2015
DOI: 10.1364/boe.6.002067
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Influence of defocus on quantitative analysis of microscopic objects and individual cells with digital holography

Abstract: Digital holography offers a unique method for studying microscopic objects using quantitative measurements of the optical phase delays of transmitted light. The optical phase may be integrated across the object to produce an optical volume measurement, a parameter related to dry mass by a simple scaling factor. While digital holography is useful for comparing the properties of microscopic objects, especially cells, we show here that quantitative comparisons of optical phase can be influenced by the focal plane… Show more

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Cited by 43 publications
(42 citation statements)
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“…A diffraction phase microscope with a fluorescent arm capable of acquiring multiple fluorescent bands, as well as FRET signals, was constructed . The new QPI instrument employed a spectrally selected illumination of 670 nm with a bandwidth of 1.2 nm using a coherent white light source . Instead of using a typical off‐axis style interferometer arrangement, a diffraction phase interferometer was implemented.…”
Section: Methodsmentioning
confidence: 99%
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“…A diffraction phase microscope with a fluorescent arm capable of acquiring multiple fluorescent bands, as well as FRET signals, was constructed . The new QPI instrument employed a spectrally selected illumination of 670 nm with a bandwidth of 1.2 nm using a coherent white light source . Instead of using a typical off‐axis style interferometer arrangement, a diffraction phase interferometer was implemented.…”
Section: Methodsmentioning
confidence: 99%
“…During this study, OV was calculated to show dynamic changes in cell morphology during cell death, as cell volume regulation has been shown to be critically implicated in the activation of apoptosis . OV was calculated as follows: OV()t=λ2πx,ynormalΔϕ()titalicdxdy …”
Section: Methodsmentioning
confidence: 99%
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“…Moreover, DHM offers significant advantages over conventional microscopy in accessibility to quantitative amplitude and phase information, and in flexible digital processing such as storage, filtering, auto-focusing, aberration compensation, and display. DHM has been widely used in many fields such as biomedical science [2][3][4][5][6][7], micro-and nano-fabrication [8][9][10], materials science [11][12][13], the particle field [14][15][16][17], atomic physics [18], and the thermal energy field [19]. Nevertheless, the resolution and image quality of the most commonly used pre-magnification digital holographic microscopy (Pre-MDHM) and post-magnification digital holographic microscopy (Post-MDHM) are restricted by the photosensitive dimensions and the pixel size of the sensor, which has greatly limited the application of DHM.…”
Section: Introductionmentioning
confidence: 99%