Using the Gouy phase anomaly to localize and track bacteria in digital holographic microscopy 4D images

Gibson, T. F.; Bedrossian, Manuel; Serabyn, Eugene; Lindensmith, C. A.; Nadeau, Jay L.

doi:10.1364/josaa.404004

Cited by 8 publications

(4 citation statements)

References 31 publications

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“…We use DHM in an 'inline' configuration, which can be implemented easily on a commercial microscope [11]. DHM has been used to track swimming microorganisms including bacteria [12][13][14][15][16], archaea [17], dinoflagellates [18], sperm cells [19][20][21], and other eukaryotes [22,23]. In the inline configuration, a sample chamber is illuminated with coherent light, part of which is scattered by the objects of interest.…”

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confidence: 99%

The chiral beat of algal flagella: force and torque via imaging

Wilson¹,

Bees²

2022

Preprint

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Flagella allow eukaryotic cells to move and pump fluid. We present the first three-dimensional, time-resolved imaging of the flagellar waveform of Chlamydomonas reinhardtii, a model alga found in fresh water. During the power stroke, we find that the flagella show rotational symmetry about the cell's centreline, but during the recovery stroke they display mirror symmetry about the same axis. We use our three-dimensional imaging data to test the applicability of resistive force theory when a force-free configuration is approximated using a cell on a micropipette. The inferred values of cells' swimming speeds and rotation rates show good agreement with experimental results on freely swimming cells.

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confidence: 99%

The chiral beat of algal flagella: force and torque via imaging

Wilson¹,

Bees²

2022

Preprint

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“…These sequences of images were calculated using the Rayleigh-Sommerfeld back-propagation scheme ( Lee and Grier, 2007 ). We localised the cells using a method based on the Gouy phase anomaly, as described in more detail elsewhere ( Wilson and Zhang, 2012 ; Farthing et al, 2017 ; Gibson et al, 2021 ). This method segments features based on axial optical intensity gradients within a sample, allowing us to extract three-dimensional coordinates for individual cells in each frame.…”

Section: Methodsmentioning

confidence: 99%

High-speed, three-dimensional imaging reveals chemotactic behaviour specific to human-infective Leishmania parasites

Findlay

Osman

Spence

et al. 2021

eLife

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Cellular motility is an ancient eukaryotic trait, ubiquitous across phyla with roles in predator avoidance, resource access, and competition. Flagellar motility is seen in various parasitic protozoans, and morphological changes in flagella during the parasite life cycle have been observed. We studied the impact of these changes on motility across life cycle stages, and how such changes might serve to facilitate human infection. We used holographic microscopy to image swimming cells of different Leishmania mexicana life cycle stages in three dimensions. We find that the human-infective (metacyclic promastigote) forms display ‘run and tumble’ behaviour in the absence of stimulus, reminiscent of bacterial motion, and that they specifically modify swimming direction and speed to target host immune cells in response to a macrophage-derived stimulus. Non-infective (procyclic promastigote) cells swim more slowly, along meandering helical paths. These findings demonstrate adaptation of swimming phenotype and chemotaxis towards human cells.

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“…For the aluminum oxide beads and gas vesicles, holograms were reconstructed in phase, and z-localization was performed using a z-derivative approach as described previously [ 49 ].…”

Section: Methodsmentioning

confidence: 99%

Microscopic Object Classification through Passive Motion Observations with Holographic Microscopy

Rouzie

Lindensmith

Nadeau

2021

Life

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Digital holographic microscopy provides the ability to observe throughout a volume that is large compared to its resolution without the need to actively refocus to capture the entire volume. This enables simultaneous observations of large numbers of small objects within such a volume. We have constructed a microscope that can observe a volume of 0.4 µm × 0.4 µm × 1.0 µm with submicrometer resolution (in xy) and 2 µm resolution (in z) for observation of microorganisms and minerals in liquid environments on Earth and on potential planetary missions. Because environmental samples are likely to contain mixtures of inorganics and microorganisms of comparable sizes near the resolution limit of the instrument, discrimination between living and non-living objects may be difficult. The active motion of motile organisms can be used to readily distinguish them from non-motile objects (live or inorganic), but additional methods are required to distinguish non-motile organisms and inorganic objects that are of comparable size but different composition and structure. We demonstrate the use of passive motion to make this discrimination by evaluating diffusion and buoyancy characteristics of cells, styrene beads, alumina particles, and gas-filled vesicles of micron scale in the field of view.

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Using the Gouy phase anomaly to localize and track bacteria in digital holographic microscopy 4D images

Cited by 8 publications

References 31 publications

The chiral beat of algal flagella: force and torque via imaging

The chiral beat of algal flagella: force and torque via imaging

High-speed, three-dimensional imaging reveals chemotactic behaviour specific to human-infective Leishmania parasites

Microscopic Object Classification through Passive Motion Observations with Holographic Microscopy

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