Fabrication of high-aspect-ratio amorphous perfluorinated polymer structure for total internal reflection fluorescence microscopy

Nakahara, Asahi; Shirasaki, Yoshitaka; Kawai, Kentaro; Ohara, Osamu; Mizuno, Jun; Shoji, Shuichi

doi:10.1016/j.mee.2011.02.117

Cited by 8 publications

(9 citation statements)

References 6 publications

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“…Signal / background noise ratio of the simple PDMS microwell array was 1.40 and that of the PDMS-CYTOP hybrid microwell was 4.51. The PDMS-CYTOP hybrid microwell showed equivalent TIRF performance of microwell fabricated with CYTOP reported in our previous work [5].…”

Section: Tirf Microscopysupporting

confidence: 64%

“…We have already reported about the fabrication method of microwell array device made of an amorphous perfluoropolymer: CYTOP which have identical refractive index [4], and it was shown that CYTOP microstructures did not interfere with TIRF imaging [5,6]. However, CYTOP structures fabricated by the method were non-uniform in height due to the shrinkage in volume of CYTOP during cure process (Figure 1 (c)).…”

Section: Introductionmentioning

confidence: 99%

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PDMS-CYTOP hybrid structure microwell array chip for total internal reflection fluorescence microscopy

Nakahara

Shirasaki

Mizuno

et al. 2013

The 8th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems

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A microwell arrry device with a hybrid structure of amorphour perfluoroplymer, CYTOP TM , and polydimethylsiloxane (PDMS) which is specialized in total internal refllection fluorescence (TIRF) microscopy was developed. Since the CYTOP layer, whose refractive index is as same as that of water, is sandwiched between PDMS and glass substrate, leakage of incident light is avoided. PDMS layer upon the CYTOP enables high aspect ratio structures and easy integration of microfluidic components. Since direct bonding of CYTOP and PDMS is difficult to achieve, a chemical bonding method using aminosilane coupling reagent and 3-Glycoidoxypropyltriethoxysilane was applied. The bond strength was about 0.74 MPa. 24 × 50 microwell array was fabricated and their optical properties was evaluated.Measured TIRF microscopy results shows no interfere with incident light under the conventionally used conditions.

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Section: Tirf Microscopysupporting

confidence: 64%

Section: Introductionmentioning

confidence: 99%

PDMS-CYTOP hybrid structure microwell array chip for total internal reflection fluorescence microscopy

Nakahara

Shirasaki

Mizuno

et al. 2013

The 8th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems

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“…With fluorescence-based, dynamic imaging of biological samples, the resolution and the image quality of membrane processes rely heavily on refractive index matching between the sample and the surrounding medium. Materials such as CYTOP 58 and MyPolymer 59 have similar refractive indices to water, which would allow super-resolution imaging, and this would give us…”

Section: Surface Modification Of Polymers For Membrane Fusionmentioning

confidence: 99%

Membrane-coated 3D architectures for bottom-up synthetic biology

et al. 2021

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“…In addition, we considered the following two points in developing the real‐time monitoring system: (i) intracellular events can also be monitored, when necessary, in parallel with the detection of secreted molecules and (ii) cells can be retrieved at a specified timing. For these purposes, a new microwell device equipped with total internal reflection illumination was produced (13). While methodological details of this method will be reported later (Shirasaki et al, manuscript in preparation), this new system, for the first time, enables simultaneous monitoring of cytokine secretion and intracellular events (anything seen on the same microscopy platform, such as translocation of transcription factors, membrane intactness, changes in Ca 2+ concentration, etc.)…”

Section: Real‐time Monitoring Of Cytokine Secretion From Single Immunmentioning

confidence: 99%

Toward an understanding of immune cell sociology: real‐time monitoring of cytokine secretion at the single‐cell level

et al. 2012

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The immune system is a very complex and dynamic cellular system, and its intricacies are considered akin to those of human society. Disturbance of homeostasis of the immune system results in various types of diseases; therefore, the homeostatic mechanism of the immune system has long been a subject of great interest in biology, and a lot of information has been accumulated at the cellular and the molecular levels. However, the sociological aspects of the immune system remain too abstract to address because of its high complexity, which mainly originates from a large number and variety of cell-cell interactions. As long-range interactions mediated by cytokines play a key role in the homeostasis of the immune system, cytokine secretion analyses, ranging from analyses of the micro level of individual cells to the macro level of a bulk of cell ensembles, provide us with a solid basis of a sociological viewpoint of the immune system. In this review, as the first step toward a comprehensive understanding of immune cell sociology, cytokine secretion of immune cells is surveyed with a special emphasis on the single-cell level, which has been overlooked but should serve as a basis of immune cell sociology. Now that it has become evident that large cell-tocell variations in cytokine secretion exist at the single-cell level, we face a tricky yet interesting question: How is homeostasis maintained when the system is composed of intrinsically noisy agents? In this context, we discuss how the heterogeneity of cytokine secretion at the single-cell level affects our view of immune cell sociology. While the apparent inconsistency between homeostasis and cell-to-cell heterogeneity is difficult to address by a conventional reductive approach, comparison and integration of single-cell data with macroscopic data will offer us a new direction for the comprehensive understanding of immune cell sociology. V C 2012 IUBMB Life, 65(1): [28][29][30][31][32][33][34] 2013

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Fabrication of high-aspect-ratio amorphous perfluorinated polymer structure for total internal reflection fluorescence microscopy

Cited by 8 publications

References 6 publications

PDMS-CYTOP hybrid structure microwell array chip for total internal reflection fluorescence microscopy

PDMS-CYTOP hybrid structure microwell array chip for total internal reflection fluorescence microscopy

Membrane-coated 3D architectures for bottom-up synthetic biology

Toward an understanding of immune cell sociology: real‐time monitoring of cytokine secretion at the single‐cell level

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