Tcs sp5 confocal microscope

Manufactured by Leica

Sourced in Germany, United States, United Kingdom, Canada, Japan, Netherlands, France, Australia, Italy

The Leica TCS SP5 confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It features a modular design, allowing for customization to meet specific research requirements. The TCS SP5 is capable of capturing high-resolution, multi-dimensional images by utilizing a combination of laser sources and sophisticated optics.

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Lab products found in correlation

Las af software, by Leica (55 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (53 mentions) Alexa fluor 488, by Thermo Fisher Scientific (51 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (38 mentions) Vectashield, by Vector Laboratories (28 mentions) Bovine serum albumin (bsa), by Merck Group (25 mentions) Hoechst 33342, by Thermo Fisher Scientific (22 mentions) Triton x 100, by Merck Group (21 mentions) Las af lite software, by Leica (18 mentions) Hoechst 33342, by Merck Group (18 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (15 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (13 mentions) Paraformaldehyde (pfa), by Merck Group (13 mentions) Las af, by Leica (12 mentions) Vectashield mounting medium, by Vector Laboratories (11 mentions) Propidium iodide, by Merck Group (10 mentions) Las x software, by Leica (10 mentions) Prolong gold antifade, by Thermo Fisher Scientific (10 mentions) Prolong gold, by Thermo Fisher Scientific (9 mentions) Alexa fluor 594, by Thermo Fisher Scientific (8 mentions)

Close spelling variants of this product

Confocal TCS SP5 TCS SP5 microscope SP5 confocal microscope TCS SP5 Confocal SP5 SP5 microscope Confocal microscope

1 593 protocols using tcs sp5 confocal microscope

Immunofluorescence Imaging of Adherent Cells and Vascular Organoids

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Adherent cells: Indirect immunofluorescence on adherent cells was performed as previously described (Bellou, 2012 #68) using primary and secondary antibodies (listed in Materials and Methods Supplementary). Cell nuclei were stained using propidium iodide-PI (Sigma), samples were mounted in moviol-dabco, and images of nine fields were taken on a Leica TCS SP5 confocal microscope using HCX PL APO CS 40 × 1.25 OIL objective.
Vascular organoids/spheroids: Vascular organoids or spheroids consisting of 1,000 cells/spheroid were fixed in 3.7% paraformaldehyde for 1 h at RT, permeabilized with 0.2% Triton-X/0.9% gelatin solution for 1 h, and 0.5% Triton-X/0.9% gelatin solution for 15 min, and incubated with primary antibodies overnight at 4°C (Supplementary Table S2). Next day, the vascular organoids were washed 5x with 0.2% Triton-X and incubated with secondary antibodies for 1 h (Supplementary Table S2). After rinsing 5x with 0.2% Triton-X and incubation with Draq5 (Thermo Fisher Scientific) for 10 min, images were taken on a Leica TCS SP5 confocal microscope using HCX PL APO CS 40 × 1.25 OIL objective. At least 10 vascular organoids or spheroids were analyzed per experiment.

Markou M., Kouroupis D., Badounas F., Katsouras A., Kyrkou A., Fotsis T., Murphy C, & Bagli E. (2020). Tissue Engineering Using Vascular Organoids From Human Pluripotent Stem Cell Derived Mural Cell Phenotypes. Frontiers in Bioengineering and Biotechnology, 8, 278.

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Measuring Intracellular ROS and SOD Activity

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The cells were loaded with 10 μM H2DCF-DA (Invitrogen, Molecular Probes, USA) or 10 μM dihydroethidium (DHE, Invitrogen, Molecular Probes, USA) at 37°C for 30 minutes according to the manufacturer's instructions. After removing excess probes, the cells were kept at 37°C containing 5% CO₂. Fluorescence intensity was detected by Leica TCS SP5 confocal microscope (Leica, Germany). For each sample 10,000 events were collected. For lung tissues, add 0.1% ROS (DHE) probe about 10 edged up on the lung tissue of frozen section, after incubation for 30 minutes at 37°C, washing 2-3 times with phosphate-buffered saline (PBS) and observation by Leica TCS SP5 confocal microscope. Superoxide dismutases (SOD) activity was measured using the SOD assay kit WST (Nanjing jiancheng bioengineering institute, china). The ELISA kits of TNF-α and IL-6 were purchased from R&D Systems (Minneapolis, MN).

Chen Y., Luo G., Yuan J., Wang Y., Yang X., Wang X., Li G., Liu Z, & Zhong N. (2014). Vitamin C Mitigates Oxidative Stress and Tumor Necrosis Factor-Alpha in Severe Community-Acquired Pneumonia and LPS-Induced Macrophages. Mediators of Inflammation, 2014, 426740.

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Immunohistochemistry and TUNEL Assay for Apoptosis Analysis

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Immunohistochemistry for cleaved caspase-3 was conducted using an anti-rat cleaved caspase-3 antibody (Abcam, Cambridge, UK) and a secondary polyclonal antibody to rabbit IgG coupled with FITC (Abcam, Cambridge, UK). Fluorescence images were captured with a Leica TCS-SP5 confocal microscope (Leica, Wetzlar, Germany) and processed with Leica X software (Version 3.7.4). Apoptosis in infarcted brain tissue was analyzed using an In Situ Cell Death Detection Kit (Roche, Basel, Switzerland) according to the manufacturer’s instructions. Fluorescence from TUNEL stained signals was captured with a Leica TCS-SP5 confocal microscope and processed with Leica X software (Version 3.7.4). Neurological deficits were evaluated as previously described [11 (link)].

Chung K., Ullah I., Yi Y., Kang E., Yun G., Heo S., Kim M., Chung S.E., Park S., Lim J., Lee M., Rhim T, & Lee S.K. (2024). Intranasal Delivery of Anti-Apoptotic siRNA Complexed with Fas-Signaling Blocking Peptides Attenuates Cellular Apoptosis in Brain Ischemia. Pharmaceutics, 16(2), 290.

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Visualizing StMYB44 Subcellular Localization

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Agrobacterium cells containing 35S:StMYB44-GFP and 35S:GFP plasmids, respectively, were infiltrated into 4-week-old Nicotiana benthamiana leaves. Three days after infiltration, the leaves were detached and green fluorescent protein (GFP) signals were examined under a Leica TCS-SP5 confocal microscope (Leica Microsystems Exton, PA, USA) with excitation wavelength at 488 nm and emission wavelength at 500–520 nm.
Six-day-old transgenic Arabidopsis seedlings expressing the 35S:StMYB44-GFP and 35S:GFP transgenes were used to study subcellular localization. Nuclei of root cells were stained with DAPI solution at 10 μg ml^–1 (w/v) for 10 min, and then washed three times with water. Transgenic Arabidopsis seedlings expressing 35S:GFP were used as the control. GFP and DAPI signals were examined using a Leica TCS-SP5 confocal microscope with excitation wavelengths 488 nm for GFP and 405 nm for DAPI (Zhou et al., 2011 (link)).

Zhou X., Zha M., Huang J., Li L., Imran M, & Zhang C. (2017). StMYB44 negatively regulates phosphate transport by suppressing expression of PHOSPHATE1 in potato. Journal of Experimental Botany, 68(5), 1265-1281.

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Immunostaining of HCGR in Cells

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Immunostaining was carried out using recommended protocol and imaging was performed on a Leica TCS-SP5 confocal microscope. Briefly, cells were fixed with 4% paraformaldehyde min and then blocked in 5% goat serum (Biotech Well), followed by primary antibody incubation overnight at 4°C. The primary antibody was monoclonal mouse anti-HCGR (1:500, Abcam). Following primary antibody incubation and washes, cells were incubated with Cy3-conjugated goat anti-mouse IgG (Jackson ImmunoResearch) for 30 min in the dark at room temperature. DAPI (Sigma-Aldrich) was used for nuclear identification. Cells were observed using a Leica TCS-SP5 confocal microscope after staining.

Cong Q., Lin L., Qi B., Xu C, & Zhang X. (2021). Human Chorionic Gonadotropin Polypeptide Nanoparticle Drug Delivery System Improves Methotrexate Efficacy in Gestational Trophoblastic Neoplasia in vitro. Cancer Management and Research, 13, 1699-1708.

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Visualizing Root and Shoot Apical Meristems

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Roots of ARF transcriptional reporter lines were imaged at 5 days after germination. Plant cell walls were visualized by staining with 15 μg/ml propidium iodide solution. Roots were examined using a TCS-SP5 confocal microscope (Leica) with excitation at 514 nm and emission at 526-560 nm for mVenus and 605-745 nm for propidium iodide.
For analysis of shoot apical meristems, bolted shoots were dissected under a stereomicroscope and transferred to an Apex Culture Medium (1/2 MS medium supplemented with 1% sucrose, 0.8% agarose, 1x vitamin solution (myo-Inositol 100 mg/L, nicotinic acid 1 mg/L, pyridoxine hydrochloride 1 mg/L, thiamine hydrochloride 10 mg/L, glycine 2 mg/L)), for overnight incubation. Before microscopy cell walls were stained with 100 μg/ml propidium iodide solution. The shoot apices were then examined using a TCS-SP5 confocal microscope (Leica) with excitation at 514 nm and emission at 526-560 nm for mVenus and 605-745 nm for propidium iodide.

Truskina J., Han J., Chrysanthou E., Galvan-Ampudia C.S., Lainé S., Brunoud G., Macé J., Bellows S., Legrand J., Bågman A.M., Smit M.E., Smetana O., Stigliani A., Porco S., Bennett M.J., Mähönen A.P., Parcy F., Farcot E., Roudier F., Brady S.M., Bishopp A, & Vernoux T. (2021). A network of transcriptional repressors modulates auxin responses. Nature, 589(7840).

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FRAP Analysis of Par3N Droplets

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The in vitro FRAP analysis of iFluor488-Par3N droplets was carried out in a 1:1 mixture of Par3N and Par6β (25 µM) at room temperature. The 488 signal was bleached using a 488-nm laser beam with a Leica TCS SP5 confocal microscope.
COS7 cells were cultured in glass bottom dishes and transfected as described above. FRAP assay was performed on a Leica TCS SP5 confocal microscope. Puncta with diameters ~1.0 µm were assayed. GFP signal was bleached using a 488-nm laser beam. The fluorescence intensity difference between pre-bleaching and at time 0 (the time point right after photobleaching pulse) was normalized to 100%. The experimental control is to quantify fluorescence intensities of similar puncta/cytoplasm regions without photobleaching.

Liu Z., Yang Y., Gu A., Xu J., Mao Y., Lu H., Hu W., Lei Q.Y., Li Z., Zhang M., Cai Y, & Wen W. (2020). Par complex cluster formation mediated by phase separation. Nature Communications, 11, 2266.

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Quantitative Confocal Imaging Analysis

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IF images were acquired on a Leica TCS SP5 confocal microscope, using a × 40/1.25N.A. objective (Leica HCX PL APO lambda blue × 40/1.25 oil UV). Several fields of views were acquired with tiling scan function in order to get an area of 1,000 × 1,000 μm². Image analysis was performed measuring total fluorescence of TF both in nuclear and cytoplasmic region, with a customized pipeline in the CellProfiler software66 (link). Images for tissue samples stained for Vimentin/E-cadherin were acquired on a Leica TCS SP5 confocal microscope using × 10/1.25 oil.

Revandkar A., Perciato M.L., Toso A., Alajati A., Chen J., Gerber H., Dimitrov M., Rinaldi A., Delaleu N., Pasquini E., D'Antuono R., Pinton S., Losa M., Gnetti L., Arribas A., Fraering P., Bertoni F., Nepveu A, & Alimonti A. (2016). Inhibition of Notch pathway arrests PTEN-deficient advanced prostate cancer by triggering p27-driven cellular senescence. Nature Communications, 7, 13719.

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Adipocyte Nuclei and Diameter Analysis

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Adipose tissue was collected, embedded in paraffin, and stained as previously described (Holtrup, et al. 2017 (link); Jeffery et al. 2015 (link); Jeffery et al. 2016 (link); Sebo and Rodeheffer 2021 ). For adipocyte nuclei analysis, 20–30 images for every tissue section were acquired at 40X with a Leica TCS SP5 confocal microscope. Quantification of BrdU in adipocyte nuclei was done as previously described (Jeffery et al. 2015 (link)). At least 50 adipocyte nuclei were scored for each animal.
For adipocyte diameter measurements, the area of each adipocyte (in square pixels) was measured using Cell Profiler. The diameter of each adipocyte was calculated using the measured area, assuming each adipocyte is a perfect circle. At least 200 adipocytes were measured for each animal.
For whole mount microscopy, tissues were dissected and cut into ~1.5×1.5 cm pieces. Samples were subsequently mounted onto microscope slides with Fluoromount-G (SouthernBiotech, 0100–01) and imaged at 20X with a Leica TCS SP5 confocal microscope.

Saavedra-Peña R.D., Taylor N, & Rodeheffer M.S. (2022). Insights of the role of estrogen in obesity from two models of ERα deletion. Journal of molecular endocrinology, 68(4), 179-194.

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Localization and Functions of FgNTH and FgATH in Fusarium

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Localization of FgNTH and FgATH was determined with strains FgNTH‐GFP and FgATH‐GFP grown on YEPD (yeast extract 3 g/L, peptone 10 g/L, glucose 20 g/L) (for hyphae) and CMC (for conidia); FgNTH‐GFP and FgATH‐GFP were examined with a Leica TCS SP5 confocal microscope. The hyphae and conidia of strains FgNTH‐GFP and FgATH‐GFP were stained with 4,6‐diamidino‐2‐phenylindole (DAPI) to observe nuclei. The hyphae of strains FgNTH‐GFP, FgATH‐GFP, ΔFgNTH FgATH‐GFP, ΔFgATH FgNTH‐GFP, and ΔFgNTH FgPK‐GFP were stained with 7‐amino‐4‐chloro‐methylcoumarin (CMAC) to observe vacuoles.
To detect the effect of FgNTH and FgATH on Fusarium toxisomes, the strains ΔFgNTH FgTRI1‐GFP, ΔFgNTH FgTRI5‐GFP, ΔFgATH FgTRI1‐GFP, and ΔFgATH FgTRI5‐GFP were cultured in GYEP medium for 3 days, and then the expression and location of FgTRI1 and FgTRI5 in their hyphae were assessed with a Leica TCS SP5 confocal microscope.

Bian C., Duan Y., Xiu Q., Wang J., Tao X, & Zhou M. (2021). Mechanism of validamycin A inhibiting DON biosynthesis and synergizing with DMI fungicides against Fusarium graminearum. Molecular Plant Pathology, 22(7), 769-785.

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