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In this chapter, we introduce digital holographic microscopy (DHM) as amarker-free method to determine the refractive index of single, spherical cellsin suspension. The refractive index is a conclusive measure in a biologicalcontext. Cell conditions, such as differentiation or infection, are known toyield significant changes in the refractive index. Furthermore, the refractiveindex of biological tissue determines the way it interacts with light. Besidesthe biological relevance of this interaction in the retina, a lot of methodsused in biology, including microscopy, rely on light-tissue or light-cellinteractions. Hence, determining the refractive index of cells using DHM isvaluable in many biological applications. This chapter covers the main topicswhich are important for the implementation of DHM: setup, sample preparationand analysis. First, the optical setup is described in detail including notesand suggestions for the implementation. Following that, a protocol for thesample and measurement preparation is explained. In the analysis section, analgorithm for the determination of the quantitative phase map is described.Subsequently, all intermediate steps for the calculation of the refractiveindex of suspended cells are presented, exploiting their spherical shape. Inthe last section, a discussion of possible extensions to the setup, furthermeasurement configurations and additional analysis methods are given.Throughout this chapter, we describe a simple, robust, and thus easilyreproducible implementation of DHM. The different possibilities for extensionsshow the diverse fields of application for this technique.

Refractive index measurements of single, spherical cells using digital holographic microscopy