Simplified approach for quantitative digital holographic phase contrast imaging of living cells

Abstract

Many interferometry-based quantitative phase contrast imaging techniques require a separately generated coherent reference wave. This results in a low phase stability and the demand for a precise adjustment of the intensity ratio between object and reference wave. To overcome these problems, the performance of a Michelson interferometer approach for digital holographic microscopy was analyzed that avoids a separately generated reference wave by superposition of different image areas. It is shown that this simplified arrangement yields improved phase stability. Furthermore, results from time-lapse investigations on living pancreas tumor cells demonstrate the capability of the method for reliable quantitative phase contrast imaging.

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

“…Important to note is that the different setups come with slightly different benefits and limitations, making them inherently well suited to a variety of purposes: High speed acquisition for high temporal resolution of flow imaging [70], in situ imaging of growing adherent cells [21,37,56], live cell tomography [71,72], or incubator monitoring [30,73].…”

“…Typically, it consists of one or several reconstruction stages using Fourier Transforms and a focal selection stage, followed by one or more post-processing calculations where modular phase recordings are unwrapped and edge effects removed [4]. The risk of introducing image artefacts in the unwrapping stage is reduced if the sample phase shift is less than one wavelength, or it can be avoided completely by imaging at several wavelengths, thus also allowing for imaging thicker specimens [73,77,78]. All image reconstruction and manipulation stages can be performed after capture, as the complete three-dimensional information of the sample is stored in the interferometric recording.…”

Enhanced quantitative phase imaging in self-interference digital holographic microscopy using an electrically focus tunable lens

“…Series of amplitude and phase distributions are reconstructed by existing numerical procedures and subsequently averaged to achieve enhanced amplitude and quantitative phase images. The method is demonstrated and found particularly convenient for a recently reported self-interference DHM principle [25]. However, the concept may also be used with other experimental setups like for example Mach-Zehnder interferometer-based DHM implementations.…”

“…However, the concept may also be used with other experimental setups like for example Mach-Zehnder interferometer-based DHM implementations. Figure 1 shows the used experimental setup that is based on an inverted research microscope (Zeiss Axio observer A1, Carl Zeiss Micro Imaging GmbH, Goettingen, Germany) with an attached self-interference DHM unit as already presented with details in [25]. A frequencydoubled Nd:yttrium aluminium garnet (YAG) laser (Laser-Compact Co. Ldt, Moscow, Russia), emitting light at λ = 532 nm with a coherence length of l c ≈250 µm is utilized for sample illumination via a single mode optical fiber (SM).…”

One Type of Bio-Phase Microscopy Imaging Method Based on a Con Beam with Double Dove Prisms

“…However, most of imaging techniques are based on the interference principle of two optical waves, since they have the advantages of high detection sensitivity, fast detection rate and high detection accuracy. Now, different kinds of interference imaging systems are produced, such as: Mach-Zehnder (MZ) interferometer [3], phase contrast microscopy interferometer [4], Nomarshi differential interferometer [5], Michelson interferometer [6,7], Linnik interferometer [8,9] and Mirau interferometer [10]. In addition, according to the recording mode, interferometric phase microscopy techniques are mainly classified as two categorites: on-axis and off-axis.…”