Quantitative phase Microscopy (QPM) has become an important imaging technique in biology for investigating cells and tissues. QPM is an optical interference or holographic microscopic technique in which an input light beam is divided into two beams and one passes through the object and the other acts as a reference beam. This book develops and describes the most advanced QPM techniques and computational imaging techniques using partially spatially coherent monochromatic light rather than highly coherent lasers. Using partially coherent light as a light source instead of a traditional laser has significantly improved QPM imaging results. Imaging techniques that will be discussed include speckle-free QPM both off-axis and common path interferometric configurations, structured illumination phase microscopy (SIPM), chip-based nanoscopy, machine learning, deep learning and artificial intelligence (AI) in phase microscopy and OCT and multi-spectral and hyper-spectral phase microscopy. Coherent-noise free QPM techniques described in this book lead to an order of magnitude improved spatial phase sensitivity, space-bandwidth product, and high temporal phase stability. The technique was utilized for sperm cells, macrophages, liver sinusoidal endothelial cells, cancer cells and red blood cells for precise QPM. We have demonstrated that partially spatially coherent monochromatic light is a most suitable source for high precision QPM. The text is highly useful for biologists, optical engineers, optical scientists, researchers working in QPM and OCT, and graduate and post graduate students. Part of IOP Series in Advances in Optics, Photonics and Optoelectronics.
Author(s): Dalip Singh Mehta, Ankit Butola, Veena Singh
Publisher: IOP Publishing
Year: 2022
Language: English
Pages: 172
City: Bristol
PRELIMS.pdf
Preface
Acknowledgement
Author biographies
D S Mehta
Ankit Butola
Veena Singh
CH001.pdf
Chapter 1 Introduction
1.1 Bright field microscopy
1.1.1 Lateral (transverse) resolution
1.1.2 Axial (longitudinal) resolution
1.2 Phase contrast microscopy
1.3 Quantitative phase microscopy
1.4 Coherence properties of laser light
1.4.1 Temporal coherence properties
1.4.2 Spatial coherence properties of laser light
1.5 Origin of speckles in imaging and microscopy
1.6 Effects of speckle on imaging and microscopy
1.6.1 Space-bandwidth product of QPM
1.6.2 Effect of speckle on quantitative phase microscopy
1.7 Methods of generating partially spatially coherent light
References
CH002.pdf
Chapter 2 Partially spatially coherent off-axis quantitative phase microscopy
2.1 Off-axis holographic phase microscopy
2.1.1 Conventional (Leith and Upatnieks’) holography
2.1.2 Digital holography
2.2 Partially spatially coherent off-axis quantitative phase microscopy
2.2.1 Linnik interferometer based optical configuration
2.2.2 Mach–Zehnder interferometer based optical configuration
2.3 Image characteristics
2.3.1 Resolution
2.3.2 Signal-to-noise ratio
2.4 Extraction of quantitative information using phase map
References
CH003.pdf
Chapter 3 Partially spatially coherent common-path quantitative phase microscopy
3.1 Gabor (in-line) holography
3.2 Diffraction phase microscopy
3.3 Fourier phase microscopy
3.4 Lateral shearing interferometric phase microscopy
3.5 Fresnel bi-prism based interferometric phase microscopy
3.6 Spatial phase sensitivity
3.7 Temporal phase stability
3.8 Resolution of the system
References
CH004.pdf
Chapter 4 Structured illumination quantitative phase microscopy
4.1 Diffraction-limited resolution of phase microscopy
4.2 Structured illumination schemes for phase microscopy
4.2.1 Off-axis geometry for SIQPM
4.3 Common-path geometries for SIQPM
4.3.1 Structured illumination lateral shearing interferometry based SIQPM
4.3.2 Structured illumination using Fresnel bi-prism interferometer
4.3.3 Structured illumination diffraction phase microscopy
4.4 Phase reconstruction algorithms for SIPM
References
CH005.pdf
Chapter 5 Multimodal on-chip nanoscopy and quantitative phase microscopy
5.1 Fluorescence imaging
5.1.1 Epi-fluorescence imaging
5.2 Total internal reflections fluorescence microscopy
5.3 Super-resolution fluorescence imaging
5.3.1 On-chip nanoscopy (single-molecule localization microscopy technique)
5.3.2 Working principle of on-chip nanoscopy
5.3.3 Chip preparation and staining protocol
5.4 Integrated on-chip nanoscopy and partially spatially coherent quantitative phase microscopy
5.5 Applications of integrated on-chip nanoscopy and quantitative phase microscopy
References
CH006.pdf
Chapter 6 Longitudinal spatial coherence gated tomography using partially spatially coherent monochromatic light
6.1 Introduction to time-domain and frequency-domain optical coherence tomography
6.1.1 Time-domain OCT
6.1.2 Frequency-domain (spectral-domain) optical coherence tomography
6.1.3 Swept-source optical coherence tomography (SS-OCT)
6.2 Axial-resolution in OCT systems
6.3 Dispersion effects in OCT
6.4 Concept of longitudinal spatial coherence
6.4.1 Analogy between temporal and spatial coherence
6.4.2 Longitudinal spatial coherence
6.5 Longitudinal spatial coherence gated topography and tomography.
References
CH007.pdf
Chapter 7 Low-coherence (white light) interference microscopy with colour fringe analysis
7.1 Introduction to low coherence interferometry
7.2 Phase-shifting white light interference microscopy
7.2.1 Hilbert transform method for phase extraction
7.2.2 Experimental details of white light interferometry
7.3 Color fringe analysis
7.4 Quantitative information about biological samples
7.4.1 Quantitative phase microscopy of heathy and anaemic blood samples
7.4.2 Quantitative phase imaging of normal and cancerous cells
7.4.3 Quantitative phase imaging of Escherichia coli bacteria
References
CH008.pdf
Chapter 8 Artificial intelligence: a computational tool to interpret quantitative phase imaging
8.1 Introduction
8.2 Machine learning to understand QPI/need of machine learning and deep learning in phase microscopy
8.2.1 Convolutional neural network architecture
8.2.2 Conventional versus deep learning approach for classification of QPI dataset
8.2.3 Phase retrieval and virtual staining in QPI
8.2.4 High resolution quantitative phase imaging using machine learning
8.3 Practical prescription for machine learning in QPI
8.4 Outlook
References
CH009.pdf
Chapter 9 Multi-spectral and hyper-spectral phase microscopy
9.1 Introduction
9.2 Multi-spectral phase microscopy
9.3 Hyper-spectral quantitative phase microscopy
9.4 Optical configurations of multi-spectral and hyper-spectral phase microscopy
9.5 Light sources for multi-spectral and hyper-spectral phase microscopy
9.6 Recording devices for multi-spectral and hyper-spectral phase microscopy
9.7 Algorithms for image reconstruction
9.7.1 Fourier transform method
9.7.2 Phase-shifting method
9.8 Applications of multi-spectral and hyper-spectral phase microscopy
References
CH010.pdf
Chapter 10 Conclusions and future prospects
References
