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 Handbook of
Full-Field Optical Coherence Microscopy
 Handbook of
Full-Field Optical Coherence Microscopy
Technology and Applications

edited by
Arnaud Dubois
May 13, 2016 10:35 PSP Book - 9in x 6in 00-Arnaud-Dubois-Prelims

Published by
Pan Stanford Publishing Pte. Ltd.
Penthouse Level, Suntec Tower 3
8 Temasek Boulevard
Singapore 038988

Email: editorial@panstanford.com
Web: www.panstanford.com

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library.

Handbook of Full-Field Optical Coherence Microscopy: Technology

and Applications
Copyright 
c 2016 Pan Stanford Publishing Pte. Ltd.

All rights reserved. This book, or parts thereof, may not be reproduced in any
form or by any means, electronic or mechanical, including photocopying,
recording or any information storage and retrieval system now known or to
be invented, without written permission from the publisher.

For photocopying of material in this volume, please pay a copying

fee through the Copyright Clearance Center, Inc., 222 Rosewood Drive,
Danvers, MA 01923, USA. In this case permission to photocopy is not
required from the publisher.

ISBN 978-981-4669-16-0 (Hardcover)

ISBN 978-981-4669-17-7 (eBook)

Printed in the USA

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Contents

Preface xix

1 Introduction to Full-Field Optical Coherence Microscopy 1

Arnaud Dubois
1.1 Introduction 1
1.2 Experimental Setup 4
1.2.1 General Principle 4
1.2.2 Interference Microscope Configuration 5
1.2.3 Light Source 6
1.2.4 Image Sensor 11
1.3 Image Acquisition 13
1.3.1 Axial Scan 13
1.3.2 Phase-Shifting Interferometry 15
1.3.3 Sinusoidal Phase Shifting and Four Integrating
Buckets 18
1.4 Performance 22
1.4.1 Detection Sensitivity 22
1.4.2 Axial Resolution 25
1.4.3 Lateral Resolution 29
1.4.4 Sample Motion Artifacts 30
1.5 Clinical Applications 31
1.5.1 Ophthalmic Tissue Imaging 32
1.5.2 Skin Tissue Imaging 33
1.5.3 Breast Tissue Imaging 34
1.5.4 Other Tissue Imaging 36
1.6 Applications in Biology 37
1.6.1 Developmental Biology 37
1.6.2 Cell Biology 38
1.6.3 Plant Studies 39
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vi Contents

1.7 Material Characterization 39

1.8 Conclusion 40

PART I THEORETICAL ASPECTS

2 Theory of Imaging and Coherence Effects in Full-Field Optical

Coherence Microscopy 53
Anton A. Grebenyuk and Vladimir P. Ryabukho
2.1 Introduction 54
2.2 Signal Formation in FF-OCM 55
2.2.1 Principle of the Analysis 56
2.2.2 Optical Transmission Functions of the Sample
and Reference Arms 61
2.2.3 Resultant Signal of FF-OCM 66
2.3 Coherence Effects in FF-OCM 69
2.3.1 Principle of Coherence Gating 70
2.3.1.1 Sample with uniform interfaces 70
2.3.1.2 Sample with transversal structure 73
2.3.2 Impulse Response and Illumination Aperture
Function 76
2.3.3 Influence of the Sample Refractive Index 80
2.4 Conclusions 83

3 Spatiotemporal Coherence Effects in Full-Field Optical

Coherence Tomography 91
Ibrahim Abdulhalim
3.1 Introduction 91
3.2 Basic Coherence Concepts 94
3.3 Finite Temporal and Infinite Longitudinal Spatial
Coherence Lengths 97
3.4 Finite Longitudinal Spatial and Infinite Temporal
Coherence Lengths 101
3.5 Lateral Mismatch as a Source of Lateral Decoherence 105
3.6 Finite Longitudinal and Finite Temporal Coherence
Lengths 107
3.7 Effects on Multilayer Sample Measurement 111
3.7.1 LSC Effect on the Fringe Size 111
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Contents vii

3.7.2 LSC Effect on the Thickness Determination

When Imaging a Multilayered Sample 112
3.7.3 LSC Effect on the Imaging Depth 115
3.8 Conclusions and Future Trends 119

4 Cross Talk in Full-Field Optical Coherence Tomography 131

Boris Karamata, Marcel Leutenegger, and Theo Lasser
4.1 Optical Cross Talk in FF-OCT 135
4.1.1 Origin and Definition of Cross-Talk Noise 136
4.1.2 Qualitative Analysis of the Cross-Talk
Contribution 137
4.1.2.1 Spatial coherence of cross talk light 138
4.1.2.2 Interference with multiply scattered
light 139
4.1.2.3 Questions raised by our analyses 140
4.1.3 Cross-Talk Noise Suppression with Spatial
Coherence Gating 141
4.2 Theory and Model 143
4.2.1 Mathematical Description 144
4.2.2 Calculation of the Mean Signal 148
4.2.3 Integration of Monte Carlo Results 151
4.3 Method for Cross-Talk Investigation 151
4.3.1 Setup 152
4.3.2 Sample 154
4.3.3 Presentation of Experimental Results 155
4.3.4 Monte Carlo Simulation Details 157
4.4 Experimental and Theoretical Results for Cross-Talk
Noise 158
4.4.1 Dependence on Sample Properties 158
4.4.2 Cross-Talk Contribution Relative to the Useful
Signal 161
4.4.3 Additional Results and Conclusions 164
4.5 Cross-Talk Suppression 164
4.5.1 Setup and Sample 165
4.5.2 Results 168
4.6 Conclusions and Discussions 170
4.6.1 Full-Field OCT 170
4.6.2 Modeling Multiple Scattering in OCT 173
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viii Contents

5 Signal Processing Methods in Full-Field Optical Coherence

Microscopy 183
Igor Gurov
5.1 Introduction 183
5.2 Basics of Fringe Signal Registration 186
5.3 TD-OCT and FD-OCT Information Capacity 189
5.3.1 Information Channel and Capacity 190
5.3.2 2D and 3D OCT Imaging 193
5.4 Signal Downsampling in Full-Field OCT 194
5.4.1 Spectral Characteristics of Downsampled
Signals 197
5.5 Dynamic Fringe Signal Evaluation in State Space 203
5.5.1 Recurrence Computational Algorithms Applied
to Fringe Processing 206
5.5.2 Application of Recurrence Processing
Algorithm in State Space to Low-Coherence
Profilometry 207
5.5.3 Application of Recurrence Processing
Algorithm in State Space When Evaluating
Random Tissues 207
5.5.4 Combined Use of Signal Downsampling and
Kalman Filtering 210
5.5.5 Recurrence Filtering Modifications and
Processing Speed 211
5.6 Recurrence Processing of Swept-Source Spectral
Interference Fringes 212
5.7 Discussion and Conclusions 217

PART II TECHNOLOGICAL DEVELOPMENTS

6 High-Speed Image Acquisition Techniques of Full-Field

Optical Coherence Tomography 223
Byeong Ha Lee, Woo June Choi, Kwan Seob Park,
and Gihyeon Min
6.1 Introduction 223
6.2 High-Speed 2D Imaging of Full-Field Optical
Coherence Tomography 224
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Contents ix

6.2.1 Phase-Stepping Method Based on Hilbert

Transformation 224
6.2.2 Integrating Bucket Method Based on Hilbert
Transformation 228
6.2.3 Quadrature Fringes Wide-Field Optical
Coherence Tomography 230
6.2.4 Single-Shot Phase-Stepping Methods 234
6.3 High-Speed 3D Imaging of Full-Field Optical
Tomography 242
6.3.1 2D Heterodyne Detection with a Pair of CCD
Cameras 242
6.3.2 2D Heterodyne Detection with a Single CCD
Camera 248
6.3.3 2D Heterodyne Detection with a Single CCD
Camera and an Active Feedback Loop 255
6.4 Summary 261

7 Toward Single-Shot Imaging in Full-Field Optical Coherence

Tomography 267
Bettina Heise
7.1 Introduction 267
7.2 From Phase Shifting toward Single-Shot Techniques 269
7.2.1 Temporal Phase-Shifting Techniques 270
7.2.2 Simultaneous Phase-Shifting Techniques 273
7.2.2.1 Dual-shot image acquisition 273
7.2.2.2 Single-shot image acquisition 274
7.2.2.3 Polarization-based schemes 275
7.2.2.4 Unpolarized instantaneous
phase-shifting schemes 276
7.2.2.5 Heterodyne detection schemes 277
7.2.2.6 SLM-based schemes 278
7.2.3 Hyperspectral Single-Shot Techniques 279
7.3 Signal Processing Methods 281
7.3.1 Phase-Stepping Approaches 281
7.3.2 The Subtraction Method 282
7.3.3 Analytic Signal–Based Reconstruction 283
7.4 Applications 289
7.5 Summary and Outlook 295
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x Contents

8 Frequency Domain Full-Field Optical Coherence Tomography 303

Rainer A. Leitgeb, Abhishek Kumar, and Wolfgang Drexler
8.1 Introduction 303
8.2 General Concept 305
8.2.1 Setup and System Components 305
8.2.2 Standard SS-OCT Signal Processing 308
8.2.3 Sensitivity 311
8.3 Digital Refocusing 313
8.4 Outlook 317

9 Full-Field OCM for Endoscopy 323

Anne Latrive and Claude Boccara
9.1 Introduction 324
9.1.1 Overview of Endoscopy 324
9.1.2 Endoscopic OCT 325
9.1.2.1 OCT for flexible endoscopy 325
9.1.2.2 OCT for rigid endoscopy 326
9.1.3 Problematic of Endoscopic Full-Field OCM 326
9.2 Theory of Full-Field OCM with Two Coupled
Interferometers 327
9.2.1 Dual-Interferometer OCT Signal 327
9.2.2 Performances 330
9.2.2.1 Sensitivity 330
9.2.2.2 Transversal resolution 331
9.2.2.3 Axial resolution 332
9.2.3 Conclusion 333
9.3 Problematics of Endoscopic Imaging with a Flexible
Fiber Bundle 334
9.3.1 Pixelation Effect Removal 334
9.3.1.1 Fourier domain filters 334
9.3.1.2 Deconvolution 335
9.3.1.3 Interpolation between the cores 336
9.3.2 Cross-Talk Interferences 337
9.3.2.1 Core-to-core coupling 337
9.3.2.2 Mode-to-mode coupling 340
9.3.3 Conclusion 341
9.4 Full-Field OCM with a Flexible Endoscope 341
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Contents xi

9.4.1 FF-OCM System with Simple Imaging

Interferometer and SLD Source 342
9.4.2 FF-OCM System with Complex Imaging
Interferometer and Xenon Arc Source 343
9.4.3 FF-OCM System with Simple Imaging
Interferometer and Xenon Arc Source 345
9.4.4 Discussion 346
9.5 Full-Field OCM with a Rigid Endoscope 346
9.5.1 Choice of the FF-OCM Setup 347
9.5.2 FF-OCM Setup with a Rigid Probe 347
9.5.2.1 Setup and performances 347
9.5.2.2 Imaging results 348
9.5.3 Discussion 349
9.6 Conclusion 351

10 Full-Field Optical Coherence Tomography and Microscopy

Using Spatially Incoherent Monochromatic Light 357
Dalip Singh Mehta, Vishal Srivastava, Sreyankar Nandy,
Azeem Ahmad, and Vishesh Dubey
10.1 Introduction 357
10.2 Principle of Low Coherence Interferometry Based
on Temporal Coherence of the Light Source 360
10.2.1 The Wiener–Khintchine Theorem of
Temporal Coherence 362
10.2.2 Resolution of OCT Systems Based on
Temporally Low Coherent Light 365
10.3 Principle of Spatial Coherence Gated Tomography
with a Monochromatic Light Source 368
10.3.1 Transverse (Lateral) Spatial Coherence of
Light Vibrations 370
10.3.2 Longitudinal Spatial Coherence of Light
Vibrations 373
10.4 Principle of Spatial Coherence Gated Microscopy
with Monochromatic Light 376
10.5 Experimental Details for Spatial Coherence Gated
Tomography with Monochromatic Light 377
10.5.1 Determination of Longitudinal Spatial
Coherence Length 379
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xii Contents

10.5.2 Application of Longitudinal Spatial

Coherence in Profilometry 381
10.5.3 High-Resolution Longitudinal Spatial
Coherence Gated Tomography 382
10.6 Coherence Holography 385
10.7 Conclusion 386

11 Real-Time and High-Quality Online 4D FF-OCT Using

Continuous Fringe Scanning with a High-Speed Camera and
FPGA Image Processing 393
P. C. Montgomery, F. Anstotz, D. Montaner, and F. Salzenstein
11.1 Introduction 394
11.2 Theory 398
11.2.1 Interference Microscopy 398
11.2.2 Modeling of the Optical Probe in
Interference Microscopy 400
11.3 Envelope Detection Algorithms 402
11.3.1 Peak Fringe Detection Algorithm 403
11.3.2 The Fringe Modulation Algorithm 404
11.4 Experimental 407
11.5 System Performance 412
11.6 Applications 415
11.6.1 Laterally Moving Microfluxgate Surface
Measured with the PFSM Algorithm 415
11.6.2 Laterally Moving Microfluxgate Surface
Measured with the FSA Algorithm 417
11.6.3 Laterally Moving GaN Surface Measured
with the FSA Algorithm 417
11.6.4 Drying Drop of Liquid Correction Whitener
Measured with the FSA Algorithm 419
11.7 Future Developments and Potential Applications in
FF-OCT 420
11.8 Conclusions 423

12 Digital Interference Holography for Tomographic Imaging 429

Lingfeng Yu, Mariana C. Potcoava, and Myung K. Kim
12.1 Introduction 429
12.2 Principle of Digital Interference Holography 432
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Contents xiii

12.2.1 Basic Description of DIH 432

12.2.2 Phase Correction in DIH 433
12.2.3 Spectral Shaping in DIH 434
12.3 Techniques of DIH 437
12.3.1 Detailed Description of DIH Experiments 437
12.3.2 DIH by the Angular Spectrum Algorithm 439
12.3.3 Variable Tomographic Scanning 442
12.3.4 DIH Based on Spectral Interferometry 445
12.4 Applications of DIH 450
12.4.1 Animal Tissue 450
12.4.2 Human Retina 451
12.4.3 DIH for Biometric Application 454
12.4.4 Submicron Tomography of Cells with DIH 457
12.5 Discussions 457

PART III ADDITIONAL IMAGING MODALITIES

13 Technological Extensions of Full-Field Optical Coherence

Microscopy for Multicontrast Imaging 467
Arnaud Dubois
13.1 Introduction 467
13.2 Polarization-Sensitive FF-OCM 469
13.2.1 Experimental Setup and Principle 470
13.2.2 Image Calculation 472
13.2.3 Validation 476
13.3 Spectroscopic FF-OCM 479
13.3.1 Experimental Setup 479
13.3.2 Intensity-Based Tomographic Imaging 480
13.3.3 Spectroscopic Measurements 481
13.3.4 Interpretation of Spectroscopic
Measurements 482
13.3.5 Validation of the Spectroscopic
Measurements 484
13.3.6 Demonstration of Imaging Contrast
Enhancement 486
13.3.7 Spectroscopic Polarization-Sensitive
FF-OCM 487
13.4 Multispectral FF-OCM 490
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xiv Contents

13.4.1 Dual-Band FF-OCM 491

13.4.1.1 Method 491
13.4.1.2 Simultaneous dual-band imaging 494
13.4.2 Three-Band FF-OCM 498
13.4.2.1 Materials and methods 498
13.4.2.2 System characteristics and image
results 501
13.5 Combination of FF-OCM with Fluorescence
Microscopy 505
13.5.1 Materials and Methods 506
13.5.2 System Performance and Image Results 508
13.6 Conclusion 510

14 Spectroscopic Full-Field Optical Coherence Tomography 519

Julien Moreau
14.1 FF-OCT Principle 520
14.2 Spectroscopic FF-OCT 524
14.3 Limitations of Spectroscopic FF-OCT 530

15 Multiwavelength Full-Field Optical Coherence Tomography 533

Mariana C. Potcoava, Nilanthi Warnasooriya, Lingfeng Yu,
and Myung K. Kim
15.1 Introduction 533
15.2 Basic Single-Wavelength, Low-Coherence
Interferography 536
15.2.1 Digital Focusing in Low-Coherence
Interferometry 538
15.3 Multiwavelength, Optical-Phase-Unwrapping,
Low-Coherence Interferography 544
15.3.1 Two-Wavelength Optical Phase
Unwrapping 547
15.3.2 Three-Wavelength Optical Phase
Unwrapping 548
15.3.3 Application for Two-Wavelength Optical
Phase Unwrapping 549
15.3.4 Application for Three-Wavelength Optical
Phase Unwrapping 551
15.4 Full-Color FF-OCT 553
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Contents xv

15.4.1 Results of Full-Color FF-OCT 556

15.5 Discussion 559

16 Dual-Modality Full-Field Optical Coherence and

Fluorescence Sectioning Microscopy: Toward All Optical
Digital Pathology on Freshly Excised Tissue 565
Fabrice Harms
16.1 Introduction: Clinical Context 565
16.2 Optical Coherence Tomography in Pathology: Early
Multimodal Approaches and Limitations 568
16.2.1 Applying OCT to Pathology Assessment of
Biological Tissue 568
16.2.2 High-Resolution OCT for Pathology:
Full-Field Optical Coherence Tomography 569
16.2.3 Multimodal OCT Approaches 573
16.3 Multimodal Full-Field Optical Coherence
Tomography for Pathology Applications 574
16.3.1 Translating FF-OCT to Pathology Diagnosis 574
16.3.2 Combined FF-OCT and Fluorescence
Sectioning Microscopy 576
16.3.2.1 Setup 576
16.3.2.2 Full-field optical coherence
tomography: resolution 578
16.3.2.3 Structured illumination
fluorescence microscopy 579
16.3.3 Multimodal FF-OCT/Fluorescence Images
of Healthy and Cancerous Tissue Samples 582
16.3.3.1 Sample selection and preparation 582
16.3.3.2 Imaging protocol 583
16.3.3.3 Results 584
16.4 Discussion and Conclusion 589

PART IV APPLICATIONS

17 Full-Field Optical Coherence Tomography for Rapid

Histological Evaluation of ex vivo Tissues 595
Manu Jain and Sushmita Mukherjee
17.1 Introduction 595
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xvi Contents

17.2 Current Practice for Intraoperative Diagnosis and

Its Limitations 596
17.3 Basic Principles and Instrumentation of FF-OCT 597
17.4 FF-OCT-Generated Histology Atlas of Rat Organs 598
17.4.1 Skin 599
17.4.2 Stomach 600
17.4.3 Liver 600
17.4.4 Heart 601
17.4.5 Kidney 602
17.4.6 Prostate 602
17.4.7 Lung 602
17.4.8 Urinary Bladder 603
17.5 FF-OCT to Identify Spermatogenesis in Rat Testis 605
17.6 FF-OCT for the Analysis of Human Lobectomy
Specimens 607
17.7 Potential Clinical Applications of FF-OCT 611
17.8 Current Limitations of FF-OCT and Possible
Solutions 612

18 FF-OCT Imaging: A Tool for Human Breast and Brain Tissue

Characterization 617
Osnath Assayag
18.1 Introduction 617
18.2 Breast Tissue Features Recognition and
Pathological Modifications 618
18.3 Distinction between Benign/Normal and Malignant
Tissue 622
18.3.1 Malignant Tissue Characterization 622
18.3.2 Benign Lesions Identification 624
18.4 Breast Tissue Classification Using FF-OCT 625
18.4.1 Diagnostic Accuracy of FF-OCT Images 627
18.5 FF-OCT Imaging of Healthy and Tumorous Human
Brain Parenchyma 627
18.5.1 Human Brain Parenchyma Morphological
Structure Recognition 629
18.5.2 Benign Lesions Identification 630
18.5.3 Malignant Brain Lesion Imaging 635
18.6 Discussion and Conclusion 636
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Contents xvii

19 Full-Field Optical Coherence Microscopy in Ophthalmology 641

G. Latour, K. Grieve, G. Georges, L. Siozade, M. Paques,
V. Borderie, L. Hoffart, and C. Deumié
19.1 Introduction 641
19.2 Cornea 643
19.2.1 Morphology and Characterization 643
19.2.1.1 Description of the tissue 643
19.2.1.2 Conventional corneal imaging
techniques 646
19.2.2 FF-OCM Imaging of the Healthy Cornea 648
19.2.3 Pathological Corneas 651
19.2.3.1 Monitoring the evolution of
edema 651
19.2.3.2 Imaging of common pathologies 658
19.2.4 Characterization of Laser Ablations 664
19.2.5 Perspectives 667
19.2.5.1 Eye banking 667
19.2.5.2 Characterization of artificial
corneas 669
19.3 Lens 669
19.3.1 Anatomy 669
19.3.2 Imaging the Lens 669
19.3.3 FF-OCM Lens Imaging 670
19.3.4 Conclusion and Perspectives 670
19.4 Retina 671
19.4.1 Anatomy 671
19.4.2 Retinal Imaging Techniques 672
19.4.3 FF-OCM Retinal Imaging 673
19.4.4 Perspectives in Retinal Imaging 675
19.5 Future Developments in Ophthalmology 677
19.5.1 In vivo FF-OCM Imaging 677
19.5.2 Coupling with Other Modalities: A Way to
Increase Information 677

20 Investigation of Spindle Structure and Embryo

Development for Preimplantation Genetic Diagnosis by
Subcellular Live Imaging with FF-OCT 689
Ping Xue and Jing-gao Zheng
20.1 Introduction 689
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xviii Contents

20.2 Experimental Design 691

20.3 Extraction of the Signals 693
20.3.1 Phase Shifting 693
20.3.2 Extract the Signals with Two-Phase Shifting 694
20.3.3 Comparison between Two and Four-Phase
Shifting 696
20.3.4 3D Reconstruction Image Display 697
20.4 System Characteristics 698
20.4.1 Axial Resolution 698
20.4.2 Transverse Resolution 699
20.4.3 Depth of Focus/Field 700
20.4.4 Sensitivity 701
20.5 Applications in Embryology and Developmental
Biology 704
20.5.1 Imaging the Structures of Spindles 705
20.5.2 Imaging Early Patterning and Polarity 710
20.5.2.1 Static studies of early patterning
and polarity 711
20.5.2.2 Dynamic studies of early
patterning and polarity 716
20.6 Conclusion 720

21 FF-OCT for Nondestructive Material Characterization and

Evaluation 727
David Stifter
21.1 Introduction 727
21.2 Methods 729
21.3 Applications 734
21.3.1 Surface Metrology 734
21.3.2 Layer Thickness Determination 737
21.3.3 3D Structural and Functional Evaluation 738
21.4 Conclusions and Outlook 745

Index 755
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Preface

Microscopic imaging for the visualization of the internal structure

of objects, nondestructively, has been a subject of active research
and developments for applications in the fields of medicine, biology,
and materials science. In particular, the capability to noninvasively
explore the microstructure of biological tissues with a spatial
resolution similar to that of histology has prompted significant work
in recent years.
Confocal microscopy is an optical imaging method to generate
high-resolution (≤1 μm) cross-sectional views of semitransparent
objects by rejecting light coming from out-of-focus regions. The
imaging penetration depth of confocal microscopy in most biological
tissues, however, is relatively weak (∼200 μm). Using ultrashort
pulse lasers, nonlinear optical effects such as harmonic generation
and multiphoton absorption have been successfully applied in
optical microscopy, offering deeper imaging penetration in highly
scattering tissues compared with confocal microscopy. Optical
coherence tomography (OCT) is another technique for noninvasive
imaging in semitransparent objects. OCT allows deeper imaging
(∼1 mm), but with a lower spatial resolution. In OCT, light
backscattered within the object is detected using low-coherence
interferometry. Cross-sectional images are generated by performing
axial measurements (A-scans) at different transverse positions.
OCT has revolutionized the clinical practice of ophthalmology for
the diagnosis and management of retinal diseases. OCT has been
clinically demonstrated in a diverse set of other medical and
surgical applications, including cardiology, gastroenterology, and
dermatology, allowing the visualization of the microstructure of
tissues without excision and processing.
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xx Preface

Full-field OCT (FF-OCT), also termed full-field optical coherence

microscopy (FF-OCM), is a particular version of OCT that uses
full-field illumination and an array detector to acquire en face
tomographic images without the need for transverse scanning. FF-
OCM benefits from the transverse imaging resolution of optical
microscopy along with the capacity of optical axial sectioning at
micrometer-scale resolution. Since the introduction of FF-OCM in
the early 2000s, significant progress in the technology has been
achieved. FF-OCM has been demonstrated in a variety of applications
and is now commercially available.
The diversity of scientific publications on FF-OCM has led to a
need for a comprehensive handbook describing FF-OCM in terms of
technology, performance, and applications. The aim of this book is to
address this need by serving as a guide for engineers and scientists
involved in the technological developments of FF-OCM. This book
may also constitute a reference for users of FF-OCM who wish to
understand the fundamentals of this imaging modality and to know
its capabilities and limitations.
This handbook comprises 21 self-contained chapters, organized
into four parts, written by internationally recognized experts and
leaders in the field of FF-OCM. The first chapter provides a general
introduction to FF-OCM. The fundamental characteristics of the
technology are analyzed and discussed theoretically in the following
four chapters, constituting the first part of the book. The recent
main technological developments of FF-OCM, in particular for
increasing the image acquisition speed, are presented in the second
part. Extensions of FF-OCM for image contrast enhancement and
functional imaging are reported in the third part. An overview of
applications of FF-OCM in medicine, biology, and materials science
is provided in the last part.
This book would not have been possible without the invaluable
contribution of many people. I would like to express my sincere
appreciation to all of my colleagues who have dedicated many hours
to writing the chapters. I am grateful to Stanford Chong, director and
publisher at Pan Stanford Publishing, for having initiated this work.
I also thank Sarabjeet Garcha for the editing work.
May 13, 2016 10:35 PSP Book - 9in x 6in 00-Arnaud-Dubois-Prelims

Preface xxi

I and all the coauthors hope you will find this handbook
interesting, stimulating, and useful.

Arnaud Dubois
Palaiseau, France
May 2016
May 12, 2016 16:15 PSP Book - 9in x 6in 01-Arnaud-Dubois-c01

Chapter 1

Introduction to Full-Field Optical

Coherence Microscopy

Arnaud Dubois
Laboratoire Charles Fabry, CNRS, Institut d’Optique Graduate School,
Univ. Paris-Saclay, 2 av. Augustin Fresnel, 91127 Palaiseau, France
arnaud.dubois@institutoptique.fr

This introductory chapter provides an overview of full-field optical

coherence microscopy. The different technological approaches that
have been developed are reported, specifying the interferometer
configuration, the light source, the image sensor, and the method
used to obtain images from the acquired data. Theoretical analyses
of the system characteristics, including the detection sensitivity
and spatial resolution, are presented. Challenges, advantages, and
drawbacks of the technique are discussed. A review of the main
applications of full-field optical coherence microscopy is provided.

1.1 Introduction

Optical microscopy has been a standard tool in life sciences as

well as material sciences for over the last century and a half.

Handbook of Full-Field Optical Coherence Microscopy: Technology and Applications

Edited by Arnaud Dubois
Copyright c 2016 Pan Stanford Publishing Pte. Ltd.
ISBN 978-981-4669-16-0 (Hardcover), 978-981-4669-17-7 (eBook)
www.panstanford.com
May 12, 2016 16:15 PSP Book - 9in x 6in 01-Arnaud-Dubois-c01

2 Introduction to Full-Field Optical Coherence Microscopy

Features of an object that are normally not visible to the naked

eye can be accessed using an optical microscope by increasing the
spatial resolution and providing image contrast. Three-dimensional
microscopic imaging for the visualization of the internal microstruc-
tures of semitransparent objects, mainly in biomedical applications,
has prompted intense technological developments. Diagnosis and
study of many diseases, especially cancers, depend heavily upon
biopsy and histopathological analysis of tissue at the cellular-
level scale using an optical microscope. A large variety of stains
and fluorescent dyes can be employed to enhance the contrast
of the image and differentiate individual components within the
tissue. However, as the penetration depth of visible light is very
limited, only thin and transparent samples can be visualized with
conventional optical microscopy. The imaged sample must be cut
into slices a few micrometers thick after being frozen or embedded
in paraffin, which is time consuming and may introduce structural
deformation or damage. Moreover, some biological components
can be deteriorated or removed during tissue processing. There
is therefore great interest for an optical technique capable of
noninvasive optical biopsy, that is, capable of real-time in situ
imaging of tissue microstructures with a resolution similar to that
of histology without the need for tissue excision and processing, or
at least for a technique capable of immediate tomographic imaging
of excised specimens at high resolution, while avoiding the long and
delicate histological preparations.
The introduction of confocal microscopy was a significant
advance in optical microscopy. By rejecting light coming from out-
of-focus regions, optical sections can be produced using a confocal
microscope without requiring physical sectioning. By labeling with
fluorophores, specimens can be visualized in three dimensions at
high resolution. However, the imaging penetration depth of confocal
microscopy is limited to ∼200 μm due to tissue scattering. Besides,
the use of fluorophores may affect the viability of cells.
With the great advances in the technology of ultrashort pulsed
lasers, nonlinear optical effects such as harmonic generation and
two-photon absorption have been applied successfully in the field
of microscopy. The advantage of two-photon excitation fluorescence
microscopy over linear (one-photon) fluorescence microscopy
May 12, 2016 16:15 PSP Book - 9in x 6in 01-Arnaud-Dubois-c01

Introduction 3

Figure 1.1 Comparison of typical resolution and imaging depth achieved

with different optical imaging techniques.

includes efficient background rejection, reduced photobleaching

and photodamage [1], and deeper imaging penetration because of
a longer excitation wavelength. However, the penetration is limited
to several hundreds of micrometers in highly scattering tissues [2].
Optical coherence tomography (OCT) is another technique for
three-dimensional imaging that can penetrate deeper than confocal
microscopy and nonlinear microscopy (see Fig. 1.1). OCT relies
on low-coherence interferometry to measure the amplitude of
light backscattered by the sample being imaged [3–6]. The most
significant impact of OCT is in ophthalmology for in situ examination
of the pathologic changes of the retina [7–10] and measurement
of the dimensions of the anterior chamber [11, 12]. OCT has also
been applied successfully to imaging of various highly scattering
tissues [13, 14]. Ultrahigh axial resolution of ∼1 μm can be achieved
by using laser-based ultrabroad-bandwidth light sources [15–17].
However, OCT usually suffers from a limited transverse (lateral)
resolution because relatively low-numerical-aperture lenses have
to be used to preserve a sufficient depth of field. Several solutions
have been implemented to improve the transverse resolution of OCT,
including adjustment of focus while the depth is scanned [15–18],
illumination of the sample with a Bessel beam [19, 20], or image
postprocessing based on inverse scattering theory [21]. The most
efficient approach for achieving fine enough transverse resolution
for cellular-level imaging in scattering tissues is to acquire en
face (parallel to the sample surface) images rather than cross-
sectional (perpendicular to the sample surface) images. In this
May 12, 2016 16:15 PSP Book - 9in x 6in 01-Arnaud-Dubois-c01

4 Introduction to Full-Field Optical Coherence Microscopy

configuration, there is no limitation to the depth of field. Optical

coherence microscopy, or OCM, is a version of OCT that produces
en face tomographic images. High-numerical-aperture optics can be
used in OCM to achieve higher transverse resolution. Two general
approaches for OCM have been reported to date. The first approach
is based on the combination of confocal microscopy with low-
coherence interferometry, as demonstrated in Refs. [22, 23]. This
combination was revisited later using modern technologies, leading
to the scanning OCM technique [24, 25]. Broadband coherence
gating was shown to significantly enhance the imaging depth of
conventional confocal microscopy [24, 26]. Scanning OCM was
applied successfully to cellular-level resolution imaging deep below
the surface of various human tissues such as skin [27], oral mucosa
[28], and colonic mucosa [26, 29, 30]. The second approach of
OCM involves full-field illumination and detection. Also sometimes
termed “full-field optical coherence tomography” (FF-OCT), full-
field optical coherence microscopy (FF-OCM) is an alternative
technique to scanning OCM, based on low-coherence (white-light)
interference microscopy [31–34]. FF-OCM produces tomographic
images in the en face orientation by arithmetic combination of
several interferometric images acquired with an area camera and by
illuminating the whole field to be imaged with low-coherence light.
The major interest for FF-OCM lies in its high imaging resolution
in both transverse and axial directions using a simple and robust
experimental arrangement [35, 36].
This chapter gives an overview of FF-OCM. The key technological
elements of the technology are discussed, showing their influence
on the system performance. A model of the detection sensitivity
and spatial resolution of FF-OCM is presented. Finally, the main
applications of FF-OCM are described.

1.2 Experimental Setup

1.2.1 General Principle

FF-OCM is based on the combination of a microscope with
a Michelson-type interferometer. The conventional experimental
May 12, 2016 16:15 PSP Book - 9in x 6in 01-Arnaud-Dubois-c01

Experimental Setup 5

Figure 1.2 Diagram of conventional FF-OCM. MO: microscope objective;

PZT: piezoelectric transducer.

setup is represented schematically in Fig. 1.2. The beam emitted by

the broadband light source is split into two beams by a beam splitter,
one of these beams being directed onto the sample and the other
onto a flat low-reflectivity reference mirror. Light backscattered
from the sample is combined at the beam splitter with light reflected
by the reference mirror. The resulting interferometric image is
formed on the image sensor of an area camera. A phase modulation
is introduced in the interferometer, usually by varying the position
of the reference mirror using a piezoelectric actuator. An arithmetic
combination of several phase-shifted interferometric images yields
an en face tomographic image of the sample.

1.2.2 Interference Microscope Configuration

Several configurations of interference microscopes have been used
in FF-OCM, referred to as Michelson, Mirau, and Linnik interference
microscopes (see Fig. 1.3).
May 12, 2016 16:15 PSP Book - 9in x 6in 01-Arnaud-Dubois-c01

6 Introduction to Full-Field Optical Coherence Microscopy

Figure 1.3 Interference microscope configurations used in FF-OCM. MO:

microscope objective; RM: reference mirror; S: sample; BS: beam splitter.

In the Michelson interference microscope, a single microscope

objective images both the sample and the reference mirror. The
Mirau interferometer is a variant consisting of a beam-splitting
surface and a reference mirror, arranged in such a way that both
optical elements share the same axis with the microscope objective.
The compactness of the Michelson and Mirau interferometers allows
for incorporation into the microscope objective. The presence of the
beam splitter and the reference mirror between the objective and
the sample requires a relatively long working distance and prevents
the use of a high numerical aperture.
Alternatively, the Linnik configuration requires two identical
microscope objectives, one placed in each arm on the interferometer.
Compared to the Michelson and Mirau interference microscopes,
the Linnik configuration is less compact but offers more flexibility
in adjustments. The optical length of the interferometer arms and
the focus can be adjusted independently. Moreover, immersion
objectives can be used, which is more difficult in the Michelson and
Mirau configurations, although this has been demonstrated [37–39].
There is no limitation in the numerical aperture of the objectives of
the Linnik microscope, which makes this configuration preferable
when high lateral imaging resolution is desired [31, 35].

1.2.3 Light Source

The light source is a key element in FF-OCM. Various features of the
light source, including spectral properties, noise, power, and spatial
Another random document with
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buttons here for his little boy at home, and gave them
to me to deliver, as he was about to die. Have them
sewed on to my pants for safe keeping.
July 14.—We have been too busy with the raiders
of late to manufacture any exchange news, and now
all hands are at work trying to see who can tell the
biggest yarns. The weak are feeling well to-night over
the story that we are all to be sent North this month,
before the 20th. Have not learned that the news
came from any reliable source. Rumors of
midsummer battles with Union troops victorious. It’s
“bite dog, bite bear,” with most of us prisoners; we
don’t care which licks, what we want is to get out of
this pen. Of course, we all care and want our side to
win, but it’s tough on patriotism. A court is now held
every day and offenders punished, principally by
buck and gagging, for misdemeanors. The hanging
has done worlds of good, still there is much stealing
going on yet, but in a sly way, not openly. Hold my
own as regards health. The dreaded month of July is
half gone, almost, and a good many over one
hundred and fifty die each day, but I do not know how
many. Hardly any one cares enough about it to help
me any in my inquiries. It is all self with the most of
them. A guard by accident shot himself. Have often
said they didn’t know enough to hold a gun. Bury a
rebel guard every few days within sight of the prison.
Saw some women in the distance. Quite a sight. Are
feeling quite jolly to-night since the sun went down.
Was visited by my new acquaintances of the 9th
Michigan Infantry, who are comparatively new
prisoners. Am learning them the way to live here.
They are very hopeful fellows and declare the war
will be over this coming fall, and tell their reasons
very well for thinking so. We gird up our loins and
decide that we will try to live it through. Rowe,
although often given to despondency, is feeling good
and cheerful. There are some noble fellows here. A
man shows exactly what he is in Andersonville. No
occasion to be any different from what you really are.
Very often see a great big fellow in size, in reality a
baby in action, actually sniveling and crying, and then
again you will see some little runt, “not bigger than a
pint of cider,” tell the big fellow to “brace up” and be a
man. Stature has nothing to do as regards nerve, still
there are noble big fellows as well as noble little
ones. A Sergt. Hill is judge and jury now, and
dispenses justice to evil doers with impartiality. A
farce is made of defending some of the arrested
ones. Hill inquires all of the particulars of each case,
and sometimes lets the offenders go as more sinned
against than sinning. Four receiving punishment.
July 15.—Blank cartridges were this morning fired
over the camp by the artillery, and immediately the
greatest commotion outside. It seems that the signal
in case a break is made, is cannon firing. And this
was to show us how quick they could rally and get
into shape. In less time than it takes for me to write it,
all were at their posts and in condition to open up
and kill nine-tenths of all here. Sweltering hot. Dying
off one hundred and fifty-five each day. There are
twenty-eight thousand confined here now.
July 16.—Well, who ever supposed that it could be
any hotter; but to-day is more so than yesterday, and
yesterday more than the day before. My coverlid has
been rained on so much and burned in the sun, first
one and then the other, that it is getting the worse for
wear. It was originally a very nice one, and home
made. Sun goes right through it now, and reaches
down for us. Just like a bake oven. The rabbit mules
that draw in the rations look as if they didn’t get much
more to eat than we do. Driven with one rope line,
and harness patched up with ropes, strings, &c. Fit
representation of the Confederacy. Not much like U.
S. Army teams. A joke on the rebel adjutant has
happened. Some one broke into the shanty and tied
the two or three sleeping there, and carried off all the
goods. Tennessee Bill, (a fellow captured with me)
had charge of the affair, and is in disgrace with the
adjutant on account of it. Every one is glad of the
robbery. Probably there was not ten dollars worth of
things in there, but they asked outrageous prices for
everything. Adjt. very mad, but no good. Is a small,
sputtering sort of fellow.
July 17.—Cords contracting in my legs and very
difficult for me to walk—after going a little ways have
to stop and rest and am faint. Am urged by some to
go to the hospital but don’t like to do it; mess say had
better stay where I am, and Battese says shall not
go, and that settles it. Jimmy Devers anxious to be
taken to the hospital but is pursuaded to give it up.
Tom McGill, another Irish friend, is past all recovery;
is in another part of the prison. Many old prisoners
are dropping off now this fearful hot weather; knew
that July and August would thin us out; cannot keep
track of them in my disabled condition. A fellow
named Hubbard with whom I have conversed a good
deal, is dead; a few days ago was in very good
health, and its only a question of a few days now with
any of us. Succeeded in getting four small onions
about as large as hickory nuts, tops and all for two
dollars Confederate money. Battese furnished the
money but won’t eat an onion; ask him if he is afraid
it will make his breath smell? It is said that two or
three onions or a sweet potato eaten raw daily will
cure the scurvy. What a shame that such things are
denied us, being so plenty the world over. Never
appreciated such things before but shall hereafter.
Am talking as if I expected to get home again. I do.
July 18.—Time slowly dragging itself along. Cut
some wretchs hair most every day. Have a sign out
“Hair Cutting,” as well as “Washing,” and by the way,
Battese has a new wash board made from a piece of
the scaffold lumber. About half the time do the work
for nothing, in fact not more than one in three or four
pays anything—expenses not much though, don’t
have to pay any rent. All the mess keeps their hair
cut short which is a very good advertisement. My
eyes getting weak with other troubles. Can just
hobble around. Death rate more than ever, reported
one hundred and sixty-five per day; said by some to
be more than that, but 165 is about the figure. Bad
enough without making any worse than it really is.
Jimmy Devers most dead and begs us to take him to
the hospital and guess will have to. Every morning
the sick are carried to the gate in blankets and on
stretchers, and the worst cases admitted to the
hospital. Probably out of five or six hundred half are
admitted. Do not think any lives after being taken
there; are past all human aid. Four out of every five
prefer to stay inside and die with their friends rather
than go to the hospital. Hard stories reach us of the
treatment of the sick out there and I am sorry to say
the cruelty emanates from our own men who act as
nurses. These dead beats and bummer nurses are
the same bounty jumpers the U. S. authorities have
had so much trouble with. Do not mean to say that all
the nurses are of that class but a great many of them
are.
July 19.—There is no such thing as delicacy here.
Nine out of ten would as soon eat with a corpse for a
table as any other way. In the middle of last night I
was awakened by being kicked by a dying man. He
was soon dead. In his struggles he had floundered
clear into our bed. Got up and moved the body off a
few feet, and again went to sleep to dream of the
hideous sights. I can never get used to it as some do.
Often wake most scared to death, and shuddering
from head to foot. Almost dread to go to sleep on this
account. I am getting worse and worse, and prison
ditto.
July 20.—Am troubled with poor sight together with
scurvy and dropsy. My teeth are all loose and it is
with difficulty I can eat. Jimmy Devers was taken out
to die to-day. I hear that McGill is also dead. John
McGuire died last night, both were Jackson men and
old acquaintances. Mike Hoare is still policeman and
is sorry for me. Does what he can. And so we have
seen the last of Jimmy. A prisoner of war one year
and eighteen days. Struggled hard to live through it,
if ever any one did. Ever since I can remember have
known him. John Maguire also, I have always known.
Everybody in Jackson, Mich., will remember him, as
living on the east side of the river near the
wintergreen patch, and his father before him. They
were one of the first families who settled that country.
His people are well to do, with much property.
Leaves a wife and one boy. Tom McGill is also a
Jackson boy and a member of my own company.
Thus you will see that three of my acquaintances
died the same day, for Jimmy cannot live until night I
don’t think. Not a person in the world but would have
thought either one of them would kill me a dozen
times enduring hardships. Pretty hard to tell about
such things. Small squad of poor deluded Yanks
turned inside with us, captured at Petersburg. It is
said they talk of winning recent battles. Battese has
traded for an old watch and Mike will try to procure
vegetables for it from the guard. That is what will
save us if anything.
July 21.—And rebels are still fortifying. Battese has
his hands full. Takes care of me like a father. Hear
that Kilpatrick is making a raid for this place. Troops
(rebel) are arriving here by every train to defend it.
Nothing but corn bread issued now and I cannot eat
it any more.
July 22.—A petition is gotten up signed by all the
sergeants in the prison, to be sent to Washington, D.
C., begging to be released. Capt. Wirtz has
consented to let three representatives go for that
purpose. Rough that it should be necessary for us to
beg to be protected by our government.
July 23.—Reports of an exchange in August. Can’t
stand it till that time. Will soon go up the spout.
July 24.—Have been trying to get into the hospital,
but Battese won’t let me go. Geo. W. Hutchins,
brother of Charlie Hutchins of Jackson, Mich., died
to-day—from our mess. Jimmy Devers is dead.
July 25.—Rowe getting very bad. Sanders ditto.
Am myself much worse, and cannot walk, and with
difficulty stand up. Legs drawn up like a triangle,
mouth in terrible shape, and dropsy worse than all. A
few more days. At my earnest solicitation was carried
to the gate this morning, to be admitted to the
hospital. Lay in the sun for some hours to be
examined, and finally my turn came and I tried to
stand up, but was so excited I fainted away. When I
came to myself I lay along with the row of dead on
the outside. Raised up and asked a rebel for a drink
of water, and he said: “Here, you Yank, if you ain’t
dead, get inside there!” And with his help was put
inside again. Told a man to go to our mess and tell
them to come to the gate, and pretty soon Battese
and Sanders came and carried me back to our
quarters; and here I am, completely played out.
Battese flying around to buy me something good to
eat. Can’t write much more. Exchange rumors.
July 26.—Ain’t dead yet. Actually laugh when I
think of the rebel who thought if I wasn’t dead I had
better get inside. Can’t walk a step now. Shall try for
the hospital no more. Had an onion.
July 27.—Sweltering hot. No worse than yesterday.
Said that two hundred die now each day. Rowe very
bad and Sanders getting so. Swan dead, Gordon
dead, Jack Withers dead, Scotty dead, a large
Irishman who has been near us a long time is dead.
These and scores of others died yesterday and day
before. Hub Dakin came to see me and brought an
onion. He is just able to crawl around himself.
July 28.—Taken a step forward toward the
trenches since yesterday, and am worse. Had a
wash all over this morning. Battese took me to the
creek; carries me without any trouble.
July 29.—Alive and kicking. Drank some soured
water made from meal and water.
July 30.—Hang on well, and no worse.
 MOVED JUST IN TIME.

REMOVED FROM ANDERSONVILLE TO THE MARINE HOSPITAL,

SAVANNAH—GETTING THROUGH THE GATE—BATTESE HAS
SAVED US—VERY SICK BUT BY NO MEANS DEAD YET—
BETTER AND HUMANE TREATMENT.

Aug. 1.—Just about the same. My Indian friend

says: “We all get away.”
Aug. 2.—Two hundred and twenty die each day.
No more news of exchange.
Aug. 3.—Had some good soup, and feel better. All
is done for me that can be done by my friends. Rowe
and Sanders in almost as bad a condition as myself.
Just about where I was two or three weeks ago.
Seem to have come down all at once. August goes
for them.
Aug. 4.—Storm threatened. Will cool the
atmosphere. Hard work to write.
Aug. 5.—Severe storm. Could die in two hours if I
wanted to, but don’t.
 Aug. 12.—Warm. Warm. Warm. If I only had some
shade to lay in, and a glass of lemonade.
Aug. 13.—A nice spring of cold water has broken
out in camp, enough to furnish nearly all here with
drinking water. God has not forgotten us. Battese
brings it to me to drink.
Aug. 14.—Battese very hopeful, as exchange
rumors are afloat. Talks more about it than ever
before.
Aug. 15.—The water is a God-send. Sanders
better and Rowe worse.
Aug. 16.—Still in the land of the living. Capt. Wirtz
is sick and a Lieut. Davis acting in his stead.
Aug. 17.—Hanging on yet. A good many more
than two hundred and twenty-five die now in twenty-
four hours. Messes that have stopped near us are all
dead.
Aug. 18.—Exchange rumors.
Aug. 19.—Am still hoping for relief. Water is
bracing some up, myself with others. Does not hurt
us.
Aug. 20.—Some say three hundred now die each
day. No more new men coming. Reported that Wirtz
is dead.
 Aug. 21.—Sleep nearly all the time except when
too hot to do so.
Aug. 22—Exchange rumors.
Aug. 23.—Terribly hot.
Aug. 24.—Had some soup. Not particularly worse,
but Rowe is, and Sanders also.
Aug. 25.—In my exuberance of joy must write a
few lines. Received a letter from my brother, George
W. Ransom, from Hilton Head.[A] Contained only a
few words.
[A] My brother supposed me dead, as I
had been so reported; still, thinking it
might not be so, every week or so he
would write a letter and direct to me as a
prisoner of war. This letter, very strangely,
reached its destination.
Aug. 26.—Still am writing. The letter from my
brother has done good and cheered me up. Eye sight
very poor and writing tires me. Battese sticks by;
such disinterested friendship is rare. Prison at its
worst.
Aug. 27.—Have now written nearly through three
large books, and still at it. The diary am confident will
reach my people if I don’t. There are many here who
are interested and will see that it goes north.
Aug. 28.—No news and no worse; set up part of
the time. Dying off a third faster than ever before.
Aug. 29.—Exchange rumors afloat. Any kind of a
change would help me.
Aug. 30.—Am in no pain whatever, and no worse.
Aug. 31.—Still waiting for something to turn up. My
Indian friend says: “good news yet.” Night.—The
camp is full of exchange rumors.
Sept 1.—Sanders taken outside to butcher cattle.
Is sick but goes all the same. Mike sick and no longer
a policeman. Still rumors of exchange.
Sept. 2.—Just about the same; rumors afloat does
me good. Am the most hopeful chap on record.
Sept. 3.—Trade off my rations for some little luxury
and manage to get up quite a soup. Later.—
Sanders sent in to us a quite large piece of fresh
beef and a little salt; another God-send.
Sept. 4.—Anything good to eat lifts me right up,
and the beef soup has done it.
Sept. 4.—The beef critter is a noble animal. Very
decided exchange rumors.
 Sept. 5.—The nice spring of cold water still flows
and furnishes drinking water for all; police guard it
night and day so to be taken away only in small
quantities. Three hundred said to be dying off each
day.
Sept. 6.—Hurrah! Hurrah!! Hurrah!!! Can’t holler
except on paper. Good news. Seven detachments
ordered to be ready to go at a moment’s notice.
Later.—All who cannot walk must stay behind. If left
behind shall die in twenty-four hours. Battese says I
shall go. Later.—Seven detachments are going out
of the gate; all the sick are left behind. Ours is the
tenth detachment and will go to-morrow so said. The
greatest excitement; men wild with joy. Am worried
fearful that I cannot go, but Battese says I shall.
Sept. 7.—Anxiously waiting the expected
summons. Rebels say as soon as transportation
comes, and so a car whistle is music to our ears.
Hope is a good medicine and am sitting up and have
been trying to stand up but can’t do it; legs too
crooked and with every attempt get faint. Men laugh
at the idea of my going, as the rebels are very
particular not to let any sick go, still Battese say I am
going. Most Dark.—Rebels say we go during the
night when transportation comes. Battese grinned
when this news come and can’t get his face
straightened out again.
 Marine Hospital, Savannah, Ga., Sept. 15, 1864.
—A great change has taken place since I last wrote
in my diary. Am in heaven now compared with the
past. At about midnight, September 7th, our
detachment was ordered outside at Andersonville,
and Battese picked me up and carried me to the
gate. The men were being let outside in ranks of four,
and counted as they went out. They were very strict
about letting none go but the well ones, or those who
could walk. The rebel adjutant stood upon a box by
the gate, watching very close. Pitch pine knots were
burning in the near vicinity to give light. As it came
our turn to go Battese got me in the middle of the
rank, stood me up as well as I could stand, and with
himself on one side and Sergt. Rowe on the other
began pushing our way through the gate. Could not
help myself a particle, and was so faint that I hardly
knew what was going on. As we were going through
the gate the adjutant yells out: “Here, here! hold on
there, that man can’t go, hold on there!” and Battese
crowding right along outside. The adjutant struck
over the heads of the men and tried to stop us, but
my noble Indian friend kept straight ahead, hallooing:
“He all right, he well, he go!” And so I got outside,
and adjutant having too much to look after to follow
me. After we were outside, I was carried to the
railroad in the same coverlid which I fooled the rebel
out of when captured, and which I presume has
saved my life a dozen times. We were crowded very
thick into box cars. I was nearly dead, and hardly
knew where we were or what was going on. We were
two days in getting to Savannah. Arrived early in the
morning. The railroads here run in the middle of very
wide, handsome streets. We were unloaded, I should
judge, near the middle of the city. The men as they
were unloaded, fell into line and were marched away.
Battese got me out of the car, and laid me on the
pavement. They then obliged him to go with the rest,
leaving me; would not let him take me. I lay there
until noon with four or five others, without any guard.
Three or four times negro servants came to us from
houses near by, and gave us water, milk and food.
With much difficulty I could set up, but was
completely helpless. A little after noon a wagon came
and toted us to a temporary hospital in the outskirts
of the city, and near a prison pen they had just built
for the well ones. Where I was taken it was merely an
open piece of ground, having wall tents erected and
a line of guards around it. I was put into a tent and
lay on the coverlid. That night some gruel was given
to me, and a nurse whom I had seen in Andersonville
looked in, and my name was taken. The next
morning, September 10th, I woke up and went to
move my hands, and could not do it; could not move
either limb so much as an inch. Could move my head
with difficulty. Seemed to be paralyzed, but in no pain
whatever. After a few hours a physician came to my
tent, examined and gave me medicine, also left
medicine, and one of the nurses fed me some soup
or gruel. By night I could move my hands. Lay awake
considerable through the night thinking. Was happy
as a clam in high tide. Seemed so nice to be under a
nice clean tent, and there was such cool pure air.
The surroundings were so much better that I thought
now would be a good time to die, and I didn’t care
one way or the other. Next morning the doctor came,
and with him Sergt. Winn. Sergt. Winn I had had a
little acquaintance with at Andersonville. Doctor said I
was terribly reduced, but he thought I would improve.
Told them to wash me. A nurse came and washed
me, and Winn brought me a white cotton shirt, and
an old but clean pair of pants; my old clothing, which
was in rags, was taken away. Two or three times
during the day I had gruel of some kind, I don’t know
what. Medicine was given me by the nurses. By night
I could move my feet and legs a little. The cords in
my feet and legs were contracted so, of course, that I
couldn’t straighten myself out. Kept thinking to
myself, “am I really away from that place
Andersonville?” It seemed too good to be true. On
the morning of the 12th, ambulances moved all to the
Marine Hospital, or rather an orchard in same yard
with Marine Hospital, where thirty or forty nice new
tents have been put up, with bunks about two feet
from the ground, inside. Was put into a tent. By this
time could move my arms considerable. We were
given vinegar weakened with water, and also salt in
it. Had medicine. My legs began to get movable more
each day, also my arms, and to-day I am laying on
my stomach and writing in my diary. Mike Hoare is
also in this hospital. One of my tent mates is a man
named Land, who is a printer, same as myself. I hear
that Wm. B. Rowe is here also, but haven’t seen him.
Sept. 16.—How I do sleep; am tired out, and
seems to me I can just sleep till doomsday.
Sept. 17.—Four in each tent. A nurse raises me
up, sitting posture, and there I stay for hours, dozing
and talking away. Whiskey given us in very small
quantities, probably half a teaspoonful in half a glass
of something, I don’t know what. Actually makes me
drunk. I am in no pain whatever.
Sept. 18.—Surgeon examined me very thoroughly
to-day. Have some bad sores caused by laying down
so much; put something on them that makes them
ache. Sergt. Winn gave me a pair of socks.
Sept. 19.—A priest gave me some alum for my
sore mouth. Had a piece of sweet potato, but couldn’t
eat it. Fearfully weak. Soup is all I can eat, and don’t
always stay down.
Sept. 20.—Too cool for me. The priest said he
would come and see me often. Good man. My left
hand got bruised in some way and rebel done it up.
He is afraid gangrene will get in sore. Mike Hoare is
quite sick.
Sept. 21.—Don’t feel as well as I did some days
ago. Can’t eat; still can use my limbs and arms more.
Sept. 22.—Good many sick brought here.
Everybody is kind, rebels and all. Am now differently
sick than at any other time. Take lots of medicine, eat
nothing but gruel. Surgeons are very attentive. Man
died in my tent. Oh, if I was away by myself, I would
get well. Don’t want to see a sick man. That makes
me sick.
Sept. 23.—Shall write any way; have to watch
nurses and rebels or will lose my diary. Vinegar
reduced I drink and it is good; crave after acids and
salt. Mouth appears to be actually sorer than ever
before, but whether it is worse or not can’t say. Sergt.
Winn says the Doctor says that I must be very careful
if I want to get well. How in the old Harry can I be
careful? They are the ones that had better be careful
and give me the right medicine and food. Gruel made
out of a dish cloth to eat.