Brillouin flow cytometry for label-free mechanical phenotyping of the nucleus

Zhang, Jitao; Nou, Xuefei A.; Kim, Hanyoup; Scarcelli, Giuliano

doi:10.1039/c6lc01443g

Cited by 71 publications

(68 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The overall elastic stiffness of retinal tissue measured in this study was comparable to that of other CNS tissues assessed by AFM (6, 8–11, 47, 50–52). Similarly, the Brillouin shifts we measured in the retina are comparable with values published in previous literature (18, 20, 21). …”

Section: Discussionsupporting

confidence: 92%

The role of cell body density in ruminant retina mechanics assessed by atomic force and Brillouin microscopy

et al. 2017

Self Cite

View full text Add to dashboard Cite

Cells in the central nervous system (CNS) respond to the stiffness of their environment. CNS tissue is mechanically highly heterogeneous, thus providing motile cells with region-specific mechanical signals. While CNS mechanics has been measured with a variety of techniques, reported values of tissue stiffness vary greatly, and the morphological structures underlying spatial changes in tissue stiffness remain poorly understood. We here exploited two complementary techniques, contact-based atomic force microscopy and contact-free Brillouin microscopy, to determine the mechanical properties of ruminant retinae, which are built up by different tissue layers. As in all vertebrate retinae, layers of high cell body densities (‘nuclear layers’) alternate with layers of low cell body densities (‘plexiform layers’). Different tissue layers varied significantly in their mechanical properties, with the photoreceptor layer being the stiffest region of the retina, and the inner plexiform layer belonging to the softest regions. As both techniques yielded similar results, our measurements allowed us to calibrate the Brillouin microscopy measurements and convert the Brillouin shift into a quantitative assessment of elastic tissue stiffness with optical resolution. Similar as in the mouse spinal cord and the developing Xenopus brain, we found a strong correlation between nuclear densities and tissue stiffness. Hence, the cellular composition of retinae appears to strongly contribute to local tissue stiffness, and Brillouin microscopy shows a great potential for the application in vivo to measure the mechanical properties of transparent tissues.

show abstract

Section: Discussionsupporting

confidence: 92%

The role of cell body density in ruminant retina mechanics assessed by atomic force and Brillouin microscopy

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Imaging Approaches For Dynamic Platformsmentioning

confidence: 99%

“…Extraction of the low signals of the Brillouin shifts in comparison to the strong elastic signal is a technical challenge for arranging this imaging technique. Mechanical properties of the cell population at the throughput of 200 cells per hour can be extracted in a subcellular range (Figure j) …”

Section: Imaging Approaches For Dynamic Platformsmentioning

confidence: 99%

“…Mechanical properties of the cell population at the throughput of 200 cells per hour can be extracted in a subcellular range (Figure 25j). [121] For a better intuition, Figure 25 depicts examples of output images of the aforementioned techniques. Moreover, Table 3 and Figure 26 represent the main features and a comparison of the scattering-based techniques for dynamic bioplatforms, respectively.…”

Section: Wwwsmall-journalcommentioning

confidence: 99%

See 1 more Smart Citation

Optical Imaging Approaches to Monitor Static and Dynamic Cell‐on‐Chip Platforms: A Tutorial Review

Arandian

Bagheri

Ehtesabi

et al. 2019

Small

View full text Add to dashboard Cite

Miniaturized laboratories on chip platforms play an important role in handling life sciences studies. The platforms may contain static or dynamic biological cells. Examples are a fixed medium of an organ‐on‐a‐chip and individual cells moving in a microfluidic channel, respectively. Due to feasibility of control or investigation and ethical implications of live targets, both static and dynamic cell‐on‐chip platforms promise various applications in biology. To extract necessary information from the experiments, the demand for direct monitoring is rapidly increasing. Among different microscopy methods, optical imaging is a straightforward choice. Considering light interaction with biological agents, imaging signals may be generated as a result of scattering or emission effects from a sample. Thus, optical imaging techniques could be categorized into scattering‐based and emission‐based techniques. In this review, various optical imaging approaches used in monitoring static and dynamic platforms are introduced along with their optical systems, advantages, challenges, and applications. This review may help biologists to find a suitable imaging technique for different cell‐on‐chip studies and might also be useful for the people who are going to develop optical imaging systems in life sciences studies.

show abstract

“…In contrast, microfluidic devices enable highthroughput measurements of nuclear and cellular mechanics with precisely defined geometries. [28][29][30] Some microfluidic devices measure the stiffness of cells based on their transit time when perfused through narrow constrictions [31][32][33][34] or mimic micropipette aspiration, 35 but these approaches are often hampered by clogging due to particles, large cell aggregates, or cell adhesion in the constrictions. This problem can be alleviated in devices that use fluid shear stress to deform the cells rather than constrictions, 36 but the deformations achieved in these devices do not recapitulate the extensive deformations that can be achieved using physical barriers.…”

Section: Introductionmentioning

confidence: 99%

High-throughput microfluidic micropipette aspiration device to probe time-scale dependent nuclear mechanics in intact cells

Davidson

Fedorchak

Mondésert-Deveraux

et al. 2019

Preprint

View full text Add to dashboard Cite

The mechanical properties of the cell nucleus are increasingly recognized as critical in many biological processes. The deformability of the nucleus determines the ability of immune and cancer cells to migrate through tissues and across endothelial cell layers, and changes to the mechanical properties of the nucleus can serve as novel biomarkers in processes such as cancer progression and stem cell differentiation. However, current techniques to measure the viscoelastic nuclear mechanical properties are often time consuming, limited to probing one cell at a time, or require expensive, highly specialized equipment. Furthermore, many current assays do not measure time-dependent properties, which are characteristic of viscoelastic materials. Here, we present an easy-to-use microfluidic device that applies the well-established approach of micropipette aspiration, adapted to measure many cells in parallel. The device design allows rapid loading and purging of cells for measurements, and minimizes clogging by large particles or clusters of cells. Combined with a semiautomated image analysis pipeline, the microfluidic device approach enables significantly increased experimental throughput. We validated the experimental platform by comparing computational models of the fluid mechanics in the device with experimental measurements of fluid flow. In addition, we conducted experiments on cells lacking the nuclear envelope protein lamin A/C and wild-type controls, which have well-characterized nuclear mechanical properties. Fitting timedependent nuclear deformation data to power law and different viscoelastic models revealed that loss of lamin A/C significantly altered the elastic and viscous properties of the nucleus, resulting in substantially increased nuclear deformability. Lastly, to demonstrate the versatility of the devices, we characterized the viscoelastic nuclear mechanical properties in a variety of cell lines and experimental model systems, including human skin fibroblasts from an individual with a mutation in the lamin gene associated with dilated cardiomyopathy, healthy control fibroblasts, induced pluripotent stem cells (iPSCs), and human tumor cells. Taken together, these experiments demonstrate the ability of the microfluidic device and automated image analysis platform to provide robust, high throughput measurements of nuclear mechanical properties, including time-dependent elastic and viscous behavior, in a broad range of applications.

show abstract

Brillouin flow cytometry for label-free mechanical phenotyping of the nucleus

Cited by 71 publications

References 52 publications

The role of cell body density in ruminant retina mechanics assessed by atomic force and Brillouin microscopy

The role of cell body density in ruminant retina mechanics assessed by atomic force and Brillouin microscopy

Optical Imaging Approaches to Monitor Static and Dynamic Cell‐on‐Chip Platforms: A Tutorial Review

High-throughput microfluidic micropipette aspiration device to probe time-scale dependent nuclear mechanics in intact cells

Contact Info

Product

Resources

About