 37.78 C) due to that being the normal, average
range of temperatures year round. The sampling time will range from 2.5 to 20 .
The minimum of 2.5 is the amount of time it takes to collect exactly 1 3 while 14
minutes is the maximum amount of time the UAS-H can stay in flight. For cleaning
purposes, 70% ethanol and 2000 ppm of NaClO will be used. Sampling will be done in
compliance with FAA regulations, which state that the UAS-H cannot be flown over 400
from the ground and no closer than 5 to an airport.

9 | P a g e


Design Alternatives:
Function Structure Diagram

The function structure diagram directly below shows that bacteria, electrical energy,

a control system and air flow will enter the ACROBE in order to achieve bacterial collection.
Collected bacteria, mechanical energy, a control system and air flow will be outputs of the
device.

Figure 2: The above picture is a simplified representation of the Function Structure
Diagram for the device.

Electrical energy drives the control system. In the more complex function structure
diagram, Figure 3, it is shown that the ACROBE is driven by a pressure differential. A
negative feedback exists where this pressure differential controls the speed of the DC
motor.

10 | P a g e

Figure 3: This is a more complex Function Structure Diagram for the collection device.

Physical decomposition

The ACROBE can be divided into two main components: the impactor, of which

there will be two, and the electronics. From there, each of these two components are
further decomposed into more subunits. Refer to Figure 4 for a more detailed description.

11 | P a g e


Figure 4: Physical decomposition for the ACROBE.

12 | P a g e

Morphological Chart

The morphological chart allowed us to determine the best solutions for each

component of the design before the detailed design stage began. For bacterial collection,
impact was ranked the best, followed by impingement. Originally, both impact and
impingement were the two chosen methods of bacterial collection. In a conversation with
Dr. Christner, he expressed his thoughts on impingement. His opinion along with not
finding many papers that praised impingement, the decision to completely remove
impingement from the device and to only have two impactors was made. For the platform
that would be used to carry the device, a UAS-H outnumbered the rest of the ideas due to
stability during flight and cost. In order to be able to control the device from afar, remote
and automated data logging was the best solution. Next, for bacterial extraction, agitation
of the foil allows the glycerol, which contains bacteria to dissolve into a solution of choice.
For the differential pressure source, an inducted fan was chosen. Last, for cleaning
purposes, 70% ethanol and NaClO are the best solutions for the device. The strength of
70% ethanol, in particular, was chosen because any percentage higher than 70 would dry
out the cell membrane rather than killing them. Having at least 30% of water allows the
organism to remain hydrated enough so that the solution can penetrate the cell. Once the
solution reaches the inside of the cell, it will denature the proteins and dissolve the lipid
bilayer [4]. Ethanol will be used to clean the outer surface of the device, while the NaClO
will be used to bathe the components of the device that come into direct contact with the
bacteria.

13 | P a g e


Table 1: This morphological chart was created based on our decision matrices, which can
be found in the Appendix 1: Decision Matrices.

14 | P a g e

Fall 2014 Material

15 | P a g e

Preliminary Designs

Our original design included both an impact and impingement system. The impact

was the first method of bacterial collection. A funnel containing a grid of small rods would
have been coated with a viscous fluid to capture the bacteria. After further thought, the
impact system was changed to having a coned funnel with a 120 angle that would cause
the air to flow through the inlet and come into contact with a Trypticase Soy Agar (TSA)
plate. From there, a hood was created that contained two tubes to allow the air to flow into
the impinger after the impaction stage. The purpose of this change was based on Marple
and Willekes method [5]. This method is used to ensure maximum collection efficiency.
The pressure differential caused unforeseen problems with the impingement. Large
bubbles were created which could potentially lead to loss of bacteria. To help this problem,
we created a grate with holes with 0.0625 diameter to reduce bubble size. The
impinger was changed from a cube to a cylinder. At the same time in the design process, the
inlet tube was moved from the side of the impinger to the bottom. The geometry and
position changes were made to lower the amount of liquid needed. Even though a grate
was made, it did not solve the bubble size and bacteria loss problem to the extent that it
was needed. An inverted cone was placed in the impinger to help control the creation of
bubbles, their location and their size. Original designs can be found in Appendix 2: Impact,
Appendix 3: Impingement and Appendix 4: Impact + Impingement.

16 | P a g e

Summary of Construction to Date:

Figure 5: Exploded view of impactor design.

Materials of Construction
Selection of materials for the ACROBE were chosen in order to meet physical
construction and biological criteria. Materials were chosen based on primary and
secondary factors. Primary factors include low density, high corrosion resistance, non-
cytotoxicity, and low cost. Secondary factors, such as smooth surface texture properites,

17 | P a g e

high electrical resistivity, and high strength, were considered only to components of
interest that were constrained to unique design features.
Shown below is a table of physical construction materials and the pertinant
characteristics of each:

Table 2: Physical construction materials.

The majority of physical construction material was chosen to be carbon fiber, since
it meets all specified primary and secondary criteria. Though carbon fiber is slighly more
dense and more expensive than nylon, it is still preferred since carbon fiber is widely
available in a variety of woven fabric forms (uni,bi, tri-directional) and fiber thicknesses
(tow). Woven fabrics are advantageous in prototyping, such as in ACOBE construction,
because it allows for increased design flexibility in material mechanical properties and
geometry. In order to permenately set woven fabrics into solid shapes, epoxy resin is
required [6]. For this reason, epoxy will also be a major physical construction material.
West Systems 105/206 epoxy was chosen since it meets all primary criteria and is user
friendly [7].
Supplementary physical materials are required because not all of the ACROBEs
physical sub-components can be carbon fiber. To satisfy the need for airtight inlets and an
outlet door, neoprene gasket material will be used. This material is easily deformed, which

18 | P a g e

allows it to fill any void spaces between two mating faces. Aluminum 6061-T6 may be
required if electrostatic force via induction become problematic. More specifically, the tube
brackets and 37 tubes of 0.1 diameter, shown in Figure 5, will be aluminum in order to
electrically ground them to the Arduino common ground. 6061-T6 is a specific alloy of
aluminum that can be easily machined and is affordable due to its abundance.
Unfortunately, 6061-T6 will oxidize to aluminum oxide (Al2O3) when treated with sodium
hypochlorite (NaClO). This constraint can be overcome by either applying a surface coating
(anodize) or using marine grade aluminum, which better resists corrosive conditions [8].
Nylon 6,6 is a thermoplastic, synthetic polymer that will be incorporated into the
impactor design in order to increase modularity. The three 1.6 inch nylon bolts and six
hand grip style nuts shown above allow for complete disassembly of the hood and
collection disk from the collector base. Other material options for bolts and hand grip style
nuts come with a better strength (secondary factor) but at the cost of excess weight
(primary factor). For this reason, nylon bolt and nut assemblies were chosen [9].
In general, construction materials were down selected with non-cytotoxicity as first
priority followed by low density. Since the ACROBE is being attached to a hexacopter
platform, the weight of the ACROBE must be minimized to allow for longer collection times.
In order to illustrate the dependence of weight in achieving maximum flight times, the
graph below was generated:

19 | P a g e


Graph 1: Overall flight time of the UAS-H (RCTx800 + ACROBE) is dependent on the weight of
the ACROBE itself, which is currently unknown. Assumptions made in calculations include
80% battery capacity, constant current drain (hovering flight only), and constant battery cell
voltage. An expected ACROBE weight of 600g yields a max flight time of 15 minutes.

Three major biological materials were selected to collect, culture, and inhibit the
growth of collected bio-aerosols. The ability to encourage or disallow cell growth before,
during and after collection was a chief concern in biological material selection because
there is an inherent time delay between bio-aerosol collection and analysis. A reduction in
possible change in sample size, due to post-collection growth, is desired in order to
maintain representative aero-environment data. Collected bacteria will be preserved on
the ACROBEs collection plate via a film layer of glycerol, which has been shown in
preliminary experiments to not allow for cell growth, yet sustain cell viability. Glycerol is a
polar compound, making it miscible in other aqueous polar solutions, such as PBS and the
final biological material- R2A. Phosphate Buffered Saline (PBS) is also able to maintain cell
viability without allowing cell reproduction; however, PBS is much less viscous and will
continue to flow at even film thicknesses. If quantification tests are being conducted, PBS
will be used as the solvent for the bacteria. Reasoners 2A is a nutrient broth that selects

20 | P a g e

for and cultures slow growing bacteria, like the prominent bio-aerosol Pseudomonas
syringae.
Substrate
(mg/L)
203.3
148.0
80.0
63.3

Time
6
8
11
13

1/S
(L/mg)
0.0049
0.0068
0.0125
0.0158

(1/hr)
0.1335
0.1054
0.0800
0.0690

max
(1/hr)
7.4889
8.4092
9.4912
10.4921

1/
(hr)

Km
(mg/L)

av
(1/hr)

td (hr)

0.1545

38.96

0.097

7.148

Table 3: Biological Kinetics Data of P. syringae in R2A

In the scenario that collected bacteria will be cultured in order to produce a larger
sample size, the quantification of growth rate, cell concentration vs. absorbance, doubling
time, half-saturation coefficient, and substrate utilization of P. syringae is desired. To
determine this, the following experimental process was conducted, which yielded the
relevant biological kinetics:

Figure 6: Experimental design for quantification of P. syringae growth, cell concentration,

and substrate utilization in R2A.

21 | P a g e

Lineweaver-Burk Plot of P. syringae in R2A

12
y = 252.16x + 6.4706
R = 0.99759

10
1/ (hr.)

8
6
4
2
0
-0.03

-0.02

-0.01

0
1/[S] (L/mg)

0.01

0.02

Graph 2. Lineweaver-Burk plot, showing the biological kinetics data of P. syringae in

R2A.

Cell Concentrations (cells/mL)

Cell Concentrations of P. syringae During Log Growth Phase

5.5E+09
y = 4E+08x
R = 0.95805

5E+09
4.5E+09
4E+09
3.5E+09
3E+09
2.5E+09
2E+09
5

9
10
Time (hr.)

11

12

13

14

Graph 3: Cell concentration data obtained by correlating absorbance data with cell
plating results.

22 | P a g e

Control Systems

The microbial collection device will optimally be used 400 in the atmosphere

during collection. Therefore, the device needs to be controlled remotely and must have a
receiver on board to communicate with a transmitter. Furthermore, the device needs to
remain sealed until sampling begins. Because of this, the two entrance and one exit doors
will need to be servo operated. A control system will allow the servos to open and close
once the device reaches the correct height. The device will operate at multiple
temperatures, varying from 40 F to 100 F. Weight will be one of the most important
factors since the device will be attached to a hexacopter. All equipment must be as light as
possible. The purpose of the device is to allow scientists to study IN bacteria effectively.
Consequently, the device will need to be outfitted with a range of sensors for weather
conditions and data collecting capability. One goal of the ACROBE is to have high collection
efficiency. Moderately high winds could potentially ruin efficient collection if the device is
not facing the wind. Therefore, the device should sense which direction the wind is
blowing and allow the intake to face that direction.
The on-board fan that creates the vacuum pressure will also need to be controlled
remotely so it can turn on and off during testing. Furthermore, the speed of the fan needs
to be monitored and controlled for continuous efficient collection. Cost is a key factor, so
the cost of the controls must remain low. The batteries must last at least 20 to
allow for flight time to and from the collection sight.

23 | P a g e

All sensors must be easily detachable as well because the device will be modular and be
cleaned frequently. In short, the constraints can be listed and simplified:
o Cost
o Weight
o Temperature
o Sterile
o Efficient Collection
o Battery Life
o Ease of Use
o Data Collection
o Remotely Controlled

Arduino boards are built for a specific project, but the Arduino Uno and Arduino Mega
can be used for many different types of projects. Between these, the Uno does not have as
much computing power or I/O pins. However, the Uno is cheaper and lighter. After finding
all the sensors needed for the collection device, it was apparent that they could not all fit
and be used on the Uno. The Arduino Mega would need to be used.
The Arduino Mega was chosen as the microcontroller for the microbial collection
device. The Uno is based on the Atmega2560 microcontroller. It has 16 analog input pins
and 54 digital I/O pins. The digital I/O pins contain 15 pins capable of PWM. This board
provides enough space for all the sensors needed. It has a clock speed of 16 MHz and a
RAM of 8 . It also operates on 5V with a DC current requirement of 40 mA on all I/O
pins. The hardware on board can talk Serial (UART), I2C and SPI. Outside the chip, the
Arduino Mega board contains voltage regulators, capacitive components, and I/O
connectors. The code written in the integrated development environment (IDE) is the only
code that runs on the chip. There is no interpreter, operating system, or firmware. On the
other hand, the Raspberry Pi is a single board computer, or SBC. On board it contains a 32
bit microprocessor and supports video, audio, USB host, ethernet, SD card, HDMI cord, and

24 | P a g e

GPI headers. These assets mean the Raspberry Pi has more in common with a computer
than an Arduino. Instead of writing code to control the hardware directly, the programs
written are actually run within an operating system. The operating system is typically
Linux. Both the Arduino and Raspberry Pi contain a CPU, timers, memory, as well as I/O
pins. The key difference is in the I/O pins. As a microcontroller, the Arduino has high I/O
capability, able to drive external hardware directly. On the other hand, the Raspberry Pi is
a microprocessor, which tend to have weak I/O, needing transistors to drive most
hardware. However, microprocessors are effective processors containing better brain
power than a microcontroller. The Raspberry Pi has a 700 MH clock speed, over 40 times
faster than the Arduino. It is also based on a 32-bit architecture, compared to the 8-bit of
the Arduino. The Raspberry pi has 256,000 times more RAM than an Arduino. The
Arduino has more I/O pins and can also utilize 8 times more current in each pin than the
Raspberry Pi. Finally, the Raspberry Pi typically operates on Linux, while the Arduino has
no operating system. The Arduino was ultimately chosen for ACROBE for its capability of
controlling many sensors with more I/O capability, and, its simple, effective interface, while
remaining cost friendly.

Modern wind direction sensors on the market are heavy, expensive, and not

designed to interface with the Arduino. The final design selected for wind sensing was the
rotary encoder. A rotary encoder will be placed in between the wind vane and intake.
Specifically, the EMS22P50-B28-LS6 from Bourns. It has low friction and high sensitivity.
The rotary encoder will determine the relative position of the intake to the vane. A negative
feedback loop will be created between the rotary encoder and a continuous rotation servo
attached to the intake. When the vane is not lined up with the servo, the servo will turn.

25 | P a g e

Once lined up, the servo will stop. A potentiometer could have been used to determine the
position of the wind vane, however, it could increase friction on the wind vane. The more
friction the wind vane has, the less sensitive it will be to wind speed. The hexacopters
position also needs to be known, and a potentiometer cannot be used for this. A digital
compass sensor could have been used, but many of them would have to be used to
orientate the position of the vane, intake, and hexacopter.

The speed of the fan drawing air through the device is imperative for efficient

collection of bacteria. The speed of the air moving through the inlets must be in between
!

20-30 ! . The final design will have a pressure sensor, specifically the MPX4115a, located in
a pitot tube near the end of the system to measure the pressure when the air speed in the
!

inlets is 20-30 ! . The pressure value taken will then be used in a negative feedback loop to
drive the fan. If the pressure ever reaches over or under a normal value, the fan will either
reduce or increase its speed. The fan will be controlled by an S-12 ESC. The ESC speaks to
the Arduino using servo language, and thus can be controlled using PPM from the servo
library. The ESC is lightweight and contains the power requirements to operate the DC
motor. The pressure sensor is a rugged, temperature compensated and cost effective
sensor that functions within the specified pressure and temperature range.

Humidity is important for studying bio-precipitation. Temperature is also good data

to have when studying cells that live in ambient weather. Humidity is related to
temperature change, so many breakout boards will read humidity as well as temperature.
The HIH 6130 breakout board from Sparkfun was chosen. It operates from 2.3 to 5.5 volts
and is temperature compensated from 5 to 50 C. It also has a compensated humidity
range of 10% to 90% relative humidity. The HIH 6130 speaks to the Arduino through the

26 | P a g e

I2C bus. The HIH 6130 will be placed in a protective location from debris but will remain
open to the environment for data acquisition.

The solution for data acquisition needed to be lightweight and cost effective. Most

DAQs are extraordinarily expensive, heavy and consume copious amounts of space.
Adafruit manufactures an SD card shield that outmatches all of the criteria. Data can be
sent to the lightweight and portable SD card, ready for acquisition upon placing the card
into a computer. The shield is placed on top of the Arduino, saving space and weight.

Two batteries will be used for the entire system. An 11.1V, 3S Lipo battery will

power a 12A ESC and the DC motor used to drive the fan that causes suction. The other
batteries will be two 14.4V, 4S Lipo battery. This battery will power the six 20A ESC used to
control the hexacopters rotors. In addition, the 5V BEC onboard the ESC will run to the RX
receiver. The receiver will then turn on the Arduino when commanded from the
transmitter.

27 | P a g e

Wiring Schematic:

Figure 7: Schematic of control system. Microcontroller featured in the center. DC motor is

powered by 11.1V Lipo battery. Power source is continuous 5 volts from receiver.

Efficiency Factors
The fluid flow throughout the device was analyzed using Autodesk Simulation CFD.
The main goals of fluid analysis were to visualize fluid flow and to obtain velocities and
Reynolds numbers throughout the device.

A cross sectional plane through the middle of the device is shown below in Figure 8.

It is color coordinated depicting low, medium and high velocities, respectively blue, green
and red. In addition, the picture confirms that air impacts with the plate before going

28 | P a g e

around the plate and exiting through the outlet. A velocity curve through the middle of one
of the tubes is also shown in Graph 4. The graph shows that our simulation produced
maximum velocities at the center of the tube. The maximum velocity is approximately
!"

1095 ! (27.8 ! ).


















Figure 8: A cross sectional photo displaying velocity flow throughout the device.

















Graph 4: A graph of the velocity at the middle of one of the small inlet tubes.

29 | P a g e

Figure 9 below displays a cross-section of the 37 small inlet tubes in the design. The

design is symmetrical across both the x and y axes, giving rise to tubes that are spatially
equivalent, which are color coordinated. Table 4 shows the average Reynolds numbers and
velocities of each set of equivalent tubes. Autodesk Simulation CFD provided the flow rate
through each small tube and using = , the average velocity per tube was calculated.
The main trend within this data is that velocity and Reynolds number increase slightly
moving toward the center. The percent standard deviations were calculated for each set of
tubes for both velocity and Reynolds number. The minimum and maximum values are
displayed in Table 4. These values show that the standard deviations are fairly small.

















Figure 9: Cross sectional photo of the 37 inlet tubes, which are color coated to relate
spatially equivalent tubes.

30 | P a g e














Table 4: Average Reynolds numbers and velocities of the spatially equivalent tubes.

Table 5: Range of percent standard deviations for Reynolds numbers and velocities for
spatially equivalent tubes.

Particle trace animations can be performed using Autodesk Simulation CFD. A

picture of this animation is below and to the left. On the right, velocity vector field is shown.

Figure 10: A particle trace showing the

airflow throughout the collector.

Figure 11: The velocity vector field throughout

ACROBE.

31 | P a g e

After observing the velocity vector field, airflow behaves as predicted, except for the
whirlwind that occurs in the corners of the device in front of the collection plate. Possible
redesign to reduce this could be to chamfer those edges or bring the collector plate closer
to the 37 inlet tubes.

For efficient bacterial collection, certain requirements are needed. First, the S/W

ratio should be between 1 and 5. The S value is the distance between the collection plate
and the exit of the inlet tubes. The W is the width of each inlet tube. Second, the throat
length must be greater than or equal to the nozzle diameter with nozzle diameter also
being W. Third, the Reynolds number must be between 500 and 3000. Fourth, the velocity
!

of the air through the inlet tubes must be 20 ! to 30 ! .

Figure 12: Streamlines and particle trajectories for a typical impactor.

32 | P a g e

Calculations:
50%collection diameter (Yao and Mainelis, 2007)
!" =

9/! ! ! * !"

= .

The d50 equation was used to calculate the d50 specific to the ACROBE. At 2 m, the
device should collect 50% of all bacteria with this diameter. 2 m is a little more than
average size of P. syringae; however, we can theoretically collect up to 16 times the
minimum 1 m3 of air volume. For this reason, we can justify collecting less than 50% of P.
syringae. If any bacteria are collected with a larger diameter than 2 m, more than 50% of
bacteria collected.

Diameter of small inlet tubes (max Re=3000)

=

= . .

Volumetric Flow Rate
=

= .

33 | P a g e

Given a Reynolds number range of 500 to 3000 based on Marple and Willekes
method, we calculated the smallest diameter for the inlet tubes. Using 3000 as the Reynolds
!

number and 20 ! for velocity in the above equations, we calculated that the minimum
diameter must be approximately 0.1 inch. Using this diameter, the volumetric flow rate, per
!

tube, was calculated to be 1x10-4. !

Number of tubes needed in order to filter 1 of air in 10 minutes

= 1.0134

!!
!

!"!
!!"#

!"!"#
!

= 0.0608! 10

0.0608!
= 1!
1
=

Diameter of outlet pipe necessary to equal of 37 tubes
!"! = = 37 5.067110!! ! = 1.87510!! !
!"! = 1.87510!! ! =
= .

!"# !
4

.
= .

Using the volumetric flow rate, 0.06078 m3 of air would filter through per tube, per
ten minutes. In order to filter the wanted 1 m3, it was calculated that 17 tubes were needed.
By doubling the number of tubes to 34, we can increase the amount of air filtered through

34 | P a g e

the device. We chose to have 37 tubes in each impactor in order to create symmetry about
the x and y axes.

Final Design:
The final design includes two impactors that will be placed on top of the hexacopter.
The hexacopter will also hold the Arduino system, a wind vain and a control. In Figure 13,
you can see the ACROBE mounted on top of the hexacopter. On the sides of the hexacopter
are two covered compartments; one housing a control and the other housing the
electronics for the device.

Figure 13: The ACROBE mounted onto the UAS-H.

35 | P a g e

A detailed, exploded view of an impactor was shown earlier in Figure 5. As

previously stated, the ACROBE sill have servo-operated doors sealed when the device is not
collecting. During collection at a specified altitude, the door will be open. The bacteria will
enter through a 120 angle funnel and continue along and travel through 37 tubes,
measuring 1 length and 0.1 diameter each. The dimensions of these tubes allow
for fluid flow dynamics that will achieve the optimal efficiency (based on simulations ran in
Autodesk Simulation CFD). Next, the bacteria will adhere to glycerol that is coated on
industrial grade aluminum foil, which is placed on the collection plate. The glycerol is
viscous and reduces the bounce effect of the bacteria. From there, the air will travel around
the collection plate and exit the outlet tube that is .75 diameter. The entire impactor is
about 4 in length.

Methods of Analysis:
Following bacterial collection using the ACROBE, lab analysis is required to
determine total concentration and species of the bacteria, as well as, the viability status. To
prepare the bacteria captured in glycerol for lab analysis, a transfer to either Phosphate
Buffered Saline (PBS) or Reasoners 2A Agar (R2A) is required. PBS is a balanced salt
solution used often in biological research. The main function is to maintain pH and osmotic
balance, as well as, provide cells with water and inorganic ions. PBS can be used to
maintain cells for the short term in a viable condition while the cells are manipulated
outside the regular growth environment [10]. R2A is a low nutrient medium. When R2A
use is combined with lower incubation temperature and longer incubation time, it
stimulates the growth of stressed bacteria. Nutritionally rich media support the growth of
fast growing bacteria, and can suppress slow growing or stressed bacteria [11].

36 | P a g e

Analysis of bacteria can potentially be achieved in a number of ways and comprise a

number of methods. Plate counts are widely used for enumerating viable bacteria. Relying
on bacteria growing a colony on a nutrient medium allows this [12]. This method often
requires more time than others and can be inaccurate. The membrane filter method is
similar to plate counts. Bacterial cells are deposited on the upper surface of the membrane
filter are able to grow individual colonies when the filter is placed on suitable nutrient
surfaces like agar [13]. This method has an advantage in that large sample volumes can be
processed even with relatively low numbers of microorganisms. Another method is the use
of a hemocytometer, a device used to count cells, both alive and dead. A hemocytometer
can determine the bacterial concentration of a known volume. Bacterial viability can be
determined through a variety of stains such as the Gram Stain and the acid-fast stain.
Mass spectrometry has become a powerful and usable tool when it comes to
identification of bacteria. This method provides reliable information of bacteria
characterization even at a subspecies level [14]. A popular and all-inclusive molecular
method is Polymerase Chain Reaction (PCR). This method allows for the quantification and
identification of viable but non-culturable cells that are metabolically active but non-
dividing. All the previously stated methods can be used following bacterial collection with
the ACROBE depending on scientific purpose and preference.

37 | P a g e

Future Schedule:

Figure 14: The Gantt chart contains deadlines for both Fall 2014 and Spring 2015
semesters.

By creating Gantt chart, the progress of this project has remained on track. Next

semester, the ACROBE project will continue moving forward in order to build and test
while redesigning as needed. By the beginning of February, we would like to have all
control system parts working individually to ensure that all components function properly
before construction of the final design begins. Also, all other materials should have arrived
by this time. The beginning of April is when the final prototype should be finalized and
built. If time permits, bacterial collection and analysis will be performed as additions to the
goal of the project.

38 | P a g e

Budget:

Material
Propellor

Items
APC 13x10
APC 13x10

Adhesive

Epoxy
PVC
1-inx 5ft Sch-40 PI
3/4 in x 5 ft Sch40 PVC

2 in x 2ft PVC SCh 40 SOL

2 in sch 40 cap socket

1/2 in x 2 ft PVC Sch 40

1/2 in Sch40 cap

1 in sch40 elbow (3)

1 in sch40 coupling(2)

3/4 in sch40 elbow (3)

3/4 in sch40 adapter

3/4 in sch40 adapter

1 in sch40 adapter

1 in x 3/4 in adapter

8x10 clear lexan (3)
Unit
Fan
ss12 brushless 12 amp esc
lipo 2s 7.4V 1200 30c

ammo 20-40 3500KV brshless mtr

hyperflow 370 ep df w/o motor
Polycarbonate Polycarbonate tubing 1/4 ID
Polycarbonate tubing 3/4 ID

Polycarbonate tubing 7/8 ID

Polycarbonate tubing 1" ID

Polycarbonate tubing 1/2" ID

polycarbonate tubing 3/4 ID

actron vacuum and fuel pressure
Vacuum
kit
Sensors
Temperature and Humidty Sensor
ABS Pressure Sensor (x3)



Total

Price
$7.99
$7.99
$15.99
$3.09
$1.79
$4.14
$2.08
$3.03
$0.97
$1.98
$0.92
$1.41
$1.86
$0.47
$0.77
$1.56
$12.72
$34.99
$16.99
$42.99
$21.99
$20.25
$33.65
$16.11
$13.67
$25.09
$29.14
$17.84
$29.95
$48.27
$423.68

Table 6: The budget containing costs from both Spring and Fall of 2014.

39 | P a g e

The above table shows that the total cost thus far is $423.86, but due to already

having some of the materials, we have only spent $286.75 of the total budget of $1,100. At
the end of this current, fall semester, about $800 is still available for use in the spring.

Bill of Materials:
Item
MPX 4115a
HH6130 Sparkfun
Arduino Mega
EMS22P50-B28LS6
SD Card Shield Adafruit
Futaba ASD Sensor
Futaba BLS153
7.4 V 2S LiPo 1200 mAh
Futaba R7008SB S.Bus2 2.4 GHz FASSTest
Rx
CF Rod 1" x 60"
CF Rod .686" x 72"
CF Rod .1" x 24"
West Systems 105/206 Epoxy
West Systems 423 Graphite Powder
CF Plate 300mm x 100mm x 2mm
Frekote 700
Neoprene with Adhesive 54" x 1' x .125"
6061-T6 Aluminum 4" x 4" x .125"
Du-Bro 993 Nylon Bolt 3" x .25"
Aluminum Foil 18" x 25' x .001"
Teknova R2A Broth Premix (500g)
Growcells MRGF-6235-12 10X PBS (12L)
2000 ppm NaHClO3 (1L)
SEOH R2A Agar Media (500g)
20 pk. Petri Dishes 100mm x 13mm
Carbon Fiber Fabric 3K 50" x 36"
Heathrow Scientific HS86655 Stainless
Steel Large Bacterial Cell Spreader
Total

Quantity
3
1
1
1
1
1
3
1

Combined Price
$48.27
$29.95
$37.52
$37.80
$19.95
$179.97
$360.00
$16.99

1
1
1
2
1
1
2
1
1
2
1
1
1
1
1
1
10
1

$139.98
$75.99
$55.99
$4.90
$62.61
$19.03
39.99
$20.50
$20.00
$23.66
$2.36
$16.95
$176.57
$292.78

$14.76
$724.28

$75.52
$79.50
$50.85

Table 7: Bill of Materials for Spring 2015 Semester.

40 | P a g e

The Bill of Materials above includes all materials needed to complete the ACROBE.
The non-bolded items are the materials that are needed but that we do not need to
purchase do to already having them in privately owned possession. The bolded items are
the materials that still need to be purchased. We will spend a total of $724.28 in the spring,
remaining a under the $1,100 budget. However, for someone who plans to build the
ACROBE with no previously owned materials should expect to spend about $2,000.

Conclusion:

To end, the ACROBE is an ergonomic and easy to use design. It will allow for efficient

bacterial collection with high repeatability as well as reproducibility. Multiple runs will be
allowed in a day. Since we are operating at high altitudes, the device will be fully automated
allowing control of the ACROBE from a distance. Being fully automated, the device is
essentially a fire and forget system. The control system will allow for data collection with
the sensors placed within the device. Because the ACROBE will be able to collect at
specified altitudes, it is the first of its kind.

41 | P a g e

Spring 2015 Material

42 | P a g e

Final Design:

This semesters design has three major and impactful changes from the previous

design. The first change made was having one continuous door for both intakes, rather than
one door per intake. This change allows for one servo to be needed instead of two separate
servos to control the opening and closing of the doors. The next change we made was
adding flanges to seal the collectors simultaneously with ease. Last, the collection disk is
coated with silicone spray instead of glycerol. This change was made due to the glycerol not
remaining on the collection plate without sliding off and forming beads.

Figure 15: Exploded view of our final design.

43 | P a g e










Figure 16: Picture of our final design for the ACROBE.

Materials:

The physical and biological materials have not changed from last semester. Please

refer to page 17 for a complete list of the materials using for the ACROBE.

Construction Methods:
A variety of modern construction methods were used in both the prototype design
and the revised, final design. These methods include 3D printing, laser cutting, and water
jet CNC. In prototype construction, the main goal was to produce of viable model, which
could prove the mechanical concepts of the ACROBE, such as the rotation and fluid flow
requirements. Additionally, the prototype was needed in order to affirm the hexacopters
ability to fly with an elevated load and center of mass. Because the prototype was not
required to withstand chemical treatment, the main construction methods used were laser

44 | P a g e

cutting and 3D printing. The materials for these methods were plywood and ABS plastic,
respectively. Polycarbonate tubing was also heavily featured in the prototype design as a
pipe flow medium.
Transitioning to the final design, it saved both money and time to utilize some
prototype parts and pieces as a mold for final parts. The weight of the prototype ACROBE
was approximately 1.11 kg (2.45 lbs), which is an acceptable weight considering our design
objective of 10 lbs. By using molds and a lighter composite carbon fiber material, the
weight of the ACROBE was reduced 25% to 0.83 kg (1.83 lbs). Water jet CNC machining
was used to precision-make aluminum and carbon fiber plate parts. The CNC machine
enabled us to elevate both the quality of construction and expanded design options to more
complex geometries that would otherwise be extremely difficult to accomplish with
traditional methods.
Anodizing the aluminum parts was necessary for long-term successful operation of the
ACROBE, since 2,000 ppm bleach will oxidize bare aluminum. Basic steps for anodizing go
as follows:
1.
2.
3.
4.
5.
6.
7.
8.
9.

Remove all surface imperfections from aluminum.

Clean aluminum with soapy water.
Attach aluminum rod to part. This is the anode.
Arrange anodizing acid bath (sulfuric acid).
Place aluminum cathode in the bath.
Complete electrical circuit with 12V 8A DC supply.
Wait 20 minutes.
Remove part from bath. Submerge in dye (2 min).
Boil part in water for 1 min.

45 | P a g e









Figure 17: The above figure is a
picture of our prototype.

Figure 18: This figure is another angle of

the prototype.

Control System:
The Arduino Mega was chosen as the microcontroller for the microbial collection device.
The Mega is based on the Atmega2560 microcontroller. It has 16 analog input pins, and 54
digital I/O pins. The digital I/O pins contain 15 pins capable of PWM. This board provides
enough space for all the sensors needed. It has a clock speed of 16 MHz, and a RAM of 8 kb. It
also operates on 5V with a DC current requirement of 40 mA on all I/O pins. The hardware on
board can talk Serial (UART), I2C, and SPI. Outside the chip the Arduino Mega board contains
voltage regulators, capacitive components, and I/O connectors. The code written in the IDE is
the only code that runs on the chip. There is no interpreter, operating system, or firmware. On
the other hand, the Raspberry Pi is a single board computer, or SBC. On board it contains a 32
bit microprocessor and supports video, audio, USB host, ethernet, SD card, HDMI cord, and GPI
headers. These assets mean the Raspberry Pi has more in common with a computer than an

46 | P a g e

Arduino. Instead of writing code to control the hardware directly, the programs written are
actually run within an operating system. The operating system is typically Linux. Both the
Arduino and Raspberry Pi contain a CPU, timers, memory, as well as I/O pins. The key
difference is in the I/O pins. As a microcontroller, the Arduino has high I/O capability, able to
drive external hardware directly. On the other hand, the Raspberry Pi is a microprocessor, which
tend to have weak I/O, needing transistors to drive most hardware. However, microprocessors
are good at processing, containing better brain power than a microcontroller. The Raspberry Pi
has a 700 MH clock speed, over 40 times faster than the Arduino. It is also based on a 32-bit
architecture, compared to the 8-bit of the Arduino. The Raspberry pi has 256,000 times more
RAM than an Arduino. The Arduino has more I/O pins and can also utilize 8 times more current
in each pin than the Raspberry Pi. Finally, the Raspberry Pi typically operates on Linux, while
the Arduino has no operating system. The Arduino was ultimately chosen for ACROBE for its
capability of controlling many sensors with more I/O capability, and, its simple, effective
interface, while remaining cost friendly.
Modern wind direction sensors on the market are heavy, expensive, and not designed to
interface with the Arduino. The final design selected for wind sensing was the rotary encoder.
A rotary encoder will be placed in between the wind vane and intake. Specifically, the
EMS22P50-B28-LS6 from Bourns. It has low friction and high sensitivity. The rotary encoder
will determine the relative position of the intake to the vane. A negative feedback loop will be
created between the rotary encoder and a continuous rotation servo attached to the intake. When
the vane is not lined up with the servo, the servo will turn. Once lined up, the servo will stop. A
potentiometer could have been used to determine the position of the wind vane, however, it
could increase friction on the wind vane. The more friction the wind vane has, the less sensitive

47 | P a g e

it will be to wind speed. The hexacopters position also needs to be known, and a potentiometer
cannot be used for this. A digital compass sensor could have been used, however, many of them
would have to be used to orientate the position of the vane, intake, and hexacopter. A camera
may have to be used if the information from the digital compass cannot be sent remotely. A
camera sending live feed is not optimal because of the cost and added weight.
The speed of the fan drawing air through the device is imperative for efficient collection
of bacteria. The speed of the air moving through the inlets must be in between 20-30 m/s. The
final design will have a pressure sensor, specifically the MPX4115a, located in a port near the
end of the system to measure the pressure when the air speed in the inlets is 20-30 m/s. The
pressure value taken will then be used in a negative feedback loop to drive the fan. If the
pressure ever reaches over or under a normal value, the fan will either reduce or increase its
speed. The fan will be controlled by a S-12 ESC. The ESC speaks to the Arduino using servo
language, and thus can be controlled using PPM from the servo library. The ESC is lightweight
and contains the power requirements to operate the DC motor. The pressure sensor is a rugged,
temperature compensated, and cost effective sensor that functions within the specified pressure
and temperature range.
Humidity is important for studying bioprecipitation. Temperature is also good data to
have when studying cells that live in ambient weather. Humidity has to be temperature
compensated, so many breakout boards will read out humidity as well as temperature. The HIH
6130 breakout board from Sparkfun was chosen. It operates from 2.3 to 5.5 volts, and is
temperature compensated from 5 to 50 C. It also has a compensated humidity range of 10 to
90% relative humidity. The HIH 6130 speaks to the Arduino through the I2C bus. The HIH

48 | P a g e

6130 will be placed in a protective location from debris, but will remain open to the environment
for data acquisition.
The solution for data acquisition needed to be lightweight and cheap. Most DAQs are
extraordinarily expensive, heavy, and consume copious amounts of space. Sparkfun
manufactures a SD card data logger that outmatches all of the criteria. Data can be sent to the
lightweight and portable SD card, ready for acquisition upon placing the card into a computer.
The breakout board is placed on top of the Arduino, saving space and weight.
Two batteries will be used for the entire system. A 2S Lipo battery will power a 12A
ESC and the DC motor used to drive the fan that causes suction. The other batteries will be two
14.4V, 4S Lipo battery. This battery will power the six 20A ESC used to control the hexacopters
rotors. As well as this, the 5V BEC onboard the ESC will run to the RX receiver. Also, another
5V BEC will run from a separate ESC to power the CR servo. The receiver will then turn on the
Arduino when commanded from the transmitter.

List of Parts
Part

# of
Units

MPX4115a

HIH6130

Company
Manufacturer
Freescale
Semiconductor
Honeywell

Company of
Purchase
Digikey

Combined
Price
48.27

Amazon

29.95

Arduino Mega

Arduino

Amazon

37.52

EMS22P50-B28LS6
OpenLog

Bourns

Mouser

37.8

Sparkfun

Sparkfun

19.95

Futaba S148

Futaba

Adafruit

14

Futaba BLS153

Futaba

ServoCity

360

Total

547.49

Actual Price

187.49

Table 8: The total price is everything needed for construction of the control system. The
actual price only includes the prices of equipment that we had to buy.

49 | P a g e

Wiring Schematic

Figure 19: Schematic of control system. Microcontroller featured in the center. DC motor
is powered by 11.1V Lipo battery. Power source is continuous 9 volts from the switch.

50 | P a g e

Actual Circuit Board

Figure 20: Physical image of completed circuit board, showing data logger, temperature
and humidity sensor, voltage regulator, low-pass filter, and quick disconnects for the ESC
and CR servo.
Testing
The wind vane needed to be tested for its mechanical ability to face the wind direction.
The inherent ability for the wind vane to point towards wind direction was tested by placing the
wind vane onto the rotary encoder, and placing into a wind tunnel. In order to have efficient
collection, the wind vane needed to point within 15 degrees of wind direction.

51 | P a g e

Figure 21: Demonstrates the wind vanes direction from wind at three different wind
speeds. Red shows the angles the wind vane needed to stay within. Green shows the wind
vane when starting from 90 degrees to the left, and purple shows the wind vane when from
starting 90 degrees to the right.

The MPX pressure sensor was connected to the exhaust through a port. It is used to
measure the differential pressure caused by airflow. When the airflow speeds up through the
exhaust, the air becomes turbulent. The turbulent flow causes the pressure sensor to read errant
values. When these values are sent to the ESC, the motors speed fluctuates. A running average
was created in the code to smooth out the values sent to the ESC.

Graph 5: Demonstrates the wide standard deviation bars caused by no smoothing with
high throttle.

52 | P a g e
















Graph 6: Demonstrates the reduced error bars created by using a bin average.

Graph 7: Demonstrates significantly reduced error bars when a moving average is used.

53 | P a g e

Fluid Dynamics:
The ACROBE was analyzed using Autodesk Simulation CFD. When the small inlet
!

tubes had a velocity of 20 ! , the outlet velocity was found to be approximately 10 ! . Using
this value for the outlet velocity, an anemometer was used to monitor outlet velocity on the
!

ACROBE, and the inducted fan motor increased until the velocity was 10 ! . The internal
pressure at this time was recorded and used to calibrate the pressure sensor that drives
!

the inducted fan. This ensured that the velocity in the inlet tubes was the desired 20 ! .

Figure 22: Fluid flow analysis of ACROBE.

54 | P a g e















Graph 8: The velocity profile through one of the small inlet tubes of ACROBE, which shows
that the average velocity throughout most of the tube is approximately 21 m/s.


In order to determine where to place ACROBE relative to the hexacopter, we attached an
anemometer at various heights above the hexacopter, as wind speeds below the hexacopter
were much greater than above. The graph shown is an average of 7 data points per bar,
each at different angular positions on the same concentric circle. The y axis is the vertical
wind velocity and the x axis is the vertical distance above the main deck cover. It was found
that 4 above the main deck cover had the least amount of prop wash. This is where
ACROBE was mounted.

55 | P a g e

Graph 9: Graph showing the vertical wind speed with respect to distance above the main
prop deck.

Bacterial Collection:
Following bacterial collection with the ACROBE, the aluminum foil on which the
bacteria were collected was removed from the ACROBE carefully. To remove the foil, gloves
were worn and the foil was touched only on the edges to avoid removing any bacteria. The
ACROBE allows for two impact collections and both were used for separate purposes. The
first foil is stamped onto an R2A plate to create a press plate of the original bacterial
concentration. The second foil was transferred into 40 of R2A solution and shaken for
two hours to transfer the bacteria from the foil to the R2A solution. With the R2A- bacteria
broth from the second foil, we made serial dilutions and spread plates. We planned to
create serial dilutions for up to 1x10-8. Each serial dilution tube contained 0.9 of R2A
and 0.1 of the bacteria-R2A broth would later be added in for dilution. We transferred

56 | P a g e

0.1mL of the original R2A- bacteria broth into a tube contain 0.9mL of R2A. This tube was
labeled 1x10-1. After using the vortex to mix thoroughly, we transferred 0.1 from the
tube labeled 1x10-1 to a tube labeled 1x10-2. We continued this all the way until we reached
1x10-8. After creating all serial dilutions, we used a cell spreader and Bunsen burner to
create spread plates starting from 1x10-3 until 1x10-8. Finally we Para filmed all plates and
allowed multiple days for bacterial growth at room temperature.

Although looking under a microscope at a hemocytometer is not the best method for

quantification, we did for our project. This served as another method, along with the serial
dilutions and the press plate, to have data. We performed four trials total. Trial 1 was done
here at E.B. Doran for 2.5 . The incubation time was for about 50 . Trials 2-4
were done in Maringouin, LA which is about 20 west of Baton Rouge. For these three
trials, the temperature and humidity sensors were active during the flights. Trial 2 had a
!"##$

cell concentration count of 5.4X 108 !" , a collection time of 2.5 , an incubation
time of 26.5 , a temperature reading of 30.4C 1.6, and a relative humidity of 63.3%
!"##$

3.8. Trial 3 had a cell concentration of 4.7x108 !" , a collection time of 2.5 , an
incubation time of 24 , a temperature reading of 27.3C 0.5, and a relative humidity
of 77.7% 1.6. Last, trial 4 had a cell concentration of 9.9x108 /, a collection time of
5 , an incubation time of 24 , a temperature of 25.2C 0.2, and a relative
humidity of 84.9% 2.6. We do know that in order to have statistically correct bacterial
analysis we needed to have collected three times within the same conditions. Although we
did not do this, when comparing trials 3 and 4, there is about two times the cell
concentration on trial 4 than trial 3 with trail 4 having twice the flight time. This data
shows that our bacterial collection was precise.

57 | P a g e

Figure 23: Hemocytometer view

of Trial 1.

Figure 24: Hemocytometer view

of Trial 2.

Figure 25: Hemocytometer view

of Trial 3.

Figure 26: Hemocytometer view

of Trial 4.

Redesign:

Since the group is in the process of patenting the ACROBE, we have decided not to

give specifics to the design. However, we do know that we need to have a more efficient
hexacopter that can have better batteries to have a longer flight time and carry a heavier
load.

58 | P a g e

Schedule:

Budget:
Vendor
Amazon











Hobby Towne



Lowes

Figure 27: This Gantt Chart contains deadlines for Spring 2015
semester.

Items
Polycarbonate tubing 1/4 ID
Polycarbonate tubing 3/4 ID
Polycarbonate tubing 7/8 ID
Polycarbonate tubing 1" ID
Polycarbonate tubing 1/2" ID
Polycarbonate tubing
actron vacuum and fuel pressure kit
polycarbonate tubing 3/4 ID
Polycarbonate tubing
Temperature and Humidty Sensor
RC Lipo Battery
Voltage Checker
Epoxy
nuts, bolts, balsa woodetc
Wood
monocoat
1-inx 5ft Sch-40 PI
3/4 in x 5 ft Sch40 PVC

Price
$20.25
$33.65
$16.11
$13.67
$25.09
$19.71
$17.84
$29.14
$29.74
$29.95
$22.99
$5.60
$15.99
$22.30
$39.51
$28.27
$3.09
$1.79
59 | P a g e













Digi-Key

Pololu
The Felt Store
The Composite Store




Hobby King

Dollar Tree



Hobby Lobby
Online Metals
Adafruit
Mouser
Spark Fun

Radio Shack

Total with Shipping

2 in x 2ft PVC SCh 40 SOL

2 in sch 40 cap socket
1/2 in x 2 ft PVC Sch 40
1/2 in Sch40 cap
1 in sch40 elbow (3)
1 in sch40 coupling(2)
3/4 in sch40 elbow (3)
3/4 in sch40 adapter
3/4 in sch40 adapter
1 in sch40 adapter
1 in x 3/4 in adapter
8x10 clear lexan (3)
ABS Pressure Sensor (x3)
IC encoder ABS magnetic
Continuous Rotation Servo
Neoprene
Carbon Fiber Tube (x2) 4 ft long
Resin
Hardener
Carbon Fiber Tube 2 ft long (x2)
Carbon Fiber Fabric (x2)
Digital Air Speed Sensor & Pitot Tube
Set
Woven Carbon Fiber Sheet (x3)
Mxing Bowl
Foos\d Storage Set
Cars Tumbler
Snack Cont
Mold Kit
Aluminum Bars
Slip Ring
Arduino Mega Proto PCB (x2)
Sparkfun Open Log
SD Card
Pins

$4.14
$2.08
$3.03
$0.97
$1.98
$0.92
$1.41
$1.86
$0.47
$0.77
$1.56
$12.72
$48.27
$39.86
$17.90
$29.94
$8.70
$43.35
$21.32
$5.00
$57.17
$49.44
$68.10
$1.00
$1.00
$1.00
$1.00
$6.99
$67.63
$29.13
$16.89
$28.96
$17.96
$5.43

$1,054.70

Table 9: Final budget at the conclusion of the Spring 2015 semester.

60 | P a g e


Bill of Materials:

Item
Temperature and Humidity
Sensor
Pressure Sensor
Continuous Rotation Servo
Carbon Fiber Tubing (4 ft)
Carbon Fiber Tubing (2 ft)
Resin
Hardener
Epoxy
Rotary Encoder
Woven Carbon Fiber Sheet
Slip Ring
Arduino Mega Proto PCB
Sparkfun Logger
SD Card
Carbon Fiber Fabric
RC Lipo Battery
Arduino Mega
22 ga. Insulated Wire (10 ft)
Neodymium Magnets
Futaba Receiver
Electronic Switch
2.5 mm nuts/bolts
Resistors and Capacitors

Quantity

Total
1
$29.95

2
1
2
2
1
1
1
1
5
1
1
1
1
2
1
1
1
8
1
1
20

$32.18
$17.90
$8.70
$5.00
$43.35
$21.32
$15.99
$39.86
$68.10
$29.13
$8.45
$28.96
$17.96
$57.17
$22.99
$37.03
$6.99
$5.00
$69.98
$7.39
$10.00
$10.00

2 each

Table 10: Complete Bill of Materials.

61 | P a g e

Conclusion:
In conclusion, the scope of this senior design project was able to be completed
underneath the umbrella of Biological Engineering. The engineering principles learned
throughout the course of our undergraduate years here at LSU helped us achieve our goals.
We were able to accomplish our goals/objectives of making sure the ACROBE is modular,
easy to use, capable of multiple runs in a day, and be autonomous.

62 | P a g e

References:
[1] Christner, Brent C. "Cloudy with a Chance of Microbes." News Magazine of the American
Society for Microbiology Feb. 2012: 70-75. Web. 20 Nov. 2014.

[2] Smith, A., and R. Katz, 2013: U.S. Billion-dollar Weather and Climate Disasters: Data

Sources, Trends, Accuracy and Biases. Natural Hazards.

[3] Granger, Doug. "High Altitude Student Payload (HASP)." LSU Space Sciences Group.

Louisiana State University, n.d. Web.

[4] Journal of Microbiological Methods, Volume 107, December 2014, Pages 161-168 N.C.

[5] Whyte, W., G. Green, and A. Albisu. "Collection Efficiency and Design of Microbial Air
Samplers." Journal of Aerosol Science 38.1 (2007): 97-110.ScienceDirect. Web. 15
Sept. 2014

[6] Carbon Fiber Fabric." 3K Plain Weave Carbon Fiber. The Composite Store (CST), 2014.
Web. 02 Dec. 2014.

[7] "West Systems Laminating Epoxy Resin." West System Epoxy. The Composite Store
(CTS), 2014. Web. 02 Dec. 2014.

[8] "6061 Aluminum." 6061 Aluminum, Aluminium, Alumina Bobco Metals LLC. Bob Co.
Metals, 2014. Web. 02 Dec. 2014.

[9] "Nylon Nuts, Bolts & Washers." IndustrySearch. Industry Search, 2014. Web. 02 Dec.
2014.
[10] Dulbecco, R. et al. (1954): Plaque formation and isolation of pure lines with poliomyelitis
viruses. In: J. Exp. Med. vol. 99 (2), pp. 167-182.
[11] Reasoner, D. J., and E. E. Geldreich. 1979. A new medium for the enumeration and

subculture of bacteria from potable water.
[12] Boundless. Viable Cell Counting. Boundless Microbiology. Boundless, 14 Nov. 2014.
[13] Wolochow, H. The Membrane Filter Technique for Estimating Numbers of Viable
Bacteria: Some Observed Limitations with Certain Species. Applied Microbiology 6.3
(1958): 201206. Print.
[14] Sauer S., Kliem M. 2010. Mass spectrometry tools for the classification and
identification of bacteria. Nat. Rev. Microbiol. 8:7482.

63 | P a g e