A new flow cytometry method enabling rapid purification of diatoms from silica-rich lacustrine sediments

Tennant, Richard K.; Jones, Richard T.; Love, John; Lee, Robert J.

doi:10.1007/s10933-012-9659-5

Cited by 5 publications

(13 citation statements)

References 17 publications

(18 reference statements)

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“…The dating results from Lake Biwa were evaluated against those derived from terrestrial plant leaves originating from the same depths (table S2) as the pollen concentrates. The results from Lake Mondsee were evaluated against the core chronology (34) derived from varve counting and wiggle matching of the Lake Mondsee ostracod-derived  18 O record to the GICC05 chronology of the NGRIP  18 O record (35).…”

Section: Sediment Samplesmentioning

confidence: 99%

“…Potential applications comprise preanalytical concentration and purification of microfossils (e.g., pollen, diatoms, and algae) from sediment sequences that are valuable for the reconstruction of past environmental and climate changes and human-environment interactions. Different studies have successfully tested or positively evaluated the potential of conventional particle sorters for purifying diatoms (18) and pollen (19) for stable isotope analysis and microalgae (20) and pollen (21) for ancient DNA analysis. However, the size of these microfossils varies substantially and often exceeds the particle size limit (100 m) of available high-speed sorting devices.…”

Section: Introductionmentioning

confidence: 99%

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Breakthrough in purification of fossil pollen for dating of sediments by a new large-particle on-chip sorter

et al. 2021

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Particle sorting is a fundamental method in various fields of medical and biological research. However, existing sorting applications are not capable for high-throughput sorting of large-size (>100 micrometers) particles. Here, we present a novel on-chip sorting method using traveling vortices generated by on-demand microjet flows, which locally exceed laminar flow condition, allowing for high-throughput sorting (5 kilohertz) with a record-wide sorting area of 520 micrometers. Using an activation system based on fluorescence detection, the method successfully sorted 160-micrometer microbeads and purified fossil pollen (maximum dimension around 170 micrometers) from lake sediments. Radiocarbon dates of sorting-derived fossil pollen concentrates proved accurate, demonstrating the method’s ability to enhance building chronologies for paleoenvironmental records from sedimentary archives. The method is capable to cover urgent needs for high-throughput large-particle sorting in genomics, metabolomics, and regenerative medicine and opens up new opportunities for the use of pollen and other microfossils in geochronology, paleoecology, and paleoclimatology.

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Section: Sediment Samplesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Breakthrough in purification of fossil pollen for dating of sediments by a new large-particle on-chip sorter

et al. 2021

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“…Clay minerals can be removed by using the “heavy-liquid separation method”, by utilizing the difference of specific gravity of clay minerals and diatom frustules . In addition, diatoms can be separated from contaminants using gravitational split-flow thin fractionation, which separates particles of different densities in a laminar flow. − Recently, a new method for rapid extraction of diatom frustules from lacustrine sediment using flow cytometry was reported . The principles of flow cytometry are explained in more detail later in the Materials and Methods section.…”

Section: Introductionmentioning

confidence: 99%

“…The cell sorter can sort microparticles on the basis of optical characteristics of the subject, such as the size, internal structure, fluorescence intensity, or wavelength, at high speed (tens of thousands of particles per second in the instrument that we use), and is used for the analysis or selective sorting of cells. Recently, this technique has been applied to the separation of microbes, micromanganese nodules, and diatom frustules from sediment samples. ,, The aim of this study was to obtain accurate, taxon-specific δ 18 O records that can be applied to the various oceanographic settings in the Southern Ocean. We therefore targeted Thalassiosira lentiginosa (T.…”

Section: Introductionmentioning

confidence: 99%

Purification of Disc-Shaped Diatoms from the Southern Ocean Sediment by a Cell Sorter to Obtain an Accurate Oxygen Isotope Record

Ijiri

Izumi

Morono

et al. 2021

ACS Earth Space Chem.

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The oxygen isotopic composition (δ18O) of diatom frustules is potentially a quantitative paleoenvironmental proxy. However, δ18O analysis has some uncertainties because of difficulties in removing clay-mineral contamination and taxon-specific separation due to the small size of diatom frustules. The clay-mineral contamination seriously affects δ18O measurements, and the δ18O of bulk diatoms contains a mixture of information from diatoms living in various seasons and water depths. To resolve such problems, we established a new method for purifying the frustules of disc-shaped diatoms using a cell sorter and succeeded in removing the clay-mineral contamination and restricting the number of species for the δ18O analysis. We developed this method using samples from a sediment core retrieved from the Conrad Rise in the Indian Ocean sector of the Southern Ocean. First, the disc-shaped diatoms were enriched and cleaned by sieving, elutriation, and heavy-liquid separation. The samples were then further purified by a cell sorter. Regardless of the diatom assemblage or the sediment type in the core sample, this method generated samples consisting of more than 95% disc-shaped diatoms, dominated by Thalassiosira lentiginosa (Janisch) Fryxell, which occurs ubiquitously throughout the Southern Ocean. The δ18O profile for the sediment core obtained from purified disc-shaped diatoms showed the last glacial cycle since ca. 40 ka, which can be correlated with the LR04 benthic δ18O stack record and the temperature record in the EPICA Dome C ice core, indicating that this method can contribute to obtaining an accurate δ18O record of diatoms associated with glacial–interglacial cycles.

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“…We therefore used flow cytometry (FC) to rapidly distinguish, sort and collect the putative Botryococcus microfossils from defined sediment horizons, as previously performed for pollen grains 28 and diatom frustules 29 . Preserved DNA was extracted from this material and sequenced using next generation Illumina sequencing.…”

Section: Introductionmentioning

confidence: 99%

Palaeogenomics of the Hydrocarbon Producing Microalga Botryococcus braunii

Tennant

Lux

Sambles

et al. 2019

Sci Rep

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Botryococcus braunii is a colonial microalga that appears early in the fossil record and is a sensitive proxy of environmental and hydroclimatic conditions. Palaeozoic Botryococcus fossils which contribute up to 90% of oil shales and approximately 1% of crude oil, co-localise with diagnostic geolipids from the degradation of source-signature hydrocarbons. However more recent Holocene sediments demonstrate no such association. Consequently, Botryococcus are identified in younger sediments by morphology alone, where potential misclassifications could lead to inaccurate paleoenvironmental reconstructions. Here we show that a combination of flow cytometry and ancient DNA (aDNA) sequencing can unambiguously identify Botryococcus microfossils in Holocene sediments with hitherto unparalleled accuracy and rapidity. The application of aDNA sequencing to microfossils offers a far-reaching opportunity for understanding environmental change in the recent geological record. When allied with other high-resolution palaeoenvironmental information such as aDNA sequencing of humans and megafauna, aDNA from microfossils may allow a deeper and more precise understanding of past environments, ecologies and migrations.

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A new flow cytometry method enabling rapid purification of diatoms from silica-rich lacustrine sediments

Abstract: The version presented here may differ from the published version. If citing, you are advised to consult the published version for pagination, volume/issue and date of publication A new flow cytometry method enabling rapid purification of diatoms from silicarich lacustrine sediments.

Cited by 5 publications

References 17 publications

Breakthrough in purification of fossil pollen for dating of sediments by a new large-particle on-chip sorter

Breakthrough in purification of fossil pollen for dating of sediments by a new large-particle on-chip sorter

Purification of Disc-Shaped Diatoms from the Southern Ocean Sediment by a Cell Sorter to Obtain an Accurate Oxygen Isotope Record

Palaeogenomics of the Hydrocarbon Producing Microalga Botryococcus braunii

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