Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
xi
1 Culturing Mycobacteria. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
Elizabeth Wallace, Debra Hendrickson, Nicholas Tolli, Carolina Mehaffy,
Marı ´a Pen ˜a, Jerry A. Nick, Phillip Knabenbaur, Jackson Watkins,
Anne Simpson, Anita G. Amin, Delphi Chatterjee, Karen M. Dobos,
Ramanuj Lahiri, Linda Adams, Michael Strong, Max Salfinger,
Rebecca Bradford, Timothy T. Stedman, Marco A. Riojas,
and Manzour Hernando Hazb on
2 DNA Isolation from Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Heena Jagatia and Daire Cantillon
3 Extraction and Separation of Mycobacterial Proteins . . . . . . . . . . . . . . . . . . . . . . . . 77
Megan Lucas, Joan M. Ryan, Jackson Watkins, Kala Early,
Nicole A. Kruh-Garcia, Carolina Mehaffy, and Karen M. Dobos
4 Lipid and Lipoarabinomannan Isolation and Characterization . . . . . . . . . . . . . . . . 109
Marie-Antoinette Lane ´elle, Lucie Spina, Je ´ro ˆme Nigou,
Anne Lemassu, and Mamadou Daffe ´
5 Purification of Hibernating and Active CÀ Ribosomes
from Zinc-Starved Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
Yunlong Li, Pooja Keshavan, Jamie H. Corro, Ravi K. Koripella,
Rajendra K. Agrawal, and Anil K. Ojha
6 Macrophage Infection Models for Mycobacterium tuberculosis . . . . . . . . . . . . . . . . 167
Benjamin K. Johnson, Sean M. Thomas, Andrew J. Olive,
and Robert B. Abramovitch
7 Novel Single-Cell and High-Throughput Microscopy Techniques
to Monitor Dictyostelium discoideum–Mycobacterium marinum
Infection Dynamics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Manon Mottet, Cristina Bosmani, Nabil Hanna,
Jahn Nitschke, Louise H. Lefranc ¸ois, and Thierry Soldati
8 Single-Cell Analysis of Mycobacteria Using Microfluidics
and Time-Lapse Microscopy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
Giulia Manina and Neeraj Dhar
9 Measuring Efflux and Permeability in Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . . . 231
Liliana Rodrigues, Jose ´ A. Aı ´nsa, and Miguel Viveiros
10 Drug Susceptibility Screening Using In Vitro Models
of Hypoxic Non-Replicating Persistent Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . 247
Savannah E. R. Gibson, James Harrison, and Jonathan A. G. Cox
11 Flow Cytometry Analysis of Mycobacteria and Mycobacteria-Infected
Immune Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261
Sydney L. Solomon and Bryan D. Bryson
vii
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
xi
1 Culturing Mycobacteria. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
Elizabeth Wallace, Debra Hendrickson, Nicholas Tolli, Carolina Mehaffy,
Marı ´a Pen ˜a, Jerry A. Nick, Phillip Knabenbaur, Jackson Watkins,
Anne Simpson, Anita G. Amin, Delphi Chatterjee, Karen M. Dobos,
Ramanuj Lahiri, Linda Adams, Michael Strong, Max Salfinger,
Rebecca Bradford, Timothy T. Stedman, Marco A. Riojas,
and Manzour Hernando Hazb on
2 DNA Isolation from Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Heena Jagatia and Daire Cantillon
3 Extraction and Separation of Mycobacterial Proteins . . . . . . . . . . . . . . . . . . . . . . . . 77
Megan Lucas, Joan M. Ryan, Jackson Watkins, Kala Early,
Nicole A. Kruh-Garcia, Carolina Mehaffy, and Karen M. Dobos
4 Lipid and Lipoarabinomannan Isolation and Characterization . . . . . . . . . . . . . . . . 109
Marie-Antoinette Lane ´elle, Lucie Spina, Je ´ro ˆme Nigou,
Anne Lemassu, and Mamadou Daffe ´
5 Purification of Hibernating and Active CÀ Ribosomes
from Zinc-Starved Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
Yunlong Li, Pooja Keshavan, Jamie H. Corro, Ravi K. Koripella,
Rajendra K. Agrawal, and Anil K. Ojha
6 Macrophage Infection Models for Mycobacterium tuberculosis . . . . . . . . . . . . . . . . 167
Benjamin K. Johnson, Sean M. Thomas, Andrew J. Olive,
and Robert B. Abramovitch
7 Novel Single-Cell and High-Throughput Microscopy Techniques
to Monitor Dictyostelium discoideum–Mycobacterium marinum
Infection Dynamics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Manon Mottet, Cristina Bosmani, Nabil Hanna,
Jahn Nitschke, Louise H. Lefranc ¸ois, and Thierry Soldati
8 Single-Cell Analysis of Mycobacteria Using Microfluidics
and Time-Lapse Microscopy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
Giulia Manina and Neeraj Dhar
9 Measuring Efflux and Permeability in Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . . . 231
Liliana Rodrigues, Jose ´ A. Aı ´nsa, and Miguel Viveiros
10 Drug Susceptibility Screening Using In Vitro Models
of Hypoxic Non-Replicating Persistent Mycobacteria . . . . . . . . . . . . . . . . . . . . . . . 247
Savannah E. R. Gibson, James Harrison, and Jonathan A. G. Cox
11 Flow Cytometry Analysis of Mycobacteria and Mycobacteria-Infected
Immune Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261
Sydney L. Solomon and Bryan D. Bryson
vii