Dmi 6000b cs

Manufactured by Leica camera

Sourced in Germany

The DMI 6000B CS is a compact and versatile inverted microscope designed for a wide range of applications. It features a modular design that allows for the integration of various accessories and configurations to suit different research needs. The microscope's core function is to provide high-quality imaging and observation capabilities for a variety of samples, including cell cultures, tissue sections, and other biological specimens.

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Lab products found in correlation

Fluorescein phalloidin, by Thermo Fisher Scientific (1 mentions) Saponin, by Merck Group (1 mentions) Tcs sp5 aobs confocal microscope, by Leica camera (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Gtx113286, by GeneTex (1 mentions) Gtx101279, by GeneTex (1 mentions) Sab2101014, by Merck Group (1 mentions) Affinipure goat anti mouse fitc, by Jackson ImmunoResearch (1 mentions) Affinipure goat anti rabbit rhodamine, by Jackson ImmunoResearch (1 mentions) Vectashield antifade mounting medium h 1000, by Vector Laboratories (1 mentions) Spermidine, by Merck Group (1 mentions) Dmi4000b, by Leica camera (1 mentions) Anti stat3 monoclonal antibody, by Cell Signaling Technology (1 mentions)

7 protocols using dmi 6000b cs

Confocal Microscopy of Actin Cytoskeleton

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For confocal microscopy (Leica,
DMI6000B-CS), after 24 h exposure of the materials and/or the indirect
method as described above, MEM was removed, and cells washed thrice
with PBS 1×, before being fixed with 4% formalin solution neutral
buffered for 20 min, permeabilised with 0.1% saponin (Sigma-Aldrich)
for 5 min before staining the actin filaments using fluorescein-phalloidin
(488 nm, Invitrogen Corp.), and the nucleus with DAPI (364 nm, Sigma-Aldrich).
Images of the structure and organization of actin filaments (i.e.,
the cytoskeleton) and cell morphology/phenotype were acquired with
a Leica TCS-SP5 AOBS confocal microscope. For multicolored microscopy,
samples were excited with 364 and 488 nm laser lines, and images were
captured by multitracking to avoid bleed-through between the fluorophores.

Pereira R.F., Zehbe K., Günter C., dos Santos T., Nunes S.C., Paz F.A., Silva M.M., Granja P.L., Taubert A, & de Zea Bermudez V. (2018). Ionic Liquid-Assisted Synthesis of Mesoporous Silk Fibroin/Silica Hybrids for Biomedical Applications. ACS Omega, 3(9), 10811-10822.

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Immunofluorescent Staining of Cytoskeletal Proteins

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Cells were fixed with 4% paraformaldehyde in PBS for 15 min at room temperature and permeabilized with 0.1% Triton X-100 in PBS for 5 min at room temperature. Samples were blocked for 30 min in PBS with 2% bovine serum albumin (BSA). Primary antibodies against β-tubulin (GTX101279, GeneTex, Irvine, CA, USA), γ-tubulin (GTX113286, GeneTex, Irvine, CA, USA), acetyl-α-tubulin (GTX16292, GeneTex, Irvine, CA, USA), and HAS3 (SAB2101014, Sigma-Aldrich, St. Louis, MO, USA) were diluted 1:100 in PBS with 1% BSA, and then incubated with the cells for 2 h at room temperature. AffiniPure goat anti-mouse FITC (15-095-003, Jackson ImmunoResearch, West Grove, PA, USA) and AffiniPure goat anti-rabbit rhodamine (111-025-144, Jackson ImmunoResearch, West Grove, PA, USA) secondary antibodies were diluted 1:50. Slides were incubated with secondary antibodies for 1 h at room temperature. Samples were mounted with VECTASHIELD Antifade Mounting Medium (H-1000, Vector Laboratories, Burlingame, CA, USA) and imaged via confocal microscopy (DMI 6000B CS, Leica, Wetzlar, Germany).

Lee W.J., Tu S.H., Cheng T.C., Lin J.H., Sheu M.T., Kuo C.C., Changou C.A., Wu C.H., Chang H.W., Chang H.L., Chen L.C, & Ho Y.S. (2021). Type-3 Hyaluronan Synthase Attenuates Tumor Cells Invasion in Human Mammary Parenchymal Tissues. Molecules, 26(21), 6548.

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Two-photon Lifetime Imaging of AmCyan and EYFP

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FLIM was carried out using confocal microscopy (DMI 6000B CS, Leica, Wetzlar, Germany). The AmCyan fluorescence lifetime was measured upon 2-photon excitation at 405 nm. A 495 nm long pass filter was used to separate the blue and green fluorescence channels. AmCyan emission was detected with a blue filter (BP 420) in one channel, and enhanced yellow fluorescent protein (EYFP) emission was detected with a green filter (BP 500). Images were acquired using a 63× oil objective. Images with a resolution of 512 × 512 pixels were collected with a minimum of 1000 counts per pixel, which requires integrating at least 30 frames with a pixel dwell time of 25.6 us/pixel. The temperature was set to 37 °C throughout the experiment conducted in a 5% CO₂ atmosphere.

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Subcellular Localization of BoALMT1 in Onion Cells

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The subcellular localization of BoALMT1 was determined in onion (Allium cepa) epidermal cells. We constructed a vector as 35S:BoALMT1::GFP. The coding region of BoALMT1 was subcloned into the expression vector pCAMBIA1302 using primers: 5′-CATGCCATGGTAATGGAGAAACTGAGAGAGATAGTG-3′ (forward) and 5′-GGACTAGTAATCTGAAGTATACGAACACCC-3′ (reverse). We transferred the chimera by particle bombardment. The gold particles (1 μm, 1.5 mg) were coated with 5 μg of plasmid DNA in a solution of 2.5 M CaCl₂ and 0.1 M spermidine (Sigma). We bombarded the epidermal onion peels at a helium pressure of 25–30 Mpa (Bio-rad, U.S.), and then incubated the tissue in MS medium at room temperature in the dark for 24 h. Confocal laser scanning microscopy (Leica DMI 6000B-CS, Germany) with a 488 nm excitation wavelength was used to detect the GFP fluorescence. We induced cell plasmolysis by adding 0.8 M mannitol for 3–5 min.

Zhang L., Wu X.X., Wang J., Qi C., Wang X., Wang G., Li M., Li X, & Guo Y.D. (2018). BoALMT1, an Al-Induced Malate Transporter in Cabbage, Enhances Aluminum Tolerance in Arabidopsis thaliana. Frontiers in Plant Science, 8, 2156.

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Optimization of Plasmid Transfection Efficiency

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Briefly, 1.5 × 10⁵ cells were washed twice with PBS and mixed with pSUPER-Sh-HAS3, pSUPER-Sc-HAS3, HAS3-EYFP-pcDNA3.1, or α-tubulin-AmCyan (10 μg) plasmid. Two pulses were applied for a duration of 20 milliseconds under a fixed voltage of 1.2 kV on an MP-100 pipette-type microporator (Digital Bio, Seoul, Korea). The transfection efficiency was determined by fluorescence microscopy (DMI 4000B, Wetzlar, Germany) and confocal microscopy (DMI 6000B CS, Leica, Wetzlar, Germany). The primer sequences were designed as follows: HAS3-EYFP-pcDNA3.1 sense sequence: 5′-ATTAAGCTTATGCCGGTGCAGCTGACGACA-3′, antisense sequence: 5′-TGAGAATTCCACACCTCAGCAAAAGCCA-3′ and α-tubulin-AmCyan sense sequence: 5′-ATAGCTAGCATGCGTGAGTGCATCTCCATC-3′ and antisense sequence: 5′-GAAACCGGTGTATTCCTCTCCTTCTTCCTC-3′.

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Quantifying Cell Binding and Internalization of RNA Nanoparticles

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For the cell binding assay, 3WJ-B-α9-apt-Alexa and 3WJ-C-α9-apt-Alexa RNA nanoparticles were incubated with MDA-MB-231 cells at 37°C for 1 h before analysis by flow cytometry. The contents of nanoparticles in live cells were detected with excitation at 640 nm and emission at 680 nm. MDA-MB-231 cells were seeded in a 3-cm glass Petri dish overnight to study internalization. Then, 3WJ-B-α9-apt-Alexa and 3WJ-B-α9-apt-Alexa RNA nanoparticles were added directly to cells in the live-cell system with a real-time laser confocal microscope (DMI 6000B CS, Leica, Wetzlar, Germany). Hochest 33342 was added for cell nucleus staining. The temperature was set to 37°C throughout the experiment in a 5% CO₂ atmosphere, following the manufacturer’s instructions. Live-cell images were acquired synchronously using a 63× oil objective to observe internalization and cell entry with excitation at 640 nm and emission at 680 nm. Consecutive images were concatenated into video clips using PowerDirector software and uploaded to YouTube.

Liao Y.C., Cheng T.C., Tu S.H., Chang J., Guo P., Chen L.C, & Ho Y.S. (2023). Tumor targeting and therapeutic assessments of RNA nanoparticles carrying α9-nAChR aptamer and anti-miR-21 in triple-negative breast cancers. Molecular Therapy. Nucleic Acids, 33, 351-366.

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Dissecting STAT3 Activation in IL-17-Stimulated HMECs

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HMECs were plated into 6-well plates and pre-treated with vehicle (0.1% DMSO), FLLL31 (10 μM), or AG490 (10 μM) for 30 minutes. The cells were then stimulated with IL-17 (50 ng/ml) for 1 hour. Treated cells were washed with ice-cold PBS, fixed with cold methanol, and cultured in blocking buffer for 30 minutes. Following blocking, the cells were washed with PBS and cultured with an anti-STAT3 monoclonal antibody (Cell Signaling Technology) overnight at 4°C. After washing, the cells were labeled with a FITC-conjugated anti-mouse secondary antibody, stained with DAPI, and viewed with a scanning confocal imaging system. Six random fields in each well (magnification × 60, Leica DMI 6000B-CS) were chosen for analysis.
To examine the nuclear translocation of STAT3 and its dominant-negative mutants after IL-17 treatment, the GFP-fused wild type and Y705F mutants of STAT3 were transfected for 24 hours into HMECs with Lipofectin according to the manufacturer's instructions. Then HMECs were treated with IL-17 (50 ng/ ml) for 2 hours. The cells were viewed by fluorescence confocal microscopy and six random fields in each well were counted.

Yuan S., Zhang S., Zhuang Y., Zhang H., Bai J, & Hou Q. (2015). Interleukin-17 Stimulates STAT3-Mediated Endothelial Cell Activation for Neutrophil Recruitment. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 36(6).

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