Studying Glycolytic Oscillations in Individual Yeast Cells by Combining Fluorescence Microscopy with Microfluidics and Optical Tweezers

Gustavsson, Anna-Karin; Banaeiyan, Amin A.; Niekerk, David D. van; Snoep, Jacky L.; Adiels, Caroline B.; Goksör, Mattias

doi:10.1002/cpcb.70

Cited by 3 publications

(3 citation statements)

References 68 publications

(90 reference statements)

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“…Yeast cells, a kind of single-cell eukaryotic, are used as a typical specimen for optical micromanipulation. [32,33] To demonstrate that the proposed method is effective and independent of the cell type, yeast cells (average diameter: 5.5 µm), as representative immotile target cells, were first used in the experiment as a controllable cellular micromotor. Colloidal silica (SiO 2 ) particles, which are widely used in biomedicine for their good biocompatibility and many other intriguing characteristics, were chosen as the orbiting particles (diameter: 7.53 µm).…”

Section: Controllable Rotation Of Yeast Cellsmentioning

confidence: 99%

Controllable Cellular Micromotors Based on Optical Tweezers

Zou

Zheng

et al. 2020

Adv Funct Materials

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Micromotors hold exciting prospects in biomedical applications but still face a great challenge. To date, there have been few reports of micromotors with high safety, flexible controllability, and full biocompatibility. Here, a multifunctional method based on an optical tweezer system is presented to realize controllable cellular micromotors. The method not only satisfies all of the above criteria but is also independent of the cell types and materials. Optical tweezers are used to generate a dynamic scanning optical trap along a given circular trajectory, which can trap and drive a microparticle or a single cell to move along the trajectory and thus generate a microvortex. Cells within the microvortex will be controllably rotated under an action of shear stress or torque and their rotation rate and direction can be controlled by changing the scanning frequency and direction of the dynamic optical trap. The proposed method is effective for both immotile target cells and swimming target cells. Additionally, it is further applied to realize synchronous translation and rotation of cellular micromotors and to assemble controllable and fully biocompatible cellular micromotor assays. The proposed method is believed to have potential applications in targeted drug delivery, biological microenvironment monitoring and sensing, and biomedical treatment.

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Section: Controllable Rotation Of Yeast Cellsmentioning

confidence: 99%

Controllable Cellular Micromotors Based on Optical Tweezers

Zou

Zheng

et al. 2020

Adv Funct Materials

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“…Prior to experiments, PDMS chips with microfluidic channels were prepared using SU8 molds 27,28,29,30,31 . The SU8 molds were prepared by spin-coating approximately 5 mL of the photoresist (Kayaku Advanced Materials, Inc., Cat.…”

Section: Pdms (Polydimethylsiloxane) Microfluidic Chip Preparationmentioning

confidence: 99%

Lysine Demethylase 4A is a Centrosome Associated Protein Required for Centrosome Integrity and Genomic Stability

Chowdhury,

Wang,

Love

et al. 2024

Preprint

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Centrosomes play a fundamental role in nucleating and organizing microtubules in the cell and are vital for faithful chromosome segregation and maintenance of genomic stability. Loss of structural or functional integrity of centrosomes causes genomic instability and is a driver of oncogenesis. The lysine demethylase 4A (KDM4A) is an epigenetic ‘eraser’ of chromatin methyl marks, which we show also localizes to the centrosome with single molecule resolution. We additionally discovered KDM4A demethylase enzymatic activity is required to maintain centrosome homeostasis, and is required for centrosome integrity, a new functionality unlinked to altered expression of genes regulating centrosome number. We find rather, that KDM4A interacts with both mother and daughter centriolar proteins to localize to the centrosome in all stages of mitosis. Loss ofKDM4Aresults in supernumerary centrosomes and accrual of chromosome segregation errors including chromatin bridges and micronuclei, markers of genomic instability. In summary, these data highlight a novel role for an epigenetic ‘eraser’ regulating centrosome integrity, mitotic fidelity, and genomic stability at the centrosome.

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“…Our microfluidic chips consist of a polydimethylsiloxane (PDMS) channel [36][37][38][39][40][41] with a 3D nanoprinted metalized insert which reflects the LS into the sample and enables convenient perfusion of solutions during multi-target Exchange-PAINT imaging. Previously developed single-objective LS systems have utilized various reflection mechanisms such as a micromirror 42 , an AFM cantilever mirror 43 , pyramidal microcavities 44 , a glass prism 45 , and a microfluidic chamber with metalized side walls 46 to redirect the LS into the sample.…”

Section: Introductionmentioning

confidence: 99%

Whole-cell multi-target single-molecule super-resolution imaging in 3D with microfluidics and a single-objective tilted light sheet

Saliba,

Gagliano,

Gustavsson

2023

Preprint

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Multi-target single-molecule super-resolution fluorescence microscopy offers a powerful means of understanding the distributions and interplay between multiple subcellular structures at the nanoscale. However, single-molecule super-resolution imaging of whole mammalian cells is often hampered by high fluorescence background and slow acquisition speeds, especially when imaging multiple targets in 3D. In this work, we have mitigated these issues by developing a steerable, dithered, single-objective tilted light sheet for optical sectioning to reduce fluorescence background and a pipeline for 3D nanoprinting microfluidic systems for reflection of the light sheet into the sample and for efficient and automated solution exchange. By combining these innovations with PSF engineering for nanoscale localization of individual molecules in 3D, deep learning for analysis of overlapping emitters, active 3D stabilization for drift correction and long-term imaging, and Exchange-PAINT for sequential multi-target imaging without chromatic offsets, we demonstrate whole-cell multi-target 3D single-molecule super-resolution imaging with improved precision and imaging speed.

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Studying Glycolytic Oscillations in Individual Yeast Cells by Combining Fluorescence Microscopy with Microfluidics and Optical Tweezers

Cited by 3 publications

References 68 publications

Controllable Cellular Micromotors Based on Optical Tweezers

Controllable Cellular Micromotors Based on Optical Tweezers

Lysine Demethylase 4A is a Centrosome Associated Protein Required for Centrosome Integrity and Genomic Stability

Whole-cell multi-target single-molecule super-resolution imaging in 3D with microfluidics and a single-objective tilted light sheet

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