Stellaris 8 dive

Manufactured by Leica camera

Sourced in Germany, Japan

The STELLARIS 8 DIVE is a high-performance confocal laser scanning microscope designed for advanced imaging applications. It features a modular design, allowing for customization to meet specific research needs.

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Lab products found in correlation

Tcs sp5, by Leica camera (1 mentions) Tcs sp8, by Leica camera (1 mentions) Ab207434, by Abcam (1 mentions) Sc 7294, by Santa Cruz Biotechnology (1 mentions) A0460, by Beyotime (1 mentions) Bl105a, by Biosharp (1 mentions) A0468, by Beyotime (1 mentions) Fvmpe rs, by Olympus (1 mentions) Tritc dextran, by Merck Group (1 mentions) Propidium iodide, by MP Biomedicals (1 mentions) Calcein am, by BioLegend (1 mentions) Cellmask orange actin tracking stain, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Gfap antibody, by Agilent Technologies (1 mentions) Cd68 antibody, by Bio-Rad (1 mentions) Ab36362, by Abcam (1 mentions) Ma1 513, by Thermo Fisher Scientific (1 mentions) D8w5e, by Cell Signaling Technology (1 mentions) A0099, by Agilent Technologies (1 mentions) Blocking buffer, by Beyotime (1 mentions)

6 protocols using stellaris 8 dive

In Situ Liver Decellularization and Collagen Imaging

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Mouse livers were decellularized in situ by detergent (0.5% (w/v), sodium deoxycholate, 250 ml per mouse) and water (50 ml per mouse) perfusion at a pump speed of 0.2 ml min⁻¹. After the final perfusion, the livers were removed and washed overnight in PBS. For AGE-modified collagen gels, the samples were prepared as previously described. Gels were imaged 1 day after formation.
For SHG imaging, all of the samples were imaged using the Leica TCS SP5 multiphoton confocal microscope or the Leica Stellaris 8 DIVE upright confocal microscope. The excitation wavelength was tuned to 840 nm, and a 420 ± 5 nm narrow band-pass emission controlled by a slit was used for detecting the SHG signal of collagen. The images were recorded using an inverted confocal laser-scanning microscope (Leica TCS SP8) equipped with a ×20 water-immersion objective for confocal reflection imaging. An Ar⁺ laser at 488 nm was used to illuminate the sample, and the reflected light was detected with photomultiplier tube (PMT) detectors. Scans were at 1,024 × 1,024 pixels, and all of the images were taken 80–100 μm into the samples. Collagen measurements were performed using CT-Fire software (v.2.0 beta) (https://loci.wisc.edu/software/ctfire) and ImageJ v.1.53t (https://imagej-nih-gov.stanford.idm.oclc.org/ij/).

Fan W., Adebowale K., Váncza L., Li Y., Rabbi M.F., Kunimoto K., Chen D., Mozes G., Chiu D.K., Li Y., Tao J., Wei Y., Adeniji N., Brunsing R.L., Dhanasekaran R., Singhi A., Geller D., Lo S.H., Hodgson L., Engleman E.G., Charville G.W., Charu V., Monga S.P., Kim T., Wells R.G., Chaudhuri O, & Török N.J. (2024). Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver. Nature, 626(7999), 635-642.

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CBD Modulates Immune-related Transcription Factors

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Five hundred thousand B16-F10 cells were seeded in the circle microscope cover glasses and cultured overnight. CBD (Ca²⁺ (high)) at a CBD concentration of 0, 2.5, and 4 µg/mL were co-incubated with the cells for 2 h. After the removal of the medium, the cover glasses were fixed with 4% paraformaldehyde. Then, the cells were then permeabilized with Triton X-100 (0.1%, v%) for 15 min, followed by the addition of a blocking solution consisting of bovine serum albumin (BSA, 1%) and incubated at room temperature for 30 min. Next, all samples were incubated with the primary antibodies at room temperature for 12 h, followed by staining with the corresponding secondary antibodies in dark for 1 h. The primary antibodies included ATF3(ab207434, Abcam) and NFATc1(sc-7294, SANTA CRUZ). The following secondary antibodies included IgG (H + L) Fluor 555-conjugated (A0460, Beyotime) and IgG (H + L) Fluor 647-conjugated (A0468, Beyotime). Nuclei were labeled with DAPI (BL105A, Biosharp) at room temperature for 5 min. After washing with PBS thoroughly, immunofluorescence images were then acquired with a multiphoton confocal microscopy (STELLARIS 8 DIVE, Leica).

Shi J., Ma Q., Su W., Liu C., Zhang H., Liu Y., Li X., Jiang X., Ge C., Kong F., Chen Y, & Qu D. (2023). Effervescent cannabidiol solid dispersion-doped dissolving microneedles for boosted melanoma therapy via the “TRPV1-NFATc1-ATF3” pathway and tumor microenvironment engineering. Biomaterials Research, 27, 48.

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Visualizing Tumor Vascular Leakage and Drug Delivery

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In order to visualize the leakage of tumor blood vessels in vivo, LLC tumor-bearing mice were subjected to isoflurane anesthesia after being intravenously administered with 10 mg/kg of 70 kDa TRITC-dextran (25 mg/mL, Sigma–Aldrich, St. Louis, MO, USA). The skin around the tumor was carefully dissected away, followed by that the leakage of tumor blood vessels was monitored in a real-time manner under the 25 × water immersion objective of two-photon confocal microscopy (Olympus FVMPE-RS, Tokyo, Japan).
For investigating the delivery of DOX into tumor parenchyma and assessing tumor vascular perfusion ex vivo, 8 mg/kg DOX was intravenously injected into the LLC tumor-bearing mice 1 h prior to euthanasia of mice, and FITC-CD31 was intravenously injected into the tumor-bearing mice 30 min after the injection of DOX. The tumors were harvested and then imaged by using the 25 × water immersion objective of two-photon confocal microscope (Leica STELLARIS 8 DIVE, Heidelberg, Germany).

Qian C., Zhou Y., Zhang T., Dong G., Song M., Tang Y., Wei Z., Yu S., Shen Q., Chen W., Choi J.P., Yan J., Zhong C., Wan L., Li J., Wang A., Lu Y, & Zhao Y. (2024). Targeting PKM2 signaling cascade with salvianic acid A normalizes tumor blood vessels to facilitate chemotherapeutic drug delivery. Acta Pharmaceutica Sinica. B, 14(5), 2077-2096.

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3D Spheroid Viability and Cytoskeletal Imaging

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For live/dead staining, the gels were incubated with 2 μg/ml Calcein-AM (BioLegend, 425201) and 1 μg/ml Propidium Iodide (MP Biomedicals, 0219545810) in complete medium for 30 minutes at 37°C. These were imaged on a point-scanning confocal microscope (Nikon AXR). To obtain 3-D images of the spheroids for strain quantification, the gels were fixed overnight in 4% paraformaldehyde in PBS (Thermo Fisher, J19943.K2) at 4°C, then rinsed with PBS and incubated with CellMask^™ Orange Actin Tracking Stain (1:1000, Thermo Fisher, A57244) and DAPI (2 μg/ml Sigma-Aldrich, D9542) for 48 hours at 4°C. Image stacks were acquired using a multiphoton microscope (Leica Stellaris 8 DIVE).

Burchett A., Siri S., Li J., Lu X, & Datta M. (2024). Novel 3-D macrophage spheroid model reveals reciprocal regulation of immunomechanical stress and mechano-immunological response. bioRxiv.

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Comprehensive Immunohistochemical Analysis of Brain Tissue

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Brains were cut into 25 um thick sections using a microtome (Leica SM2010R, Microtome and Microscope, Leica Microsystems, Wetzlar, Germany) and stored at −20°C in an anti-freeze solution. Brain slices were stained for IIba1 antibody (1:1000, Nordic Biolabs, Wako Chemicals, Täby, Sweden, Cat. #019-19741), CD68 antibody (1:500, Bio-Rad, Hercules, CA, United States, Cat. #MCA1957), TH antibody (1:2000, Chemicon-Millipore, Burlington, MA, United States, Cat. #AB152 and #MAB358), Gephyrin antibody (1:250, Synaptic Systems, Göttingen, Germany, Cat. #147011C3), GFAP antibody (1:500, Dako, Santa Clara, CA, United States, Cat. #Z0334), in PBS with the serum (5–10%) from the animal species of the secondary antibodies and Triton-X 100. The images were acquired using Olympus Virtual Stage 120 with extended focus imaging setting (EFI) for scan images (20x and 10x magnifications), Olympus BX53 microscopes, Shinjuku, Tokyo, Japan (20x and 60x magnification), and LEICA Stellaris 8 Dive for confocal images (40x and 63x magnification). EFI setting consists of five optical sections with a 5 um increment between two consecutive optical sections. Z-stack confocal images consist of 30 optical sections, with 0.5 um increment between two consecutive optical sections, and their 3-dimensional projection was used for analyses.

Brandi E., Torres-Garcia L., Svanbergsson A., Haikal C., Liu D., Li W, & Li J.Y. (2022). Brain region-specific microglial and astrocytic activation in response to systemic lipopolysaccharides exposure. Frontiers in Aging Neuroscience, 14, 910988.

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Immunofluorescence Analysis of Intestinal Samples

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The ileal tissue was fixed for 24 h using 4% paraformaldehyde, embedded in paraffin, and subsequently sectioned. After a series of dewaxing, hydration, and antigen repair processes, tissue sections were subjected to blocking buffer (Beyotime, Nanjing, China) for 1 h and subsequently stained. The enteroids were cultured in the cell climbing slice. They were fixed for 1 h using 4% paraformaldehyde and then subjected to blocking buffer for 1 h at room temperature. Primary antibodies against AHR (1:100, MA1-513, Invitrogen, Waltham, MA, USA), IDO1 (1:1600, D8W5E, Cell Signaling Technology), and lysozyme (1:200, A0099, Dako or 1:100, ab36362, Abcam, Cambridge, UK) were used for staining for 16 h at 4 °C. After rewarming for 1 h at room temperature, the tissue sections were incubated with secondary antibodies (1:200, Yeasen, Shanghai, China) for 1 h at room temperature. Then, the sections were incubated with DAPI (Yeasen, Shanghai, China) and imaged under a fluorescence microscope. The images were captured by a Leica DMI8 and a Leica STELLARIS 8 DIVE. Images were analysed by ImageJ software (version 1.53e).

Liu L., Xu J., Xu X., Mao T., Niu W., Wu X., Lu L, & Zhou H. (2022). Intestinal Stem Cells Damaged by Deoxycholic Acid via AHR Pathway Contributes to Mucosal Barrier Dysfunction in High-Fat Feeding Mice. International Journal of Molecular Sciences, 23(24), 15578.

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