Combining the polarization-contrast and interference-contrast methods for three-dimensional visualization of anisotropic microobjects

Abstract

This paper presents a theoretical and experimental study of the possibilities of using a holographic method of polarization contrast to solve the problem of three-dimensional visualization of anisotropic microobjects. The experimental studies were carried out using a digital holographic interference microscope. It is shown that the polarization-contrast method makes it possible to obtain three-dimensional images of anisotropic microobjects with the maximum contrast and high quality, and this increases the sensitivity and resolution of the method by comparison with other optical methods. The possibilities of combining holographic methods of polarization contrast and interference contrast for the three-dimensional visualization of anisotropic microobjects have been studied for the first time.

Contrast-tuneable microscopy for single-shot real-time imaging

“…On the contrary, the Zernike phase contrast microscopy (ZPCM) [12] does not require staining, yet suffers from the halo effect [13], object size inaccuracy, and a narrow field of view. Recently, dramatic progress has taken place in staining-free imaging in the burgeoning field of quantitative phase-contrast microscopy [3,4,[14][15][16][17], such as by differential interference microscopy [18][19][20], digital holographic microscopy [5,[21][22][23][24], and holographic interference microscopy [25,26].…”

“…It is coherent irradiation by laser beam that can simplify the optical setup and phase plate design (a pinhole instead of zonal-shaped) while achieving image clarity and high resolution [15,16]. In contrast to single-shot imaging performed in an 'analogue' manner, quantitative phase imaging, implemented through a wide variety of techniques [5,14,15,[20][21][22][23][24][25][26], constructs an image from multiple shots recorded digitally at multiple phase differences. Note that these techniques focus on thin phase objects with an emphasis on computational image reconstruction to achieve image clarity [5,14,32,15,[20][21][22][23][24][25][26].…”

“…Furthermore, many of the quantitative phase imaging approaches proposed so far focus on lower-order scattered beams [5,14,15,[20][21][22][23][24][25][26]. However, incorporating higher-order scattered beams is beneficial for image edge clarity and for observing microstructures smaller in scale than wavelength.…”

Phase and Polarization Contrast Methods by Use of Digital Holographic Microscopy: Applications to Different Types of Biological Samples

“…An associated technique which uses the phase information for studying the state of polarization of live neurons in culture was developed by (Wang et al, 2008). However, according to recent publications, the Holographic Microscopy for polarization imaging (Polarization Holographic Microscopy, PHM), has been poorly applied to biological specimens analysis but only to fiber optics (Colomb et al, 2005), polymers ) and other inorganic materials (Tishko et al, 2012). This demonstrates the need of studies using this method for new applications viewing the biomedical field.…”

Técnicas de microscopia holográfica digital aplicadas à análise de sistemas biológicos

“…Uma técnica associada, a qual utiliza a informação de fase para o estudo do estado de polarização de neurônios vivos em cultura, foi desenvolvida por Wang [21]. De acordo com as publicações mais recentes, a Microscopia Holográfica de Polarização (MHP) tem sido pouco aplicada à análise de sistemas biológicos, mais frequentemente ao estudo de fibras ópticas [22], polímeros [19] e outros materiais inorgânicos [23]. Esse fato sugere a necessidade de aplicações desse método visando o campo biomédico.…”