3-Step flow focusing enables multidirectional imaging of bioparticles for imaging flow cytometry

Kleiber, Andreas; Ramoji, Anuradha; Mayer, Günter; Neugebauer, Ute; Henkel, Thomas

doi:10.1039/d0lc00244e

Cited by 18 publications

(22 citation statements)

References 40 publications

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“…Though allowing rapid assessments across thousands of pollen grains, impedance FCM was also reported to overestimate pollen viability rates when compared to direct in vitro pollen germination tests [55], suggesting that the main strength of this approach could be in extensive pollen fertility screening across numerous individuals (e.g., in agriculture or breeding practice). ise to expand this application [58,59]. In addition, FCM data could prove to be a valuable complement to genetic methods for the identification of species, for example, pollen barcoding [60], that provide information about species composition but not abundance.…”

Section: Counts and Proportions Of Particle Typesmentioning

confidence: 99%

“…Studies of this kind are limited by the extent to which the measured trait, such as DNA content or autofluorescence, can distinguish between potential candidate taxa. However, recent developments in instrumentation, including multi‐directional imaging, airflow cytometry, and impedance cytometry, as well as analytical techniques such as 3D image analysis and neural networks, promise to expand this application [58, 59]. In addition, FCM data could prove to be a valuable complement to genetic methods for the identification of species, for example, pollen barcoding [60], that provide information about species composition but not abundance.…”

Section: Research Applications In Pollen Fcmmentioning

confidence: 99%

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Flow cytometric analysis of pollen and spores: An overview of applications and methodology

et al. 2021

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Pollen grains are the male gametophytes in a seed-plant life cycle. Their small, particulate nature and crucial role in plant reproduction have made them an attractive object of study using flow cytometry (FCM), with a wide range of applications existing in the literature. While methodological considerations for many of these overlap with those for other tissue types (e.g., general considerations for the measurement of nuclear DNA content), the relative complexity of pollen compared to single cells presents some unique challenges. We consider these here in the context of both the identification and isolation of pollen and its subunits, and the types of research applications. While the discussion here mostly concerns pollen, the general principles described here can be extended to apply to spores in ferns, lycophytes, and bryophytes. In addition to recommendations provided in more general studies, some recurring and notable issues related specifically to pollen and spores are highlighted.

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Section: Counts and Proportions Of Particle Typesmentioning

confidence: 99%

Section: Research Applications In Pollen Fcmmentioning

confidence: 99%

Flow cytometric analysis of pollen and spores: An overview of applications and methodology

et al. 2021

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“…Recently, Kleiber et al (2020) reported the three-step hydrodynamic flow focusing of cells for applications in imaging flow cytometry, which operates at low Re <10 and self-alignment of all cells within the focal plane of the imaging system [38,39]. Combined with a subsequent microfluidic sorter unit the resulting system match the target use case.…”

Section: Introductionmentioning

confidence: 99%

Three step flow focusing enables image-based discrimination and sorting of late stage 1 Haematococcus pluvialis cells

et al. 2021

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Label-free and gentle separation of cell stages with desired target properties from mixed stage populations are a major research task in modern biotechnological cultivation process and optimization of micro algae. The reported microfluidic sorter system (MSS) allows the subsequent investigation of separated subpopulations. The implementation of a viability preserving MSS is shown for separation of late stage 1 Haematococcus pluvialis (HP) cells form a mixed stage population. The MSS combines a three-step flow focusing unit for aligning the cells in single file transportation mode at the center of the microfluidic channel with a pure hydrodynamic sorter structure for cell sorting. Lateral displacement of the cells into one of the two outlet channels is generated by piezo-actuated pump chambers. In-line decision making for sorting is based on a user-definable set of image features and properties. The reported MSS significantly increased the purity of target cells in the sorted population (94%) in comparison to the initial mixed stage population (19%).

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“…Many emerging technologies in the recent years aim at surpassing the abovementioned limitations. The most promising approaches regard the label‐free imaging methods combined with artificial intelligence (AI) algorithms 13‐16 . Indeed, several scientific papers based on the sole label‐free techniques, such as quantitative phase imaging (QPI), were able to distinguish between tumor cell and correspondent nontransformed cell line, thus restricting the results to a special class.…”

Section: Introductionmentioning

confidence: 99%

“…At this scope, AI is hungry for informative data and, in the context of cell type discrimination, high‐throughput QPI is the most appropriate candidate to satisfy this need. Indeed, many valuable papers describe the possibility to classify different cell types by combining QPI with AI in the field of tumor cell identification 16‐20 . In this framework, one of the main developing fields for the CTCs detection and isolation regards microfluidic engineering and its ingenious solutions able to perform both in marker‐based and label‐free systems 21‐27 …”

Section: Introductionmentioning

confidence: 99%

Perspectives on liquid biopsy for label‐free detection of “circulating tumor cells” through intelligent lab‐on‐chips

Miccio

Cimmino

Kurelac

et al. 2020

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Circulating tumor cells (CTCs) are rare tumor cells released from primary, metastatic, or recurrent tumors in the peripheral blood of cancer patients. CTCs isolation from peripheral blood and their molecular characterization represent a new marker in cancer screening, a diagnostic tool called “liquid biopsy” (LB). Compared to traditional tissue biopsy that is invasive and does not reveal tumor heterogeneity, LB is noninvasive and reflects in “real‐time” tumor dynamism and drug sensitivity. In the frame of LB, a new paradigm based on single‐cell and label‐free analysis based on morphological analysis is emerging. Here, we review the latest research developments in this emerging vision of LB. In particular, we survey and discuss recent improvements in microfluidics, imaging label‐free diagnosis and cell classification by artificial intelligence and how to combine them to realize an intelligent platform based on lab‐on‐chip technology. This prospect appears to open up promising and intriguing new scenarios for cancer management through single‐cell analysis that will revolutionize the future of early cancer diagnosis and therapeutic choice with disruptive impact on the society.

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3-Step flow focusing enables multidirectional imaging of bioparticles for imaging flow cytometry

Abstract: The control of the focus plane allows multi-directional imaging flow cytometry.

Cited by 18 publications

References 40 publications

Flow cytometric analysis of pollen and spores: An overview of applications and methodology

Flow cytometric analysis of pollen and spores: An overview of applications and methodology

Three step flow focusing enables image-based discrimination and sorting of late stage 1 Haematococcus pluvialis cells

Perspectives on liquid biopsy for label‐free detection of “circulating tumor cells” through intelligent lab‐on‐chips

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