Molecular phenotyping of aging in single yeast cells using a novel microfluidic device

Xie, Zhengwei; Zhang, Yi; Zou, Ke; Brandman, Onn; Luo, Chunxiong; Ouyang, Qi; Li, Hao

doi:10.1111/j.1474-9726.2012.00821.x

Cited by 110 publications

(138 citation statements)

References 39 publications

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“…Recently, such devices have been designed that enable the tracking of yeast cells throughout their lifespan, making it possible to record and study cellular phenotypic changes during aging (21)(22)(23). However, many issues prevent the use of microfluidic devices in a high-throughput manner for lifespan screens.…”

Section: Saccharomyces Cerevisiaementioning

confidence: 99%

“…Fourth, the micropad design often allows more than one cell to be trapped; multiple cells can be trapped underneath one micropad, whereas no cells are trapped under others. Finally, in one of the designs, cell-surface labeling and chemical modification of the device are required, which has proven to be technically challenging for fabrication and to introduce adverse effects on replicative lifespan (23).…”

Section: Saccharomyces Cerevisiaementioning

confidence: 99%

“…However, many issues prevent the use of microfluidic devices in a high-throughput manner for lifespan screens. First, although the time required to monitor the entire lifespan of the yeast cell has been dramatically reduced, the throughput is limited to 1-4 channels per device (21)(22)(23). Second, mother cells were immobilized underneath soft elastomer [polydimethylsiloxane (PDMS)] micropads (21,22).…”

Section: Saccharomyces Cerevisiaementioning

confidence: 99%

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High-throughput analysis of yeast replicative aging using a microfluidic system

Liu

et al. 2015

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

150

174

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Saccharomyces cerevisiae has been an important model for studying the molecular mechanisms of aging in eukaryotic cells. However, the laborious and low-throughput methods of current yeast replicative lifespan assays limit their usefulness as a broad genetic screening platform for research on aging. We address this limitation by developing an efficient, high-throughput microfluidic single-cell analysis chip in combination with high-resolution time-lapse microscopy. This innovative design enables, to our knowledge for the first time, the determination of the yeast replicative lifespan in a highthroughput manner. Morphological and phenotypical changes during aging can also be monitored automatically with a much higher throughput than previous microfluidic designs. We demonstrate highly efficient trapping and retention of mother cells, determination of the replicative lifespan, and tracking of yeast cells throughout their entire lifespan. Using the high-resolution and large-scale data generated from the high-throughput yeast aging analysis (HYAA) chips, we investigated particular longevity-related changes in cell morphology and characteristics, including critical cell size, terminal morphology, and protein subcellular localization. In addition, because of the significantly improved retention rate of yeast mother cell, the HYAA-Chip was capable of demonstrating replicative lifespan extension by calorie restriction.microfluidics | high-throughput | replicative aging | calorie restriction | Saccharomyces cerevisiae

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Section: Saccharomyces Cerevisiaementioning

confidence: 99%

Section: Saccharomyces Cerevisiaementioning

confidence: 99%

Section: Saccharomyces Cerevisiaementioning

confidence: 99%

See 1 more Smart Citation

High-throughput analysis of yeast replicative aging using a microfluidic system

Liu

et al. 2015

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

150

174

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“…To connect the asymmetric division in young cells to the aging phenotype in old cells, we tracked how these proteins change over time in single mother cells throughout their lifespan, using a recently developed microfluidic device (32,33). We analyzed Did2p and Tpo3p, which represent a protein involved in endosomal sorting/endosome-to-vacuole transport and a nitrogen source transporter, respectively.…”

Section: Mother-enriched and Lifespan-limiting Proteins Tend To Accummentioning

confidence: 99%

Systematic analysis of asymmetric partitioning of yeast proteome between mother and daughter cells reveals “aging factors” and mechanism of lifespan asymmetry

Yang

McCormick

Zheng

et al. 2015

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

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Budding yeast divides asymmetrically, giving rise to a mother cell that progressively ages and a daughter cell with full lifespan. It is generally assumed that mother cells retain damaged, lifespan limiting materials ("aging factors") through asymmetric division. However, the identity of these aging factors and the mechanisms through which they limit lifespan remain poorly understood. Using a flow cytometry-based, high-throughput approach, we quantified the asymmetric partitioning of the yeast proteome between mother and daughter cells during cell division, discovering 74 mother-enriched and 60 daughterenriched proteins. While daughter-enriched proteins are biased toward those needed for bud construction and genome maintenance, mother-enriched proteins are biased towards those localized in the plasma membrane and vacuole. Deletion of 23 of the 74 motherenriched proteins leads to lifespan extension, a fraction that is about six times that of the genes picked randomly from the genome. Among these lifespan-extending genes, three are involved in endosomal sorting/endosome to vacuole transport, and three are nitrogen source transporters. Tracking the dynamic expression of specific mother-enriched proteins revealed that their concentration steadily increases in the mother cells as they age, but is kept relatively low in the daughter cells via asymmetric distribution. Our results suggest that some mother-enriched proteins may increase to a concentration that becomes deleterious and lifespan-limiting in aged cells, possibly by upsetting homeostasis or leading to aberrant signaling. Our study provides a comprehensive resource for analyzing asymmetric cell division and aging in yeast, which should also be valuable for understanding similar phenomena in other organisms.aging | asymmetric cell division | proteome C ellular aging and asymmetric cell division are intimately linked. In budding yeast, asymmetric cell division yields a mother cell and a daughter cell that are easily distinguishable under the microscope. Tracking the fate of the mother lineage led to the discovery that individual mother cells have a finite replicative lifespan, defined by the number of daughters a mother cell produced before senescence (1). It is known that although the mother cell ages with each division, their daughters retain the same full lifespan independent of the age of the mother at least until the last few mother cell divisions (2, 3). Thus, the asymmetry in cell division leads to asymmetry of aging.Even in single-celled organisms in which cell division is seemingly morphologically symmetric, such as fission yeast or Escherichia coli, asymmetric partitioning of cellular contents can still occur and have a differential impact on the aging/death fate of the two offspring (4-8). Asymmetric cell division is also a general phenomenon in mammalian cells (e.g., during development or in mitotically active tissues), where cell division typically leads to two cells with distinct fates, often with different replicative potential. It has been argued...

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“…[18] They attached the yeast cells on a wall coated by avidin to track individual mother cells throughout their whole lifespan, and found that the aging of cells was characterized by an increased general stress and a progressive lengthening of cell cycle for the last few cell divisions. [19] The Walkmoto group demonstrated that growth noise among single cells could cause clonal populations of Escherichia coli to divide faster than the mean doubling time of their constituent single cells. [20] Recently, with the help of a "mother machine" device, the Jacobs-Wagner group discovered that microorganisms like E. coli and C. crescentus achieved cell size homeostasis by growing the same volume or length between divisions.…”

Section: High Resolutionmentioning

confidence: 99%

Applications of Microfluidics in Quantitative Biology

Bai

Gao

Wen

et al. 2017

Biotechnology Journal

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Quantitative biology is dedicated to taking advantage of quantitative reasoning and advanced engineering technologies to make biology more predictable. Microfluidics, as an emerging technique, provides new approaches to precisely control fluidic conditions on small scales and collect data in highthroughput and quantitative manners. In this review, the authors present the relevant applications of microfluidics to quantitative biology based on two major categories (channel-based microfluidics and droplet-based microfluidics), and their typical features. We also envision some other microfluidic techniques that may not be employed in quantitative biology right now, but have great potential in the near future.

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Molecular phenotyping of aging in single yeast cells using a novel microfluidic device

Cited by 110 publications

References 39 publications

High-throughput analysis of yeast replicative aging using a microfluidic system

High-throughput analysis of yeast replicative aging using a microfluidic system

Systematic analysis of asymmetric partitioning of yeast proteome between mother and daughter cells reveals “aging factors” and mechanism of lifespan asymmetry

Applications of Microfluidics in Quantitative Biology

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