Three-dimensional Information and Refractive Index Measurement Using a Dual-wavelength Digital Holographic

In this work, for the first time, the quality of restoration in wide-field microscopy images after deconvolution was analyzed as a function of different Point Spread Functions using one deconvolution method, on a specimen of known size and on a biological specimen. The empirical Point Spread Function determination can significantly depend on the numerical aperture, refractive index of the embedding medium, refractive index of the immersion oil and cover slip thickness. The influence of all of these factors is shown in the same article and using the same microscope. We have found that the best deconvolution results are obtained when the empirical PSF utilized is obtained under the same conditions as the specimen. We also demonstrated that it is very important to quantitatively check the process' outcome using several quality indicators: Full-Width at Half-Maximum, Contrast-to-Noise Ratio, Signal-to-Noise Ratio and a Tenengrad-based function. We detected a significant improvement when using an indicator to measure the focus of the whole stack. Therefore, to qualitatively determinate the best deconvolved image between different conditions, one approach that we are pursuing is to use Tenengrad-based function indicators in images obtained using a wide-field microscope.

Section: Discussionmentioning

confidence: 88%

A Quantitative Study of the Quality of Deconvolved Wide-field Microscopy Images as Function of Empirical Three-dimensional Point Spread Functions

Adur

Vicente

Zamboni

et al. 2011

“…Digital holographic microscopy, because of the direct access to the phase profile, provides highly efficient and versatile phase imaging modality and it is undergoing rapid development. Subnanometer resolution of optical thickness has been demonstrated [13], the refractive index and physical thickness can be separated from the optical thickness [66], and optical phase unwrapping using three LEDs of different wavelengths has been demonstrated [67,68]. The ability to emulate various microscopy methodsincluding bright field, dark field, phase contrast, and DIC, from DHM has also been demonstrated [69].…”

Section: Quantitative Phase Microscopy By Digital Holography (Dhmentioning

confidence: 98%

Applications of Digital Holography in Biomedical Microscopy

Kim

2010

Digital holography (DH) is a potentially disruptive new technology for many areas of imaging science, especially in microscopy and metrology. DH offers a number of significant advantages such as the ability to acquire holograms rapidly, availability of complete amplitude and phase information of the optical field, and versatility of the interferometric and image processing techniques. This article provides a review of the digital holography, with an emphasis on its applications in biomedical microscopy. The quantitative phase microscopy by DH is described including some of the special techniques such as optical phase unwrapping and holography of total internal reflection. Tomographic imaging by digital interference holography (DIH) and related methods is described, as well as its applications in ophthalmic imaging and in biometry. Holographic manipulation and monitoring of cells and cellular components is another exciting new area of research. We discuss some of the current issues, trends, and potentials.

“…This method of digital recording and reconstruction of a numerical hologram is known as digital holography [7][8][9][10]. Digital holography has many advantages; for example, it does not require any chemical processing because the reconstructed image can be observed easily on a computer monitor, and numerical data can be obtained for three-dimensional (3D) objects [11,12].…”

Section: Introductionmentioning

confidence: 99%

Dual-wavelength Digital Holography Microscope for BGA Measurement Using Partial Coherence Sources

Cho¹,

Kim²,

Yu³

et al. 2011

Self Cite

Dual-wavelength holography has a better axial range than single-wavelength holography, allowing unambiguous phase imaging. Partial coherence sources reduce coherent noise, resulting in improved reconstructed images. We measured a ball-grid array using dual-wavelength holography with partial coherence sources. This holography method is useful for measurement samples that exhibit coherence noise and have a step height larger than the single wavelength used in holography.