Cellsens dimension system

Manufactured by Olympus

Sourced in Japan

The CellSens Dimension system is a high-performance imaging platform designed for life science research. It provides advanced imaging capabilities, including fluorescence and phase contrast microscopy, to enable detailed observation and analysis of biological samples.

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Lab products found in correlation

Uplanapo 100 1.5 oil objective, by Olympus (3 mentions) Csu w1 confocal scanner, by Yokogawa (2 mentions) Phe0023, by Thermo Fisher Scientific (1 mentions) 35 mm glass bottom dishes, by MatTek (1 mentions) Phenol red, by Sartorius (1 mentions) Csu w1, by Yokogawa (1 mentions) 35 mm glass bottomed dishes, by MatTek (1 mentions) Bromodeoxyuridine (brdu), by Merck Group (1 mentions) Rabbit anti pd 1, by Cell Signaling Technology (1 mentions) Tissue tek polar dm, by Sakura Finetek (1 mentions) Vectashield mounting medium containing 4 6 diamidino 2 phenylindole dapi, by Vector Laboratories (1 mentions) Tissue tek oct compound, by Sakura Finetek (1 mentions) Slide glasses, by Matsunami (1 mentions)

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CellSens Dimension Imaging System CellSens Dimension CellSens system CellSens

8 protocols using cellsens dimension system

Quantifying CD9-positive Cell Fractions

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Five random fields were imaged per slide for smear preparation of isolated CD9-positive cell fraction using a fluorescence microscope with a 40-fold objective
lens. The number of CD9-positive cells and the total number of DAPI-stained cells per unit area (157.5 × 210 μm²) were counted using cellSens
Dimension System (Olympus). Observations were performed in triplicate for each experimental group.

HORIGUCHI K., YOSHIDA S., TSUKADA T., NAKAKURA T., FUJIWARA K., HASEGAWA R., TAKIGAMI S, & OHSAKO S. (2020). Expression and functions of cluster of differentiation 9 and 81 in rat mammary epithelial cells. The Journal of Reproduction and Development, 66(6), 515-522.

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Isolation and Characterization of Rat CD9+ Cells

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Anterior lobes of male Wistar rats were dissected, and the cells were dispersed as described previously^{35 (link)}. Dispersed cells were counted using a haemocytometer and separated using a Universal Mouse pluriBeads kit (pluriSelect, San Diego, CA, USA) as described in our previous paper^{19 (link)} with a monoclonal anti-rat CD9 antibody (BD Biosciences). CD9-positive and -negative cells were processed for smear preparation, qPCR, cultivation, or immunocytochemistry as described above. After immunocytochemistry, using a 40× objective lens, 10 random fields (157.5 × 210 mm rectangle) were captured per cell culture chamber well containing isolated CD9-positive cells. The populations of CD9-positive cells and CD9/S100β9/SOX2-positive cells were counted using the cellSens Dimension system (Olympus). Three individual experiments were carried out for cell counting.

Horiguchi K., Fujiwara K., Yoshida S., Nakakura T., Arae K., Tsukada T., Hasegawa R., Takigami S., Ohsako S., Yashiro T., Kato T, & Kato Y. (2018). Isolation and characterisation of CD9-positive pituitary adult stem/progenitor cells in rats. Scientific Reports, 8, 5533.

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Live-Cell Imaging of Cytoskeleton Dynamics

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For live-cell imaging, 35-mm glass-bottom dishes (MatTek Corp.) were coated with 10 µg/ml fibronectin (PHE0023; Gibco) in PBS for ≥3 h at 37°C, washed with PBS twice, and immersed in complete DMEM without phenol red (01-053-1A; Biological Industries) before seeding of cells. For labeling actin and microtubule cytoskeleton, cells were incubated with 0.2 µM SiR-Actin (CY-SC001; Cytoskeleton) and SiR-Tubulin (CY-SC002; Cytoskeleton) for 6 h, respectively.
Time-lapse images of cells with transient transfection were acquired with the Olympus CellSens Dimension system, consisting of an Olympus SpinSR10 Ixplore spinning disk confocal microscope and a Yokogawa CSU-W1 confocal scanner. Appropriate filters, heated sample environment (37°C), controlled 5% CO₂, and UplanApo 100×/1.5 oil objective (Olympus Corp.) were used. The recording was set as every 10 min for 12 h, and one focal plane was recorded for all live-cell videos.

Fan C., Shi X., Zhao K., Wang L., Shi K., Liu Y.J., Li H., Ji B, & Jiu Y. (2022). Cell migration orchestrates migrasome formation by shaping retraction fibers. The Journal of Cell Biology, 221(4), e202109168.

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Live Cell Imaging of CAV-1 Vesicles

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For live cell imaging, 35 mm glass-bottomed dishes (MatTek Corporation, Ashland, MA, United States) were coated with 10 μg/ml fibronectin (#F2006, Sigma Corp., St. Louis, MO, United States) in PBS for at least 3 h at 37°C, washed with PBS twice and immersed in complete DMEM medium without phenol red (#01-053-1A, Biological Industries, Kibbutz Beit-Haemek, Israel) before seeding of cells. The time-lapse images of cells with transient transfection of CAV-1-mEGFP and mCherry-actin were acquired with Olympus cellSens Dimension system, consisting of an Olympus SpinSR10 Ixplore spinning disk confocal and a Yokogawa CSU-W1 confocal scanner. Appropriate filters, heated sample environment (+37°C), controlled 5% CO₂ and UplanApo 100×/1.5 Oil objective (Olympus Corporation, Tokyo, Japan) was used. The recording was set as every 1 s for 200 s and one focal plane was recorded for all live cell videos. For tracking and speed measurement of CAV-1 vesicles, the Imaris 9.2 (Bitplane, Zurich, Switzerland) “Track” module with globular-objects over time was used as in previous study (Jiu, 2018 (link)). Two micrometers estimated XY diameter, 5 μm max distance and 3 max gap size were set for analyzing.

Shi X., Wen Z., Wang Y., Liu Y.J., Shi K, & Jiu Y. (2021). Feedback-Driven Mechanisms Between Phosphorylated Caveolin-1 and Contractile Actin Assemblies Instruct Persistent Cell Migration. Frontiers in Cell and Developmental Biology, 9, 665919.

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BrdU Proliferation Assay of CD9-Positive Cells

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To visualise the proliferative activities of cells, the nucleotide analogue BrdU (Merck Millipore) was added at a concentration of 3 μg/mL to the primary culture of CD9-positive cells at a density of 1.0 × 10⁵ cells/cm² for 24 h after adding siRNAs against Cd9 mRNA. Cells were fixed in 4% paraformaldehyde in 0.025 M PB (pH 7.4) for 15 min at room temperature and were then treated with 4 M HCl in PBS for 10 min. Immunocytochemistry was performed as described above. Ten fields per well were randomly imaged using the cellSens Dimension system (Olympus) installed on a microscope (BX61, Olympus) with a 60× objective lens.

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Multiplex Immunofluorescence Staining of Lung Tissue

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Staining for two groups of combination of either CD8/CD103 or CD8/PD-1 was performed on formalin-fixed paraffin-embedded (FFPE) lung tissue slides. FFPE slides were deparaffinized in CitriSolv for 30 min and then immersed in alcohol series from 100, 95, 85, and 75% to distill H₂O for 5 min each for tissue hydration. For antigen retrieval, hydrated slides were steamed for 20 min in 1 mM EDTA. The slides were then blocked with 10% normal goat serum phosphate-buffered saline (PBS) for 30 min at room temperature (RT) and then were incubated with either rabbit anti-CD103 (Abcam) or rabbit anti–PD-1 (Cell Signaling) overnight at 4°C.8After rinsing in 0.1% PBST (PBS with Tween 20) solution, the slides were incubated with Alexa Fluor 488–conjugated goat anti-rabbit secondary Ab (Life Technologies). After rinsing with 0.1% PBST, the slides were then incubated with Alexa Fluor 647–conjugated mouse anti- CD8 (BioLegend) for 60 min at RT. After stringent washing in 0.1% PBST, slides were aired before mounting with 4’,6-diamidino-2- phenylindole for nuclei counterstain. Tissue staining for the Ab mixture was reviewed and representative images were captured in Olympus cellSens Dimension system. Fifteen representative image fields were captured for each patient for quantification purposes.

Wang Z., Wang S., Goplen N.P., Li C., Cheon I.S., Dai Q., Huang S., Shan J., Ma C., Ye Z., Xiang M., Limper A.H., Porquera E.C., Kohlmeier J.E., Kaplan M.H., Zhang N., Johnson A.J., Vassallo R, & Sun J. (2019). PD-1hi CD8+ resident memory T cells balance immunity and fibrotic sequelae. Science immunology, 4(36), eaaw1217.

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Live-cell imaging of mCherry-S. Tm infection

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Vimentin-GFP stable expression cells in 35 mm glass-bottom dishes (MatTek Corporation, P35G-1.5-14-C, US) in the density of 1 × 10⁵ cells per dish, followed by infection with mCherry tagged S. Tm at MOI of 10. Cells were rinsed with PBS and replaced with DMEM containing 50 μg/ml gentamycin after 1 h of infection. Cells were placed back in the incubator for an additional 2 h before live-cell imaging. The time-lapse images were acquired with Olympus cellSens Dimension system, consisting of an Olympus SpinSR10 Ixplore spinning disk confocal and a Yokogawa CSU-W1 confocal scanner. Appropriate filters, heated sample environment (37 °C), controlled 5% CO₂ and UplanApo 100×/1.5 oil objective (Olympus Corporation, Tokyo, Japan) were used. The recording was set as every 36 min for 18 h and one focal plane was recorded for all live cell movies.

Zhao S., Xu Q., Cui Y., Yao S., Jin S., Zhang Q., Wen Z., Ruan H., Liang X., Chao Y., Gong S., Sansonetti P., Wei K., Tang H, & Jiu Y. (2023). Salmonella effector SopB reorganizes cytoskeletal vimentin to maintain replication vacuoles for efficient infection. Nature Communications, 14, 478.

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Immunohistochemical Evaluation of IL-33 and TSLP

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Detection of IL-33 and TSLP by immunohistochemistry was performed as described elsewhere with minor modifications^{29 (link)}. Twenty-four hours after the last chitin inhalation, the trachea was cannulated with a 22-G blunt needle attached to a syringe, and 4% paraformaldehyde in 0.05 M phosphate buffer (pH 7.4) was infused. The lungs were immediately harvested and immersed in a fixative solution consisting of 4% paraformaldehyde in 0.05 M phosphate buffer (pH 7.4) at 4 °C for 20–24 h. The tissues were then immersed in 30% sucrose in 0.05 M phosphate buffer (pH 7.4) at 4 °C for more than 2 days, embedded in Tissue Tek OCT compound (Sakura Finetek Japan, Tokyo, Japan), and rapidly frozen. Frozen frontal and horizontal sections (8-μm thickness) were prepared using a cryostat (Tissue-tek Polar DM; Sakura Finetek Japan) and mounted on slide glasses (Matsunami, Osaka, Japan). Nuclei were counterstained by incubation with Vectashield Mounting Medium containing 4,6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA). Sections were scanned using a fluorescence microscope (cellSens Dimension System; Olympus, Tokyo, Japan). Each section was scanned at least three times.

Arae K., Ikutani M., Horiguchi K., Yamaguchi S., Okada Y., Sugiyama H., Orimo K., Morita H., Suto H., Okumura K., Taguchi H., Matsumoto K., Saito H., Sudo K, & Nakae S. (2021). Interleukin-33 and thymic stromal lymphopoietin, but not interleukin-25, are crucial for development of airway eosinophilia induced by chitin. Scientific Reports, 11, 5913.

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