Digital inline holography of biological specimens

Ryle, James P.; Gopinathan, Unnikrishnan; McDonnell, Susan; Naughton, Thomas J.; Sheridan, John T.

doi:10.1117/12.680798

Cited by 10 publications

(12 citation statements)

References 10 publications

(6 reference statements)

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“…A DIHM system was designed, developed and purposely built to image MCF-7 and MDA-MB-231 mammalian cancer cell cultures 12,14 . This was originally done because of its compact footprint compared to the Mach-Zehnder or Michelson interferometers.…”

Section: Dihm Imaging Communication Optical Fibermentioning

confidence: 99%

Real world objects: capturing using in-line digital holography, projecting using spatial light modulators

et al. 2009

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Digital holography is the process where an object's phase and intensity information is retrieved from intensity images obtained using a digital camera (CCD or CMOS sensor). Unlike off-axis holography, object information is not modulated onto carrier fringes, thus in-line digital holography makes optimum use of the recording device's sampling bandwidth resulting in higher resolution digital holograms. However, reconstructed images are obscured by the linear superposition of the unwanted out of focus twin images. In addition to this, speckle noise degrades overall quality of the reconstructed images. The speckle effect is an optical phenomenon arising when laser sources are used in digital holographic systems. Minimizing the effects due to speckle noise, removal of the twin image and using the full sampling bandwidth of the capture device, aids overall reconstructed image quality. Using interferometric techniques, it is possible to record whole field information about an object. Digital processing of the reconstructed holograms can remove or suppress the twin image while effects from speckle noise can also be reduced numerically. Machine vision techniques can then be applied to segment and distinguish objects of interest in the hologram. Coding the resulting phase information onto a spatial light modulator (SLM), real world, three dimensional images of objects can be reconstructed using the computer generated hologram.

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Section: Dihm Imaging Communication Optical Fibermentioning

confidence: 99%

Real world objects: capturing using in-line digital holography, projecting using spatial light modulators

et al. 2009

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“…However, this lens-based magnification suffers from aberrations, which have to be corrected. Lensless digital holography is a well-established microscopic method providing diffraction limited resolution in the order of the wavelength of the used light source [4][5][6][7][8]. The Gabor inline configuration does not allow the separation of the different parts of the hologram after the reconstruction, so that DC part, real and twin images overlap.…”

Section: Introductionmentioning

confidence: 99%

Quantitative phase imaging by wide field lensless digital holographic microscope

et al. 2015

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Wide field, lensless microscopes have been developed for telemedicine and for resource limited setting [1]. They are based on in-line digital holography which is capable to provide amplitude and phase information resulting from numerical reconstruction. The phase information enables achieving axial resolution in the nanometer range. Hence, such microscopes provide a powerful tool to determine three-dimensional topologies of microstructures.In this contribution, a compact, low-cost, wide field, lensless microscope is presented, which is capable of providing topological profiles of microstructures in transparent material. Our setup consist only of two main components: a CMOSsensor chip and a laser diode without any need of a pinhole. We use this very simple setup to record holograms of microobjects. A wide field of view of ~24 mm², and a lateral resolution of ~2 µm are achieved. Moreover, amplitude and phase information are obtained from the numerical reconstruction of the holograms using a phase retrieval algorithm together with the angular spectrum propagation method. Topographic information of highly transparent micro-objects is obtained from the phase data. We evaluate our system by recording holograms of lines with different depths written by a focused laser beam. A reliable characterization of laser written microstructures is crucial for their functionality. Our results show that this system is valuable for determination of topological profiles of microstructures in transparent material.

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“…Since its serendipitous inception in the late 1940's, Gabor's holographic principle [1] has been used in the arena of electron holography [2], [3] the study of micro-organism trajectory [4], imaging microspheres [5], [6], cell cultures [7], as well as particle tracking in a volume [8]. Holographic imaging has be achieved using primary sources of various degrees of spatial coherence [ 6], [9][10][11][12].…”

Section: Introductionmentioning

confidence: 99%

Multispectral lensless digital in-line holographic microscope: LED illumination

2010

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Holography is the process where two coherent wavefields interfere resulting in an interference pattern from which whole field information can be retrieved. Digital holography is the process where the intensity of the superposition of the two waves is recorded using a light sensitive opto-electronic detector array such as a CCD or CMOS camera. From this recorded hologram it is possible to numerically reconstruct the object wavefield.When an optical beam is focused on a pinhole whose diameter is of the order of a few times the wavelength of the illumination beam, a spherically divergent wavefield is emitted. We use the emitted optical beam to illuminate weakly scattering objects resulting in a geometrically magnified diffraction pattern at the camera face. Scattered light from the object is the called the object wavefield, while unscattered light acts as the reference wavefield. The hologram is captured digitally before numerical reconstruction is applied to yield whole field information about the object.It is possible to reconstruct the objects wavefield using this method from coherent laser or incoherent LED illumination. The emitted light from the pinhole acts a pointsource of spatially coherent light enabling holography. This, in combination with the use of multiple wavelength LED's multispectral amplitude images can be reconstructed.The multispectral lensless DIHM described here can be used to holographically image biological specimens such as cells grown for use in the biopharmaceutical industry or for research purposes. In analysing cell viability based on the trypan blue assay, the outer membrane of non-viable cells is penetrated by violet blue dye. Using such a Digital In-line Holographic Microscope as described here, automatic classification of viable and non-viable cells could be performed.

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Digital inline holography of biological specimens

Cited by 10 publications

References 10 publications

Real world objects: capturing using in-line digital holography, projecting using spatial light modulators

Real world objects: capturing using in-line digital holography, projecting using spatial light modulators

Quantitative phase imaging by wide field lensless digital holographic microscope

Multispectral lensless digital in-line holographic microscope: LED illumination

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