Influence of defocus on quantitative analysis of microscopic objects and individual cells with digital holography.
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 of the measurement. Although holographic images can be refocused digitally using Fresnel propagation, ambiguity can result if this aspect is not carefully controlled. We demonstrate that microscopic objects can be accurately profiled by employing a digital refocusing method to analyze phase profiles of polystyrene microspheres and red blood cells.
Duke Scholars
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- 5102 Atomic, molecular and optical physics
- 4003 Biomedical engineering
- 3212 Ophthalmology and optometry
- 0912 Materials Engineering
- 0205 Optical Physics
Citation
Published In
DOI
EISSN
ISSN
Publication Date
Volume
Issue
Start / End Page
Related Subject Headings
- 5102 Atomic, molecular and optical physics
- 4003 Biomedical engineering
- 3212 Ophthalmology and optometry
- 0912 Materials Engineering
- 0205 Optical Physics