Lsm 510 meta multiphoton confocal microscope

Manufactured by Zeiss

The LSM 510 Meta Multiphoton Confocal Microscope is a laboratory equipment designed for high-resolution imaging of biological samples. It combines the principles of multiphoton excitation and confocal microscopy to provide enhanced optical sectioning and reduced phototoxicity. The system is equipped with multiple laser sources and detectors to enable simultaneous acquisition of multiple fluorescent signals.

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Lab products found in correlation

E cadherin, by Cell Signaling Technology (2 mentions) Vimentin, by Cell Signaling Technology (2 mentions) Vectashield antifade mounting medium with dapi, by Vector Laboratories (2 mentions) Bodipy 493 503, by Thermo Fisher Scientific (1 mentions) Dsred2 mito, by Takara Bio (1 mentions) Dsred2 er, by Takara Bio (1 mentions) N cadherin, by Merck Group (1 mentions) Oct3 4, by BD (1 mentions) Twist1, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

LSM 510 Meta NLO multiphoton confocal microscope Zeiss LSM 510 Meta Multiphoton microscope LSM 510 multiphoton confocal microscope LSM 510 META confocal microscope LSM 510 META confocal LSM 510 multiphoton microscope LSM 510 META microscope 510 Meta confocal microscope LSM 510 confocal microscope LSM 510 META

5 protocols using lsm 510 meta multiphoton confocal microscope

Osteogenic Differentiation of ASCs in Hydrogels

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Similar to the adipogenic differentiation study, osteogenic differentiation was probed in the MCS+DAT, MCS+DTB, and MCS hydrogels cultured in osteogenic medium or proliferation medium as a non-induced control. For the ALP assay, P3 ASCs seeded at a density of 5,000 cells/cm² on laminin-coated (Sigma, Cat. # L2020; 1.6 μg/cm²) TCP and cultured in both media formulations were included as additional controls.
ALP enzyme activity was measured at 7 and 14 d post-induction of differentiation (n = 3 samples per group/trial, N = 3 trials with different ASC donors), following published methods (Yu et al., 2017 (link)). For each cell donor, the data was normalized to the TCP group at 7 d under non-induced conditions for comparative purposes. In addition, matrix mineralization was qualitatively assessed in the hydrogel groups at 28 d post-induction using the OsteoImage^TM (Lonza, Germany) kit (n = 3 samples per group/trial, N = 2 trials with different ASC donors) as per the manufacturer's protocol. As a negative control, cell-free hydrogels without ASCs were also prepared, cultured, and imaged using a Zeiss Multiphoton LSM 510 META confocal microscope at 25X magnification.

Shridhar A., Amsden B.G., Gillies E.R, & Flynn L.E. (2019). Investigating the Effects of Tissue-Specific Extracellular Matrix on the Adipogenic and Osteogenic Differentiation of Human Adipose-Derived Stromal Cells Within Composite Hydrogel Scaffolds. Frontiers in Bioengineering and Biotechnology, 7, 402.

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Adipogenic Differentiation of ASCs in ECM Hydrogels

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In vitro studies were performed to probe the effects of incorporating the tissue-specific ECM within the MCS hydrogels on the adipogenic differentiation of encapsulated ASCs at 14-days post-induction in adipogenic differentiation medium. Hydrogels maintained in proliferation medium were included as non-induced (NI) controls. As additional controls for the GPDH assay, 12-well tissue culture polystyrene (TCP) plates were seeded with P3 ASCs at a density of 50,000 cells/cm² and cultured in both media formulations.
To quantitatively assess differentiation, intracellular GPDH enzyme activity was measured using a GPDH Enzyme Activity Measurement Kit (Kamiya Biomedical Corporation, Cat. # KT-010, Seattle, WA, USA) (n = 3 samples per group/trial, N = 3 trials with different ASC donors) as per previously-published methods (Cheung et al., 2014 (link); Brown et al., 2015 (link)). For each donor, the data was normalized to the non-induced TCP group for comparative purposes. In addition, Bodipy® 493/503 staining (Thermo Scientific) was performed to visualize intracellular lipid accumulation in the hydrogels (n = 3 samples per group/trial, N = 3 trials with different ASC donors) following the manufacturer's protocol. The hydrogels were imaged using a Zeiss Multiphoton LSM 510 META confocal microscope, with images obtained at 25X magnification.

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Immunolocalization of STING and TBK1 in HeLa Cells

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HeLa cells were seeded on coverslips in 6-well plates the day before transfection and cultured for 24 h. Plasmids of interest were transfected for 36 h before cells were fixed with 4% paraformaldehyde. For immunostaining, cells were first blocked with 5% BSA for 1 h followed by 5-min permeabilization with 0.2% Triton X-100. Cells were then incubated with primary antibody (1:200; mouse anti-HA for STING-β and mouse anti-FLAG for TBK1) overnight at 4°C. After washing with 1× PBS for three times, goat anti-mouse IgG conjugated to TRITC (1:200; Zymed) was added and incubated for 1 h. After washing with 1× PBS for three times, the coverslips were mounted onto the slides for observation using Carl Zeiss LSM 510 META Multiphoton Confocal Microscope. DsRed2-ER and DsRed2-Mito (Clontech) served as ER and mitochondrial markers.

Wang P.H., Fung S.Y., Gao W.W., Deng J.J., Cheng Y., Chaudhary V., Yuen K.S., Ho T.H., Chan C.P., Zhang Y., Kok K.H., Yang W., Chan C.P, & Jin D.Y. (2018). A novel transcript isoform of STING that sequesters cGAMP and dominantly inhibits innate nucleic acid sensing. Nucleic Acids Research, 46(8), 4054-4071.

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Quantification of Epithelial-Mesenchymal Transition

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Cells grown on glass coverslips were treated with various primary antibodies as reported [6 (link)]: E-Cadherin (Cell Signalling, 1:500), Vimentin (Cell Signalling, 1:500), and N-Cadherin (Sigma, 1:500). The cells were then incubated with fluorochrome-conjugated secondary antibodies (Biotum, Cedarlane, Burlington, ON, CA) at the following dilutions: Goat Anti-Rabbit 594 (1:500), Goat Anti-Rabbit 488 (1:500) and Goat Anti-Mouse 488 (1:500). Vectashield anti-fade mounting medium with DAPI (Vector-labs, Burlignton, ON, CA) was used to mount the slides. Fluorescent images were taken with Zeiss LSM 510 Meta Multiphoton Confocal Microscope, and fluorescence intensities calculated with ImageJ software. The raw integrated density was calculated for each cell and normalized to the cell area. Data were presented as an average of all cells.

Tordjman J., Majumder M., Amiri M., Hasan A., Hess D, & Lala P.K. (2019). Tumor suppressor role of cytoplasmic polyadenylation element binding protein 2 (CPEB2) in human mammary epithelial cells. BMC Cancer, 19, 561.

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Epithelial-Mesenchymal Transition Protein Analysis

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Cells were grown on glass coverslips to 70–80% confluency. The cells were fixed in 4% paraformaldehyde, then permeablized in 0.5% Triton-X-100, blocked with 8% BSA + 0.01% Tween 20 in PBS and incubated in primary antibody at the following dilutions in 4% BSA overnight at 4 °C: E-Cadherin and Vimentin (Cell Signalling, 1:500), TWIST1 in 1:200 ration (Santa Cruz, Dallas, TX), ALDH1A and OCT3/4 (both 1:300 dilution) from BD biosciences. The stained cells were incubated with secondary antibody (1:500, Biotium, ON). Vectashield anti-fade mounting medium with DAPI (Vector) was used to mount the slides. Immunofluorescent images were taken using the Zeiss LSM 510 Meta Multiphoton Confocal Microscope.

Majumder M., Dunn L., Liu L., Hasan A., Vincent K., Brackstone M., Hess D, & Lala P.K. (2018). COX-2 induces oncogenic micro RNA miR655 in human breast cancer. Scientific Reports, 8, 327.

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