Three-dimensional motion measurements of free-swimming microorganisms using digital holographic microscopy

Lee, Sang Joon; Seo, Kyung Won; Choi, Yong Seok; Sohn, Myung-Hwan

doi:10.1088/0957-0233/22/6/064004

Cited by 38 publications

(19 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…More advanced microscopy configurations (17)(18)(19) or holographic imaging schemes (20)(21)(22)(23)(24)(25)(26)(27)(28) have also been used to resolve 3D trajectories of sperms of other species. However, these previous approaches have not reported submicron 3D localization accuracy throughout a large observation volume of ≥1 μL.…”

Section: Discussionmentioning

confidence: 99%

“…Compared to earlier reports that also used holographic imaging techniques (20)(21)(22)(23)(24)(25)(26)(27)(28) to track sperms or other microorganisms, our approach is lensfree ( Fig. 1) and therefore exhibits a significantly larger imaging field-of-view of >17 mm 2 together with unit fringe magnification, while still achieving submicron positioning accuracy that is necessary to observe human sperms' tight helical paths.…”

mentioning

confidence: 95%

See 1 more Smart Citation

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

Ozcan

2012

Proc. Natl. Acad. Sci. U.S.A.

314

277

View full text Add to dashboard Cite

Dynamic tracking of human sperms across a large volume is a challenging task. To provide a high-throughput solution to this important need, here we describe a lensfree on-chip imaging technique that can track the three-dimensional (3D) trajectories of >1,500 individual human sperms within an observation volume of approximately 8-17 mm 3 . This computational imaging platform relies on holographic lensfree shadows of sperms that are simultaneously acquired at two different wavelengths, emanating from two partially-coherent sources that are placed at 45°with respect to each other. This multiangle and multicolor illumination scheme permits us to dynamically track the 3D motion of human sperms across a field-of-view of >17 mm 2 and depth-of-field of approximately 0.5-1 mm with submicron positioning accuracy. The large statistics provided by this lensfree imaging platform revealed that only approximately 4-5% of the motile human sperms swim along well-defined helices and that this percentage can be significantly suppressed under seminal plasma. Furthermore, among these observed helical human sperms, a significant majority (approximately 90%) preferred right-handed helices over left-handed ones, with a helix radius of approximately 0.5-3 μm, a helical rotation speed of approximately 3-20 rotations∕s and a linear speed of approximately 20-100 μm∕s. This high-throughput 3D imaging platform could in general be quite valuable for observing the statistical swimming patterns of various other microorganisms, leading to new insights in their 3D motion and the underlying biophysics.human sperm imaging | sperm tracking | digital holography | micro-swimmer O bserving three-dimensional (3D) trajectories of sperms is in general a challenging task. This is partially due to limited imaging volume of optical microscopes that are based on conventional lenses. For human sperms this becomes even more challenging since the sperm head is small (approximately 3-4 μm) demanding a relatively high-magnification objective lens, and moves rather fast (20-100 μm∕s) which makes it difficult to track their 3D swimming patterns as they quickly move out of the observation volume of an objective lens. Partly due to this low throughput and the limited spatial and temporal sampling windows that conventional microscopes provide, natural 3D swimming patterns of human sperms and their statistics could not be reported so far. Earlier results (1-19) that were obtained using lens-based conventional microscopes either measured the 2D trajectories of the human sperms along a focal plane, or reported on sperms of other species such as sea urchin, which were significantly easier to resolve under a microscope since their 3D rotation diameter is larger (>13 μm) together with a lower rotation frequency compared to human sperms.Here we report a new technique that is based on lensfree holographic imaging on a chip to dynamically track the 3D trajectories of human sperms across a large volume of approximately 8-17 mm 3 ( Fig. 1) with submicron positioning accuracy. Thi...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 95%

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

Ozcan

2012

Proc. Natl. Acad. Sci. U.S.A.

314

277

View full text Add to dashboard Cite

show abstract

“…Due to the large depth-of-focus, special numerical focus functions have to be used to exactly determine the depth position of the bio-sample. Lee et al (2011) reported the best results for a focus function that relies on the variation in the axial intensity gradient. They reported an rms uncertainty in the axial position of 21.5 lm for 30 lm particles.…”

Section: Digital Holographic Microscopymentioning

confidence: 99%

Particle imaging techniques for volumetric three-component (3D3C) velocity measurements in microfluidics

Cierpka

Kähler

2011

J Vis

131

View full text Add to dashboard Cite

The reliable measurement of the 3D3C velocity field in microfluidic devices becomes more and more important for future optimization and developments for lab-on-a-chip applications or point-of-care medical diagnosis systems. In the past years, different particle-based imaging methods, such as confocal scanning microscopy, holography, stereoscopic and tomographic imaging or approaches based on defocused particle images or optical aberrations have been developed and applied successfully to measure velocity fields in microfluidic systems. The benefits and drawbacks of these methods will be discussed in detail as the proper understanding of the measurement principle is essential to select the most appropriate technique for a desired measurement application. Once an imaging method is chosen, the velocity can be estimated by correlation-based methods or tracking approaches. The advantages and disadvantages of both methods and the importance of image preprocessing will also be discussed in detail.

show abstract

“…The challenge to generate such understanding is the three dimensional motion of spores at a velocity of &25 body lengths per second (150 lm/s) [14]. Holography, as first invented by Gabor in 1948 [15], is capable of imaging such fast, three-dimensional trajectories [16][17][18][19][20][21]. The physical concept of holography allows the capture of the complete information about objects within a given volume by encoding into an interference pattern.…”

Section: Introductionmentioning

confidence: 99%

Settlement Behavior of Zoospores of Ulva linza During Surface Selection Studied by Digital Holographic Microscopy

et al. 2012

View full text Add to dashboard Cite

Settlement of the planktonic dispersal stages of marine organisms is the crucial step for the development of marine biofouling. Four-dimensional holographic tracking reveals the mechanism by which algal spores select surfaces suitable for colonization. Quantitative analysis of the three dimensional swimming trajectories of motile spores of a macroalga (Ulva linza) in the vicinity of surfaces functionalized with different chemistries reveals that their search strategy and swimming behavior is correlated to the number of settled spores found in spore settlement bioassays conducted over 45 min. The spore motility and exploration behavior can be classified into different motion patterns, with their relative occurrence changing with the surface chemistry. Based on the detailed motility analysis we derived a model for the surface selection and settlement process of Ulva zoospores.

show abstract

Three-dimensional motion measurements of free-swimming microorganisms using digital holographic microscopy

Cited by 38 publications

References 20 publications

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

Particle imaging techniques for volumetric three-component (3D3C) velocity measurements in microfluidics

Settlement Behavior of Zoospores of Ulva linza During Surface Selection Studied by Digital Holographic Microscopy

Contact Info

Product

Resources

About