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Article ; Online: Optimizing Scanning Bessel Beam Light Sheet Microscopy with Custom-designed Lens Cap for Expansion Microscopy.

Lee, Chia-Ming / Tian, Xuejiao / Tsai, Min-Ju / Chen, Bi-Chang

Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada

2023 Volume 29, Issue Supplement_1, Page(s) 1004

Language	English
Publishing date	2023-08-23
Publishing country	England
Document type	Journal Article
ZDB-ID	1385710-1
ISSN	1435-8115 ; 1431-9276
ISSN (online)	1435-8115
ISSN	1431-9276
DOI	10.1093/micmic/ozad067.504
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Article: Macrophages Break Interneuromast Cell Quiescence by Intervening in the Inhibition of Schwann Cells in the Zebrafish Lateral Line.

Lin, Meng-Ju / Lee, Chia-Ming / Hsu, Wei-Lin / Chen, Bi-Chang / Lee, Shyh-Jye

Frontiers in cell and developmental biology

2022 Volume 10, Page(s) 907863

Abstract: In the zebrafish lateral line system, interneuromast cells (INCs) between neuromasts are kept quiescent by underlying Schwann cells (SWCs). Upon severe injuries that cause the complete loss of an entire neuromast, INCs can occasionally differentiate into ...

Abstract	In the zebrafish lateral line system, interneuromast cells (INCs) between neuromasts are kept quiescent by underlying Schwann cells (SWCs). Upon severe injuries that cause the complete loss of an entire neuromast, INCs can occasionally differentiate into neuromasts but how they escape from the inhibition by SWCs is still unclear. Using a genetic/chemical method to ablate a neuromast precisely, we found that a small portion of larvae can regenerate a new neuromast. However, the residual regeneration capacity was hindered by inhibiting macrophages. Using in toto imaging, we further discovered heterogeneities in macrophage behavior and distribution along the lateral line. We witnessed the crawling of macrophages between the injured lateral line and SWCs during regeneration and between the second primordium and the first mature lateral line during development. It implies that macrophages may physically alleviate the nerve inhibition to break the dormancy of INCs during regeneration and development in the zebrafish lateral line.
Language	English
Publishing date	2022-07-01
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2022.907863
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Article ; Online: Large-scale expanded sample imaging with tiling lattice lightsheet microscopy.

Lu, Chieh-Han / Huang, Cheng-Yu / Tian, Xuejiao / Chen, Peilin / Chen, Bi-Chang

The international journal of biochemistry & cell biology

2022 Volume 154, Page(s) 106340

Abstract: The ability to observe biological nanostructures forms a vital step in understanding their functions. Thanks to the invention of expansion microscopy (ExM) technology, super-resolution features of biological samples can now be easily visualized with ... ...

Abstract	The ability to observe biological nanostructures forms a vital step in understanding their functions. Thanks to the invention of expansion microscopy (ExM) technology, super-resolution features of biological samples can now be easily visualized with conventional light microscopies. However, when the sample is physically expanded, the demand for deep and precise 3D imaging increases. Lattice lightsheet microscopy (LLSM), which utilizes a planar illumination that is confined within the imaging depth of high numerical aperture (NA=1.1) detection objective, fulfils such requirements. In addition, optical tiling could be implemented to increase the field of view (FoV) by moving the lightsheet without mechanically moving the samples or the objective for high-precision 3D imaging. In this review article, we will explain the principle of the tiling lattice lightsheet microscopy (tLLSM), which combines optical tiling and lattice lightsheet, and discuss the applications of tLLSM in ExM.
MeSH term(s)	Microscopy/methods
Language	English
Publishing date	2022-11-26
Publishing country	Netherlands
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	1228429-4
ISSN	1878-5875 ; 1357-2725
ISSN (online)	1878-5875
ISSN	1357-2725
DOI	10.1016/j.biocel.2022.106340
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Zs.A 431: Show issues

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Article ; Online: Autism-related KLHL17 and SYNPO act in concert to control activity-dependent dendritic spine enlargement and the spine apparatus.

Hu, Hsiao-Tang / Lin, Yung-Jui / Wang, Ueh-Ting Tim / Lee, Sue-Ping / Liou, Yae-Huei / Chen, Bi-Chang / Hsueh, Yi-Ping

PLoS biology

2023 Volume 21, Issue 8, Page(s) e3002274

Abstract: Dendritic spines, the tiny and actin-rich protrusions emerging from dendrites, are the subcellular locations of excitatory synapses in the mammalian brain that control synaptic activity and plasticity. Dendritic spines contain a specialized form of ... ...

Abstract	Dendritic spines, the tiny and actin-rich protrusions emerging from dendrites, are the subcellular locations of excitatory synapses in the mammalian brain that control synaptic activity and plasticity. Dendritic spines contain a specialized form of endoplasmic reticulum (ER), i.e., the spine apparatus, required for local calcium signaling and that is involved in regulating dendritic spine enlargement and synaptic plasticity. Many autism-linked genes have been shown to play critical roles in synaptic formation and plasticity. Among them, KLHL17 is known to control dendritic spine enlargement during development. As a brain-specific disease-associated gene, KLHL17 is expected to play a critical role in the brain, but it has not yet been well characterized. In this study, we report that KLHL17 expression in mice is strongly regulated by neuronal activity and KLHL17 modulates the synaptic distribution of synaptopodin (SYNPO), a marker of the spine apparatus. Both KLHL17 and SYNPO are F-actin-binding proteins linked to autism. SYNPO is known to maintain the structure of the spine apparatus in mature spines and contributes to synaptic plasticity. Our super-resolution imaging using expansion microscopy demonstrates that SYNPO is indeed embedded into the ER network of dendritic spines and that KLHL17 is closely adjacent to the ER/SYNPO complex. Using mouse genetic models, we further show that Klhl17 haploinsufficiency and knockout result in fewer dendritic spines containing ER clusters and an alteration of calcium events at dendritic spines. Accordingly, activity-dependent dendritic spine enlargement and neuronal activation (reflected by extracellular signal-regulated kinase (ERK) phosphorylation and C-FOS expression) are impaired. In addition, we show that the effect of disrupting the KLHL17 and SYNPO association is similar to the results of Klhl17 haploinsufficiency and knockout, further strengthening the evidence that KLHL17 and SYNPO act together to regulate synaptic plasticity. In conclusion, our findings unravel a role for KLHL17 in controlling synaptic plasticity via its regulation of SYNPO and synaptic ER clustering and imply that impaired synaptic plasticity contributes to the etiology of KLHL17-related disorders.
MeSH term(s)	Animals ; Mice ; Actins ; Autistic Disorder/genetics ; Autistic Disorder/metabolism ; Brain ; Dendritic Spines ; Genes, fos ; Hypertrophy ; Microfilament Proteins/genetics ; Microfilament Proteins/metabolism
Chemical Substances	Actins ; Microfilament Proteins ; Synpo protein, mouse
Language	English
Publishing date	2023-08-31
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2126776-5
ISSN	1545-7885 ; 1544-9173
ISSN (online)	1545-7885
ISSN	1544-9173
DOI	10.1371/journal.pbio.3002274
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Article ; Online: Two-photon scanned light sheet fluorescence microscopy with axicon imaging for fast volumetric imaging.

Lin, Po-Yen / Hwang, Sheng-Ping L / Lee, Chi-Hon / Chen, Bi-Chang

Journal of biomedical optics

2021 Volume 26, Issue 11

Abstract: Significance: Two-photon microscopy has become the standard platform for deep-tissue fluorescence imaging. However, the use of point scanning in conventional two-photon microscopy limits the speed of volumetric image acquisition.: Aim: To obtain fast ...

Abstract	Significance: Two-photon microscopy has become the standard platform for deep-tissue fluorescence imaging. However, the use of point scanning in conventional two-photon microscopy limits the speed of volumetric image acquisition. Aim: To obtain fast and deep volumetric images, we combine two-photon light sheet fluorescence microscopy (2p-LSFM) and axicon imaging that yields an extended depth of field (DOF) in 2p-LSFM. Approach: Axicon imaging is achieved by imposing an axicon lens in the detection part of LSFM. Results: The DOF with axicon imaging is extended more than 20-fold over that of a conventional imaging lens, liberating the synchronized scanning in LSFM. We captured images of dynamic beating hearts and red blood cells in zebrafish larvae at volume acquisition rates up to 30 Hz. Conclusions: We demonstrate the fast three-dimensional imaging capability of 2p-LSFM with axicon imaging by recording the rapid dynamics of physiological processes.
MeSH term(s)	Animals ; Imaging, Three-Dimensional ; Lenses ; Microscopy, Fluorescence ; Zebrafish
Language	English
Publishing date	2021-11-15
Publishing country	United States
Document type	Journal Article
ZDB-ID	1309154-2
ISSN	1560-2281 ; 1083-3668
ISSN (online)	1560-2281
ISSN	1083-3668
DOI	10.1117/1.JBO.26.11.116503
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Zs.A 4699: Show issues

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Article ; Online: Recent advances in the use of fluorescent nanoparticles for bioimaging.

Pratiwi, Feby Wijaya / Kuo, Chiung Wen / Chen, Bi-Chang / Chen, Peilin

Nanomedicine (London, England)

2019 Volume 14, Issue 13, Page(s) 1759–1769

Abstract: Rapid and recent progress in fluorescence microscopic techniques has allowed for routine discovery and viewing of biological structures and processes in unprecedented spatiotemporal resolution. In these imaging techniques, fluorescent nanoparticles (NPs) ...

Abstract	Rapid and recent progress in fluorescence microscopic techniques has allowed for routine discovery and viewing of biological structures and processes in unprecedented spatiotemporal resolution. In these imaging techniques, fluorescent nanoparticles (NPs) play important roles in the improvement of reporting systems. A short overview of recently developed fluorescent NPs used for advanced
MeSH term(s)	Animals ; Fluorescent Dyes/analysis ; Humans ; Microscopy, Fluorescence/methods ; Nanoparticles/analysis ; Optical Imaging/methods ; Theranostic Nanomedicine/methods
Chemical Substances	Fluorescent Dyes
Language	English
Publishing date	2019-07-12
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2277839-1
ISSN	1748-6963 ; 1743-5889
ISSN (online)	1748-6963
ISSN	1743-5889
DOI	10.2217/nnm-2019-0105
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Zs.A 6617: Show issues

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Article ; Online: Protein and lipid expansion microscopy with trypsin and tyramide signal amplification for 3D imaging.

Wang, Ueh-Ting Tim / Tian, Xuejiao / Liou, Yae-Huei / Lee, Sue-Ping / Hu, Hsiao-Tang / Lu, Chieh-Han / Lin, Po-Ting / Cheng, Ya-Jen / Chen, Peilin / Chen, Bi-Chang

Scientific reports

2023 Volume 13, Issue 1, Page(s) 21922

Abstract: Expansion microscopy, whereby the relative positions of biomolecules are physically increased via hydrogel expansion, can be used to reveal ultrafine structures of cells under a conventional microscope. Despite its utility for achieving super-resolution ... ...

Abstract	Expansion microscopy, whereby the relative positions of biomolecules are physically increased via hydrogel expansion, can be used to reveal ultrafine structures of cells under a conventional microscope. Despite its utility for achieving super-resolution imaging, expansion microscopy suffers a major drawback, namely reduced fluorescence signals caused by excessive proteolysis and swelling effects. This caveat results in a lower photon budget and disfavors fluorescence imaging over a large field of view that can cover an entire expanded cell, especially in 3D. In addition, the complex procedures and specialized reagents of expansion microscopy hinder its popularization. Here, we modify expansion microscopy by deploying trypsin digestion to reduce protein loss and tyramide signal amplification to enhance fluorescence signal for point-scanning-based imaging. We name our new methodology TT-ExM to indicate dual trypsin and tyramide treatments. TT-ExM may be applied for both antibody and lipid staining. TT-ExM displayed enhanced protein retention for endoplasmic reticulum and mitochondrial markers in COS-7 cell cultures. Importantly, TT-ExM-based lipid staining clearly revealed the complex 3D membrane structures in entire expanded cells. Through combined lipid and DNA staining, our TT-ExM methodology highlighted mitochondria by revealing their DNA and membrane structures in cytoplasm, as well as the lipid-rich structures formed via phase separation in nuclei at interphase. We also observed lipid-rich chromosome matrices in the mitotic cells. These high-quality 3D images demonstrate the practicality of TT-ExM. Thus, readily available reagents can be deployed in TT-ExM to significantly enhance fluorescence signals and generate high-quality and ultrafine-resolution images under confocal microscopy.
MeSH term(s)	Trypsin ; Imaging, Three-Dimensional/methods ; Proteins ; Microscopy, Confocal/methods ; Indicators and Reagents ; DNA ; Lipids
Chemical Substances	Trypsin (EC 3.4.21.4) ; Proteins ; Indicators and Reagents ; DNA (9007-49-2) ; Lipids
Language	English
Publishing date	2023-12-08
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-48959-9
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Article ; Online: SARS-CoV-2 N protein mediates intercellular nucleic acid dispersion, a feature reduced in Omicron.

Wu, Jung-Lin / Kuan, I-I / Guo, Jing-You / Hsu, Wei-Chia / Tang, Wei-Chun / Chan, Hsin-Ju / Chen, Yu-Ju / Chen, Bi-Chang / Wu, Han-Chung / Liao, James C

iScience

2023 Volume 26, Issue 2, Page(s) 105995

Abstract: The coronavirus nucleocapsid (N) protein is known to bind to nucleic acids and facilitate viral genome encapsulation. Here we report that the N protein can mediate RNA or DNA entering neighboring cells through ACE2-independent, receptor (STEAP2)-mediated ...

Abstract	The coronavirus nucleocapsid (N) protein is known to bind to nucleic acids and facilitate viral genome encapsulation. Here we report that the N protein can mediate RNA or DNA entering neighboring cells through ACE2-independent, receptor (STEAP2)-mediated endocytosis, and achieve gene expression. The effect is more pronounced for the N protein of wild-type SARS-CoV-2 than that of the Omicron variant and other human coronaviruses. This effect is enhanced by RANTES (CCL5), a chemokine induced by N protein, and lactate, a metabolite produced in hypoxia, to cause more damage. These findings might explain the clinical observations in SARS-CoV-2-infected cases. Moreover, the N protein-mediated function can be inhibited by N protein-specific monoclonal antibodies or p38 mitogen-activated protein kinase inhibitors. Since the N-protein-mediated nucleic acid endocytosis involves a receptor commonly expressed in many types of cells, our findings suggest that N protein may have an additional role in SARS-CoV-2 pathogenesis.
Language	English
Publishing date	2023-01-16
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.105995
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Article: Simultaneous Single-Particle Tracking and Dynamic pH Sensing Reveal Lysosome-Targetable Mesoporous Silica Nanoparticle Pathways

Zhang, Rong-Lin / Pratiwi, Feby Wijaya / Chen, Bi-Chang / Chen, Peilin / Wu, Si-Han / Mou, Chung-Yuan

ACS applied materials & interfaces. 2020 July 13, v. 12, no. 38

2020

Abstract: Nanoparticle (NP)-based targeted drug delivery is intended to transport therapeutically active molecules to specific cells and particular intracellular compartments. However, there is limited knowledge regarding the complete route of NPs in this ... ...

Abstract	Nanoparticle (NP)-based targeted drug delivery is intended to transport therapeutically active molecules to specific cells and particular intracellular compartments. However, there is limited knowledge regarding the complete route of NPs in this targeting scenario. In this study, simultaneously performing motion and dynamic pH sensing using single-particle tracking (SPT) leads to an alternative method of gaining insights into the mesoporous silica nanoparticle’s (MSN) journey in targeting lysosome. Two different pH-sensitive dyes and a reference dye are incorporated into mesoporous silica nanoparticles (MSNs) via co-condensation to broaden the measurable pH range (pH 4–7.5) of the nanoprobe. The phosphonate, amine, and lysosomal sorting peptides (YQRLGC) are conjugated onto the MSN’s surface to study intracellular nano-biointeractions of two oppositely charged and lysosome-targetable MSNs. The brightness and stability of these MSNs allow their movement and dynamic pH evolution during their journey to be simultaneously monitored in real time. Importantly, a multidimensional analysis of MSN’s movement and local pH has revealed new model intracellular dynamic states and distributions of MSNs, previously inaccessible when using single parameters alone. A key result is that YQRLGC-conjugated MSNs took an alternative route to target lysosomes apart from the traditional one, which sped up to 4 h and enhanced their targeting efficiency (up to 32%). The findings enrich our understanding of the intracellular journey of MSNs. This study offers complementary information on correlating the surface design with the full pathway of nanoparticles to achieve targeted delivery of therapeutic payload.
Keywords	drugs ; dyes ; evolution ; lysosomes ; pH ; peptides ; phosphonates ; porous media ; silica ; therapeutics
Language	English
Dates of publication	2020-0713
Size	p. 42472-42484.
Publishing place	American Chemical Society
Document type	Article
Note	NAL-AP-2-clean
ISSN	1944-8252
DOI	10.1021/acsami.0c07917
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Lattice light sheet microscopy using tiling lattice light sheets.

Gao, Liang / Tang, Wei-Chun / Tsai, Yun-Chi / Chen, Bi-Chang

Optics express

2018 Volume 27, Issue 2, Page(s) 1497–1506

Abstract: We present a novel method used to implement tiling lattice light sheets (LLS) in lattice light sheet microscopy (LLSM) on regular LLS microscopes without changing the LLS microscope hardware. A LLS is tiled by applying binary phase maps acquired from off- ...

Abstract	We present a novel method used to implement tiling lattice light sheets (LLS) in lattice light sheet microscopy (LLSM) on regular LLS microscopes without changing the LLS microscope hardware. A LLS is tiled by applying binary phase maps acquired from off-center cross-sections of the corresponding optical lattice to the binary SLM used in LLS microscopes, by which a thin LLS can be tiled to image large specimens while maintaining the 3D imaging ability in the entire field of view. We investigate the method via numerical simulations and experiments, and demonstrate the method by imaging fluorescent particles embedded in agarose gel and expanded cells in the dithered mode of LLSM.
Language	English
Publishing date	2018-11-15
Publishing country	United States
Document type	Journal Article
ZDB-ID	1491859-6
ISSN	1094-4087 ; 1094-4087
ISSN (online)	1094-4087
ISSN	1094-4087
DOI	10.1364/OE.27.001497
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