Advantages of digital holographic microscopy for real-time full field absolute phase imaging

Tristan Colomb; Florian Charrière; Jonas Kühn; Pierre Marquet; Christian Depeursinge

doi:10.1117/12.763284

20 February 2008 Advantages of digital holographic microscopy for real-time full field absolute phase imaging

Tristan Colomb, Florian Charrière, Jonas Kühn, Pierre Marquet, Christian Depeursinge

Proceedings Volume 6861, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV; 686109 (2008) https://doi.org/10.1117/12.763284
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

Different interferometric techniques were developed last decade to obtain full field, quantitative, and absolute phase imaging, such as phase-shifting, Fourier phase microscopy, Hilbert phase microscopy or digital holographic microscopy (DHM). Although, these techniques are very similar, DHM combines several advantages. In contrast, to phase shifting, DHM is indeed capable of single-shot hologram recording allowing a real-time absolute phase imaging. On the other hand, unlike to Fourier phase or Hilbert phase microscopy, DHM does not require to record in focus images of the specimen on the digital detector (CCD or CMOS camera), because a numerical focalization adjustment can be performed by a numerical wavefront propagation. Consequently, the depth of view of high NA microscope objectives is numerically extended. For example, two different biological cells, floating at different depths in a liquid, can be focalized numerically from the same digital hologram. Moreover, the numerical propagation associated to digital optics and automatic fitting procedures, permits vibrations insensitive full- field phase imaging and the complete compensation for a priori any image distortion or/and phase aberrations introduced for example by imperfections of holders or perfusion chamber. Examples of real-time full-field phase images of biological cells have been demonstrated.

Citation Download Citation

Tristan Colomb, Florian Charrière, Jonas Kühn, Pierre Marquet, and Christian Depeursinge "Advantages of digital holographic microscopy for real-time full field absolute phase imaging", Proc. SPIE 6861, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV, 686109 (20 February 2008); https://doi.org/10.1117/12.763284

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