Digital Holography and Cell Studies

Alm, Kersti; Cirenajwis, Helena; Gisselsson, Lennart; Wingren, Anette Gjörloff; Janicke, Birgit; Mölder, Anna; Oredsson, Stina; Persson, Johan

doi:10.5772/15364

Cited by 23 publications

(18 citation statements)

References 64 publications

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“…There have been some significant advances recently in non-invasive, label-free imaging1. Examples of these include digital holographic microscopy234567 and ptychography8910.…”

mentioning

confidence: 99%

Ptychography – a label free, high-contrast imaging technique for live cells using quantitative phase information

Marrison

Räty

Marriott

et al. 2013

Sci Rep

146

116

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Cell imaging often relies on synthetic or genetic fluorescent labels, to provide contrast which can be far from ideal for imaging cells in their in vivo state. We report on the biological application of a, label-free, high contrast microscopy technique known as ptychography, in which the image producing step is transferred from the microscope lens to a high-speed phase retrieval algorithm. We demonstrate that this technology is appropriate for label-free imaging of adherent cells and is particularly suitable for reporting cellular changes such as mitosis, apoptosis and cell differentiation. The high contrast, artefact-free, focus-free information rich images allow dividing cells to be distinguished from non-dividing cells by a greater than two-fold increase in cell contrast, and we demonstrate this technique is suitable for downstream automated cell segmentation and analysis.

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“…There have been some significant advances recently in non-invasive, label-free imaging1. Examples of these include digital holographic microscopy234567 and ptychography8910.…”

mentioning

confidence: 99%

Ptychography – a label free, high-contrast imaging technique for live cells using quantitative phase information

Marrison

Räty

Marriott

et al. 2013

Sci Rep

146

116

View full text Add to dashboard Cite

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“…In addition, DHM has particularly been proven as a powerful technique in microbiology, where it can be used to assess parameters such as thickness, volume and refractive index of living as well as dead cells [8,9]. This non--invasive and staining--less technique has therefore helped elucidate cell movement in both 2D and 3D, as well as shed light on cell morphology, both at a single cell level and for entire populations [10].…”

Section: Introductionmentioning

confidence: 99%

Cell shape identification using digital holographic microscopy

2015

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We present a cost--effective, simple and fast digital holographic microscopy method based upon Rayleigh--Sommerfeld back propagation for identification of the geometrical shape of a cell. The method was tested using synthetic hologram images generated by ray--tracing software and from experimental images of semi--transparent spherical beads and living red blood cells. Our results show that by only using the real part of the back--reconstructed amplitude the proposed method can provide information of the geometrical shape of the object and at the same time accurately determine the axial position of the object under study. The proposed method can be used in flow chamber assays for pathophysiological studies where fast morphological changes of cells are studied in high numbers and at different heights.

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“…The importance of RI measurements for biology is well underlined in a work that it is one of the first introducing Quantitative Phase Microscopy (QPM) [14], presenting results for the thickness and RI of airway smooth muscle (ASM) cells. Due to the difficulties of QPM to decouple evaluation of thickness and RI, both involved in the evaluated optical path difference (OPD), a further procedure is presented based on digital holographic microscopy (DHM) [15] founded on a dual wavelength observation, as for other successive works [16,17]. Here, the KK equation is part of the needed formalism.…”

Section: Introductionmentioning

confidence: 99%

Phase and Index of Refraction Imaging by Hyperspectral Reflectance Confocal Microscopy

Selci

2016

Molecules

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A hyperspectral reflectance confocal microscope (HSCM) was realized by CNR-ISC (Consiglio Nazionale delle Ricerche-Istituto dei Sistemi Complessi) a few years ago. The instrument and data have been already presented and discussed. The main activity of this HSCM has been within biology, and reflectance data have shown good matching between spectral signatures and the nature or evolution on many types of cells. Such a relationship has been demonstrated mainly with statistical tools like Principal Component Analysis (PCA), or similar concepts, which represent a very common approach for hyperspectral imaging. However, the point is that reflectance data contains much more useful information and, moreover, there is an obvious interest to go from reflectance, bound to the single experiment, to reflectivity, or other physical quantities, related to the sample alone. To accomplish this aim, we can follow well-established analyses and methods used in reflectance spectroscopy. Therefore, we show methods of calculations for index of refraction n, extinction coefficient k and local thicknesses of frequency starting from phase images by fast Kramers-Kronig (KK) algorithms and the Abeles matrix formalism. Details, limitations and problems of the presented calculations as well as alternative procedures are given for an example of HSCM images of red blood cells (RBC).

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Digital Holography and Cell Studies

Cited by 23 publications

References 64 publications

Ptychography – a label free, high-contrast imaging technique for live cells using quantitative phase information

Ptychography – a label free, high-contrast imaging technique for live cells using quantitative phase information

Cell shape identification using digital holographic microscopy

Phase and Index of Refraction Imaging by Hyperspectral Reflectance Confocal Microscopy

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