Accurate single-shot quantitative phase imaging of biological specimens with telecentric digital holographic microscopy

“…28,30 Multicellular organisms have been previously studied using optical techniques, but have been limited to studying the superficial structure and motion of the organism due to limited resolution, or have required physical sectioning of the sample for improved resolution of smaller structures. [31][32][33][34] We propose using QPI as a label-free method for studying the arthropod external morphology and subcuticular structures. In this study, using both SLIM and the Hamamatsu Nanozoomer, we imaged a whole unstained springtail (Arthropod, Collembola) Ceratophysella denticulata mounted on a slide.…”

Section: Introductionmentioning

confidence: 99%

Quantitative phase imaging of arthropods

et al. 2015

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Abstract. Classification of arthropods is performed by characterization of fine features such as setae and cuticles. An unstained whole arthropod specimen mounted on a slide can be preserved for many decades, but is difficult to study since current methods require sample manipulation or tedious image processing. Spatial light interference microscopy (SLIM) is a quantitative phase imaging (QPI) technique that is an addon module to a commercial phase contrast microscope. We use SLIM to image a whole organism springtail Ceratophysella denticulata mounted on a slide. This is the first time, to our knowledge, that an entire organism has been imaged using QPI. We also demonstrate the ability of SLIM to image fine structures in addition to providing quantitative data that cannot be obtained by traditional bright field microscopy.

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“…However, our research group has recently demonstrated that the optical capture stage controls the accuracy of QPI-DHM. [5][6][7] Specifically, we showed that QPI-DHM operating in telecentric mode-producing a 2D image of a 3D object-provides very accurate diffraction-limited quantitative phase images with minimal post-processing. This accuracy can be very useful for the diagnosis of different diseases and for specimen identification.…”

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confidence: 99%

Digital holographic microscopy for diabetes screening

Doblas¹,

Martı́nez-Corral²,

Saavedra³

2016

SPIE Newsroom

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A digital holographic microscope operating in telecentric mode could be used to diagnose diabetes and evaluate long-term glycemic control in patients with diabetes.Transparent objects, such as live biological cells, are difficult to observe and quantify using traditional light microscopes owing to lack of contrast. However, recent advances in quantitative phase imaging (QPI) of transparent microscopic specimens using digital holographic microscopy (DHM) have facilitated observation of unstained living cells in ways that were previously impossible. This type of microscopy relies on the different optical densities of the specimen and surrounding medium, producing a difference in phase between the light passing through the specimen and the light passing through the medium, ultimately generating an interference pattern. In DHM, only a single recorded image is required to obtain the fringe pattern resulting from the interference between a reference wave and the image of the wavefront diffracted by the specimen. From the pattern, phase changes introduced by the specimen are easily retrieved to produce a quantitative phase map. These phase changes encode the information about the refractive index and thickness of the specimen and as a result, QPI-DHM can be used to analyze illnesses in which the refractive index and/or morphology of cells or tissues are distorted. [1][2][3][4] The application of QPI-DHM to medical diagnosis relies on the high accuracy of the phase map measurements. The optical capture and subsequent computational processing both have a significant role in this hybrid technique. However, our research group has recently demonstrated that the optical capture stage controls the accuracy of QPI-DHM. 5-7 Specifically, we showed that QPI-DHM operating in telecentric mode-producing a 2D image of a 3D object-provides very accurate diffraction-limited quantitative phase images with minimal post-processing. This accuracy can be very useful for the diagnosis of different diseases and for specimen identification. Here, we show how this type of microscopy can be used in the diagnosis of diabetes mellitus (DM), using a simple procedure that requires a small amount of blood.Telecentric DHM has a potential application as a technique for the screening of DM, 8 which is undoubtedly one of the most challenging health problems of this century. Assessment and control of hyperglycemic states is one of the main methods of minimizing the risk of developing the medical complications associated with DM. Currently, the gold standard method to evaluate hyperglycemia in patients with DM and obtain long-term glycemic control is the determination of glycated hemoglobin (HbA1c) concentration. This measurement is usually made by high-performance liquid chromatography (HPLC), Continued on next page

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Accurate single-shot quantitative phase imaging of biological specimens with telecentric digital holographic microscopy

Cited by 92 publications

References 45 publications

Study of spatial lateral resolution in off-axis digital holographic microscopy

Study of spatial lateral resolution in off-axis digital holographic microscopy

Quantitative phase imaging of arthropods

Digital holographic microscopy for diabetes screening

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