vii
TABLE OF CONTENTS
CHAPTER
1
2
TITLE
PAGE
DECLARATION
ii
DEDICATION
iii
ACKNOWLEDGMENT
iv
ABSTRACT
v
ABSTRAK
vi
TABLE OF CONTENTS
vii
LIST OF TABLES
x
LIST OF FIGURES
xi
LIST OF ABBREVIATIONS
xiv
LIST OF SYMBOLS
xv
INTRODUCTION
1.1
Introduction
1
1.2
Problem Statement
3
1.3
Project Objectives
5
1.4
Scope of the Project
5
1.5
Summary of the Chapter
6
LITERATURE REVIEW
2.1
Devices used in microbial cultivation
7
2.1.1 Bench scale Bioreactor
7
2.1.2
Shake flask culture
9
2.1.3
Microbioreactor
12
2.1.4
Comparison
14
viii
2.2
2.3
Considerations in microbioreactor design
17
2.2.1
Fabrication materials
17
2.2.2
Mixing
18
2.2.3
Temperature
19
2.2.4
pH
21
2.2.5
Evaporation
22
Previous works on microbioreactor
2.3.1
Development of a single-use microbioreactor by
D. Schäpper el. al (2010)
2.3.2
3
23
Development of a multiplexed microbioreactor
system by Szita N el. al (2005)
2.4
23
Summary of the Chapter
28
33
DESIGN AND DEVELOPMENT
3.1
Introduction
34
3.2
Hardware development
35
3.2.1
Temperature Measurement
35
3.2.1.1 The LM35
35
3.2.1.2 The PT100
37
3.2.2
Speed Measurement
46
3.2.3
Actuators
52
3.2.3.1 Motor
52
3.2.3.2 Heater
53
DAQ
55
3.2.4
3.3
3.4
Software Development
56
3.3.1
Arduino Software
56
3.3.2
GUI
57
3.3.2.1 Setting up the interface
58
3.3.2.2 Extra Considerations
60
3.3.2.3 Simple Example
64
3.3.3
PI Controller
65
3.3.4
PCB design
66
Summary of the Chapter
68
ix
4
RESULTS AND DISCUSSION
4.1
Introduction
69
4.2
First phase
70
4.2.1
Hardware
70
4.2.2
Temperature control
71
4.2.3
Speed control
73
4.3
5
Second phase
75
4.3.1
Hardware
75
4.3.2
Temperature control
77
4.3.3
Speed control
78
4.4
LabView GUI
82
4.5
Summary of the Chapter
87
CONCLUSION AND FUTURE PLAN
5.1
Conclusion
88
5.2
Future plan
89
REFERENCES
90
Appendix A
93
x
LIST OF TABLES
TABLE NO.
TITLE
PAGE
2.1
Comparison
15
3.1
PT100 Chart of values
38
3.2
PI parameters
65
4.1
Testing PI parameters
79
xi
LIST OF FIGURES
FIGURE NO
TITLE
PAGE
1.1
New Brunswick bioreactor
3
1.2
Prototype microbioreactor with 100 μl working volume
3
2.1
Bench scale bioreactor
9
2.2
Shake flask
10
2.3
Shake flask shaker
10
2.4
Shake flask culture
11
2.5
Controlled environment chamber
12
2.6
Microbioreactor platform at UTM
13
2.7
The microbioreactor mechanical design
13
2.8
Mixing scheme through the mounting of stirrer bar on a revolting
central shaft
19
2.9
The PT100 Temperature sensor
20
2.10
Photo of the complete microbioreactor seen from the bottom
24
2.11
Sketch of both main reactor layers as seen from below
24
2.12
Mixing in the microbioreactor at 500rpm
26
2.13
Temperature response at different set points
27
2.14
Displaying set points and the measured values of pH
28
2.15
The reactor design
29
2.16
Photograph of the multiplexed microbioreactor system
29
2.17a
Parallel fermentations of E. coli
31
2.17b
Parallel fermentations of E. coli
31
2.18
Comparison with bench scale bioreactor
32
3.1
LM35 terminal connections
35
3.2
Circuit schematic of the LM35 followed by the amplifier
36
xii
3.3
PT100
37
3.4
Transmitter
39
3.5
Wheatstone bridge
39
3.6
Difference amplifier
41
3.7
Non-inverting operational amplifier
42
3.8
Low-Pass filter
43
3.9
Conditioning circuit for the PT00
44
3.10
Simulation of PT100 circuit in Protus
45
3.11
Hall-effect sensor
46
3.12
Terminal connections
46
3.13
ATMEGA328P Pins mapping
47
3.14
Complete Circuitry for speed measurement
48
3.15
Flow chart for speed measurement
50
3.16
Stages of development
53
3.17
Control circuit
54
3.18
Arduino mega 2560
55
3.19
Arduino IDE
56
3.20
Firmware for Arduino
59
3.21
Arduino Functions in LabView
59
3.22
Init VI
61
3.23
Init Front Panel
61
3.24
Init Block Diagram
62
3.25
Example
64
3.26
First Design
66
3.27a
Second design main board
67
3.27b
Expansion to the second design
68
4.1
Hardware of the first stage
70
4.2
Temperature control response
71
4.3
Testing condition where the heater placed underneath
71
4.4
Temperature control response
72
4.5
Testing condition where the heaters placed on the side
73
4.6
Speed control response
74
4.7
Main board
75
4.8
Optional expansion
76
xiii
4.9
Temperature control response
77
4.10
Testing condition where the heater placed underneath
77
4.11
Speed control response
78
4.12
Testing condition
78
4.13
Speed control response for set one
79
4.14
Speed control response for set two
80
4.15
Mixing Quality test at 300rpm
81
4.16
Block diagram for temperature control
82
4.17
Front Panel for temperature control
83
4.18
Block diagram for speed control
84
4.19
Front Panel for speed control
85
4.20
Block diagram of the combined GUI
86
4.21
Front panel of the combined GUI
86
4.22
Testing conditions
87
xiv
LIST OF ABBREVIATIONS
DAQ
-
Data Acquisition Card
DO
-
Dissolved Oxygen
OD
-
Optical Density
NI
-
National Instruments
PID
-
proportional-integral-derivative
GUI
-
Graphical User Interface
rpm
-
revolution per minute
PMMA
-
polymethylmethacrylate
PDMS
-
polydimethylsiloxane
RTD
-
resistance temperature detectors
S/V
-
surface area to working volume ratio
ISFET
-
ion-sensitive and field-effect transistor
PWM
-
Pulse-width modulation
ADC
-
Analog Digital converter
PCB
-
printed circuit board
IC
-
integrated circuit
xv
LIST OF SYMBOLS
mL
-
millilitre
µL
-
microlitre
°C
-
celsius
V
-
voltage/volts
mV
-
millivolt
µV
-
microvolt
A
-
ampere
mA
-
milliampere
Ω
-
ohms
µF
-
microfarad
Hz
-
hertz