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Partial dark-field microscopy for investigating domain structures of double-layer microsphere film.

ABSTRACT: A lateral dislocation in a double-layer microsphere film is very difficult to identify because the constituent domains have the same two-dimensional crystalline orientation. Orientation-sensitive optical techniques cannot resolve this issue. Here, we demonstrate that partial dark-field (pDF) optical microscopy can be very effective in identifying this type of domain boundary and dislocation of a close-packed microsphere double-layer. Using the hexagonal symmetry of the close-packed microsphere film and the light-focusing property of microspheres, the partially blocked dark-field condenser can provide much higher contrast than other optical microscopy modes can in identifying the laterally dislocated domains. The former can also distinguish domains with different crystalline orientation by rotating the pDF stop. The simplicity of the pDF mode will make it an ideal tool for the structural study of close-packed double-layer microsphere films.

SUBMITTER: Heon Kim J

PROVIDER: S-EPMC4426674 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature

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Partial dark-field microscopy for investigating domain structures of double-layer microsphere film.

Heon Kim Joon J Su Park Jung J

Scientific reports 20150511

A lateral dislocation in a double-layer microsphere film is very difficult to identify because the constituent domains have the same two-dimensional crystalline orientation. Orientation-sensitive optical techniques cannot resolve this issue. Here, we demonstrate that partial dark-field (pDF) optical microscopy can be very effective in identifying this type of domain boundary and dislocation of a close-packed microsphere double-layer. Using the hexagonal symmetry of the close-packed microsphere f ...[more]

PMID: 25959375

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Project description:BackgroundThe haemozoin crystal continues to be investigated extensively for its potential as a biomarker for malaria diagnostics. In order for haemozoin to be a valuable biomarker, it must be present in detectable quantities in the peripheral blood and distinguishable from false positives. Here, dark-field microscopy coupled with sophisticated image processing algorithms is used to characterize the abundance of detectable haemozoin within infected erythrocytes from field samples in order to determine the window of detection in peripheral blood.MethodsThin smears from Plasmodium falciparum-infected and uninfected patients were imaged in both dark field (DF) unstained and bright field (BF) Giemsa-stained modes. The images were co-registered such that each parasite had thumbnails in both BF and DF modes, providing an accurate map between parasites and DF objects. This map was used to find the abundance of haemozoin as a function of parasite stage through careful parasite staging and correlation with DF objects. An automated image-processing and classification algorithm classified the bright spots in the DF images as either haemozoin or non-haemozoin objects.ResultsThe algorithm distinguishes haemozoin from non-haemozoin objects in DF images with an object-level sensitivity of 95% and specificity of 97%. Ring stages older than about 6 hours begin to show detectable haemozoin, and rings between 10-16 hours reliably contain detectable haemozoin. However, DF microscopy coupled with the image-processing algorithm detect no haemozoin in rings younger than six hours.DiscussionAlthough this method demonstrates the most sensitive detection of haemozoin in field samples reported to date, it does not detect haemozoin in ring-stage parasites younger than six hours. Thus, haemozoin is a poor biomarker for field samples primarily composed of young ring-stage parasites because the crystal is not present in detectable quantities by the methods described here. Based on these results, the implications for patient-level diagnosis and recommendations for future work are discussed.

Dataset Information

Partial dark-field microscopy for investigating domain structures of double-layer microsphere film.

Publications

Partial dark-field microscopy for investigating domain structures of double-layer microsphere film.

Similar Datasets

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