Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.

Article Snippet: SH-SY5Y neuroblastoma cells (Human) , ATCC , CRL-2266.

Techniques: Staining, Software, Concentration Assay, Comparison, Derivative Assay

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: SH-SY5Y neuroblastoma cells (Human) , ATCC , CRL-2266.

Techniques: Staining, Software, Concentration Assay, Comparison

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: SH-SY5Y neuroblastoma cells (Human) , ATCC , CRL-2266.

Techniques: Mutagenesis, Control, Stable Transfection, Staining, Software, Comparison

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.

Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

Techniques: Staining, Software, Concentration Assay, Comparison, Derivative Assay

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

Techniques: Staining, Software, Concentration Assay, Comparison

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

Techniques: Mutagenesis, Control, Stable Transfection, Staining, Software, Comparison