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Tcs sp5 2 inverted confocal microscope

Manufactured by Leica
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The Leica TCS SP5-II is an inverted confocal microscope. It is designed to provide high-resolution imaging of samples using laser-scanning technology. The instrument is capable of capturing detailed images of cellular and subcellular structures.

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Lab products found in correlation

Human ifn γ, by Thermo Fisher Scientific (2 mentions) Annexin 5 fitc apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Annexin 5 fitc pi, by Thermo Fisher Scientific (1 mentions) Apodetect kit, by Thermo Fisher Scientific (1 mentions) Hoechst 33258, by Thermo Fisher Scientific (1 mentions) Ab28448, by Abcam (1 mentions) D9542, by Merck Group (1 mentions) S3023, by Agilent Technologies (1 mentions) A0024, by Abcam (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

TCS SP5 inverted confocal microscope TCS SP5 II inverted microscope TCS SP5 II confocal microscope TCS SP5 inverted microscope TCS SP5 II confocal SP5 inverted confocal microscope TCS SP5 II microscope SP5 II confocal microscope Confocal TCS SP5 Inverted Confocal SP5

4 protocols using tcs sp5 2 inverted confocal microscope

1

Generating hES-Derived Neuronal Cultures

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hES derived DA neuronal cultures were obtained as described28 (link). Briefly, H9 human ES cells were subjected to dual SMAD-inhibition followed by exposure to sonic hedgehog, purmorphamine, FGF8 and CHIR99021. Neurons were passaged on day 20, replated on glass-bottom dishes on day 30 at a density of 200.000 cells per 20 μl droplet, and then matured in Neurobasal media with the addition of B27 supplement, GDNF, BDNF, ascorbic acid, cAMP, TGFbeta and DAPT. hES derived spinal motor neuron (MN) cultures expressing EYFP under control of the human synapsin promoter were generated by exposure to dual SMAD-inhibition followed by caudalization with retinoid acid and ventralization through the hedgehog pathway. MN aggregates were replated on day 20 and matured in Neurobasal media with the addition of B27 supplement, GDNF, BDNF, ascorbic acid, cAMP and DAPT.
Cultures were treated with human IFN-γ (eBioscience) at a non-toxic concentration for hES (100 ng/ml, 72h). Cultures were then incubated with the MHC-I antibody prior to fixation in 4% paraformaldehyde and double immunofluorescence for TH and MHC-I as above. Micrographs were acquired using a Leica TCS SP5-II inverted confocal microscope.
Cebrián C., Zucca F.A., Mauri P., Steinbeck J.A., Studer L., Scherzer C.R., Kanter E., Budhu S., Mandelbaum J., Vonsattel J.P., Zecca L., Loike J.D, & Sulzer D. (2014). MHC-I expression renders catecholaminergic neurons susceptible to T-cell-mediated degeneration. Nature communications, 5, 3633.
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2

Annexin V-FITC Apoptosis Detection

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The cell surface exposure of phosphatidylserine on the plasma membrane of cells were assessed using Annexin V-FITC Apoptosis Detection Kit (Life-Technologies Apo-Detect Kit). Briefly, cells were harvested on a coverslip sited in a 24-well plate at a density of 104 cells per well. After adhesion, cells were treated with phenformin at 0, 1 and 10 mM. After 24 hours of treatment cells were washed with PBS and supernatants were conserved for cyto-spin. Coverslips were incubated with a mix of Annexin V-FITC, PI and Hoechst 33258 (Thermofisher) in the dark for 10 min at room temperature. The same procedure was reserved for cells recovered in the supernatants after centrifugation. Cells were fixed with PFA 4% for 10 minutes. After washing with PBS, coverslips were mounted with Dako and the fluorescence images were obtained with a Leica TCS-SP5 II inverted confocal microscope.
Coperchini F., Croce L., Denegri M., Awwad O., Ngnitejeu S.T., Magri F., Chiovato L, & Rotondi M. (2019). The anti-cancer effects of phenformin in thyroid cancer cell lines and in normal thyrocytes. Oncotarget, 10(60), 6432-6443.
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3

Immunofluorescence Staining of Neural Cells

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For immunofluorescence staining, cells were fixed with 4% formaldehyde in PBS solution for 15 min, permeabilized with 1% Triton X-100 in PBS solution for 15 min and blocked with 2% bovine serum albumin in PBS solution for 1 h. Thereafter, cells were incubated with the following primary antibodies overnight at 4 °C: anti-MAP 2 (1:500; Sigma, M1406), anti-TUJ1 (1:500; Covance, MRB-435P), anti-TAU (1:200; Dako, A0024) and anti-CTIP2 (1:200; Abcam, ab28448). Primary antibodies were detected using secondary antibodies conjugated to Alexa Fluor 488 (1:500; Molecular Probes) and Alexa Fluor 594 (1:500; Molecular Probes). Cells were washed in PBS and nuclei counterstained with DAPI (Sigma-Aldrich, D9542). Finally, coverslips of growing cells were transferred onto glass slides with mounting medium (Dako, S3023) and imaged immediately using sequential line scanning with a Leica TCS SP5 II inverted confocal microscope.
Aldana B.I., Zhang Y., Jensen P., Chandrasekaran A., Christensen S.K., Nielsen T.T., Nielsen J.E., Hyttel P., Larsen M.R., Waagepetersen H.S, & Freude K.K. (2020). Glutamate-glutamine homeostasis is perturbed in neurons and astrocytes derived from patient iPSC models of frontotemporal dementia. Molecular Brain, 13, 125.
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4

Generating hES-Derived Neuronal Cultures

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
hES derived DA neuronal cultures were obtained as described28 (link). Briefly, H9 human ES cells were subjected to dual SMAD-inhibition followed by exposure to sonic hedgehog, purmorphamine, FGF8 and CHIR99021. Neurons were passaged on day 20, replated on glass-bottom dishes on day 30 at a density of 200.000 cells per 20 μl droplet, and then matured in Neurobasal media with the addition of B27 supplement, GDNF, BDNF, ascorbic acid, cAMP, TGFbeta and DAPT. hES derived spinal motor neuron (MN) cultures expressing EYFP under control of the human synapsin promoter were generated by exposure to dual SMAD-inhibition followed by caudalization with retinoid acid and ventralization through the hedgehog pathway. MN aggregates were replated on day 20 and matured in Neurobasal media with the addition of B27 supplement, GDNF, BDNF, ascorbic acid, cAMP and DAPT.
Cultures were treated with human IFN-γ (eBioscience) at a non-toxic concentration for hES (100 ng/ml, 72h). Cultures were then incubated with the MHC-I antibody prior to fixation in 4% paraformaldehyde and double immunofluorescence for TH and MHC-I as above. Micrographs were acquired using a Leica TCS SP5-II inverted confocal microscope.
Cebrián C., Zucca F.A., Mauri P., Steinbeck J.A., Studer L., Scherzer C.R., Kanter E., Budhu S., Mandelbaum J., Vonsattel J.P., Zecca L., Loike J.D, & Sulzer D. (2014). MHC-I expression renders catecholaminergic neurons susceptible to T-cell-mediated degeneration. Nature communications, 5, 3633.
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