Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement

Zuo, Chao; Sun, Jiasong; Feng, Shijie; Hu, Yan; Chen, Qian

doi:10.1016/j.optlaseng.2015.09.009

Cited by 24 publications

(12 citation statements)

References 18 publications

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“…More recently, a further remarkable extension of computational light microscopy was achieved by the applications of programmable light sources, e.g., wavelengthtunable lasers [74] and light-emitting diode (LED) array [75]] and tunable optical elements [e.g., electrically tunable lens (ETL) [76], liquid crystal display (LCD) [77], and digital mirror device (DMD) [78], complementing the digital processing with a matching optical modulation capability. Such devices not only provide additional flexibilities of active illumination and aperture control to realize multi-modal microscopy but result in a bunch of new computational light microscopy approaches.…”

Section: Phase Changesmentioning

confidence: 99%

Smart Computational Light Microscopes (SCLMs) of Smart Computational Imaging Laboratory (SCILab)

Fan

et al. 2021

Preprint

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Computational microscopy, as a subfield of computational imaging, combines optical manipulation and image algorithmic reconstruction to recover multi-dimensional microscopic images or information of micro-objects. In recent years, the revolution in light-emitting diodes (LEDs), low-cost consumer image sensors, modern digital computers, and smartphones provide fertile opportunities for the rapid development of computational microscopy. Consequently, diverse forms of computational microscopy have been invented, including digital holographic microscopy (DHM), transport of intensity equation (TIE), differential phase contrast (DPC) microscopy, lens-free on-chip holography, and Fourier ptychographic microscopy (FPM). These computational microscopy techniques not only provide high-resolution, label-free, quantitative phase imaging capability but also decipher new and advanced biomedical research and industrial applications. Nevertheless, most computational microscopy techniques are still at an early stage of "proof of concept" or "proof of prototype" (based on commercially available microscope platforms). Translating those concepts to stand-alone optical instruments for practical use is an essential step for the promotion and adoption of computational microscopy by the wider bio-medicine, industry, and education community. In this paper, we present four smart computational light microscopes (SCLMs) developed by our laboratory, i.e., smart computational imaging laboratory (SCILab) of Nanjing University of Science and Technology (NJUST), China. These microscopes are empowered by advanced computational microscopy techniques, including digital holography, TIE, DPC, lensless holography, and FPM, which not only enables multi-modal contrast-enhanced observations for unstained specimens, but also can recover their three-dimensional profiles quantitatively. We introduce their basic principles, hardware configurations, reconstruction algorithms, and software design, quantify their imaging performance, and illustrate their typical applications for cell analysis, medical diagnosis, and microlens characterization.

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Section: Phase Changesmentioning

confidence: 99%

Smart Computational Light Microscopes (SCLMs) of Smart Computational Imaging Laboratory (SCILab)

Fan

et al. 2021

Preprint

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“…The diffraction pattern resulting from the phase delay introduced by the sample can be observed in other defocused planes, but such an approach renders the image difficult to interpret [1]. Several specialized imaging techniques [1][2][3][4][5][6][7][8][9][10][11][12][13][14] are commonly employed in light microscopes in order to enhance image contrast and enable a direct visualization of subcellular features (as well as other types of samples that induce small phase delays) without staining, such as dark field [1,2], phase contrast [3][4][5], differential interference contrast [6][7][8] (DIC), fluorescence [9,10], and Rheinberg illumination [11][12][13][14]. By exploiting refraction, diffraction, interference, or fluorescence, these methods are applied mainly to make visible objects such as cells and other biological structures that are otherwise invisible.…”

Section: Introductionmentioning

confidence: 99%

“…Although not as popular as phase contrast or DIC, Rheinberg illumination [11][12][13][14] is one such technique that provides a form of optical staining. The approach was initially demonstrated by the British microscopist Julius Rheinberg to the Royal Microscopical Society and the Quekett Club (England) in 1861.…”

Section: Introductionmentioning

confidence: 99%

Label-free color staining of quantitative phase images of biological cells by simulated Rheinberg illumination

et al. 2019

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Modern microscopes are designed with functionalities that are tailored to enhance image contrast. Dark-field imaging, phase contrast, differential interference contrast, and other optical techniques enable biological cells and other phase-only objects to be visualized. Quantitative phase imaging refers to an emerging set of techniques that allow for the complex transmission function of the sample to be measured. With this quantitative phase image available, any optical technique can then be simulated; it is trivial to generate a phase contrast image or a differential interference contrast image. Rheinberg illumination, proposed almost a century ago, is an optical technique that applies color contrast to images of phase-only objects by introducing a type of optical staining via an amplitude filter placed in the illumination path that consists of two or more colors. In this paper, the complete theory of Rheinberg illumination is derived, from which an algorithm is proposed that can digitally simulate the technique. Results are shown for a number of quantitative phase images of diatom cells obtained via digital holographic microscopy. The results clearly demonstrate the potential of the technique for label-free color staining of subcellular features.

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“…These observation methods require replacement of optical elements: a special objective lens for phase-contrast imaging and an iris diagraph. Rheinberg illumination microscopy enables simultaneous acquisition of bright-field and dark-field images using two-color illumination [14,15]. The central passing region of the condenser filter is used for bright field illumination.…”

Section: Introductionmentioning

confidence: 99%

Acquisition of Multi-Modal Images of Structural Modifications in Glass with Programmable LED-Array-Based Illumination

2019

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Ultrashort laser pulses can induce structural modifications in bulk glass, leading to refractive index change and scattering damage. As bright-field, dark-field, and phase imaging each provide complementary information about laser-induced structures, it is often desired to use multiple observations simultaneously. As described herein, we present the acquisition of bright-field, dark-field, and differential phase-contrast images of structural modifications induced in glass by femtosecond laser pulses with an LED array microscope. The contrast of refractive index change can be enhanced by differential phase-contrast images. We also report on the simultaneous acquisition of bright-field and dark-field images of structural modifications in a glass with LED-array-based Rheinberg illumination. A single-shot color image is separated to obtain bright field and dark field images simultaneously. We provide an experimental demonstration on multi-modal imaging of structural modifications in a glass with an LED array microscope using temporally-coded illumination and color-coded illumination.

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Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement

Cited by 24 publications

References 18 publications

Smart Computational Light Microscopes (SCLMs) of Smart Computational Imaging Laboratory (SCILab)

Smart Computational Light Microscopes (SCLMs) of Smart Computational Imaging Laboratory (SCILab)

Label-free color staining of quantitative phase images of biological cells by simulated Rheinberg illumination

Acquisition of Multi-Modal Images of Structural Modifications in Glass with Programmable LED-Array-Based Illumination

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