Multi-dimensional Imaging

Chapter 9Digital Holographic Microscopy: A New Imaging Technique to Quantitatively Explore Cell Dynamics with Nanometer Sensitivity

Pierre Marquet^1,2 and Christian Depeursinge³

¹Centre de Neurosciences Psychiatriques, Centre Hospitalier Universitaire Vaudois, Département de Psychiatrie, Switzerland

²Brain Mind Institute, Institute of Microengineering, École Polytechnique Fédérale de Lausanne, Switzerland

³Institute of Microengineering, École Polytechnique Fédérale de Lausanne, Switzerland

9.1 Chapter Overview

In the first part of this chapter, we summarize how the new concept of digital optics, applied to the field of holographic microscopy, has allowed the development of a reliable and flexible digital holographic quantitative phase microscopy (DH-QPM) technique at the nanoscale, particularly suitable for cell imaging. In the second part, particular emphasis is placed on the original biological information provided by the quantitative phase signal. We present the most relevant DH-QPM applications in the field of cell biology, including automated cell counts, recognition, classification, three-dimensional tracking, and discrimination between physiological and pathophysiological states. In addition, we present how the phase signal can be used to specifically calculate some important biophysical cell parameters including dry mass, protein content and production, membrane fluctuations at the nanoscale, absolute volume, transmembrane water permeability, and how these different ...