Assessing Neuronal Viability and Outgrowth

The Hoechst/PI assay and total neurite outgrowth assessment were conducted according to a previous study (14 (link)). For the Hoechst/PI assay, Hoechst (Sigma-Aldrich; Merck KGaA) and PI (Sigma-Aldrich; Merck KGaA) were added to the medium at 37°C followed by incubation for 30 min. Images were acquired using an inverted fluorescence microscope (Leica Microsystems GmbH), and the cell apoptotic rate was calculated by counting the total cells and damaged cells. For the total neurite outgrowth assessment, the cellular morphology was observed on a fluorescence microscope (Leica Microsystems GmbH; magnification, ×200) in 5 randomly selected fields of view. The neurite outgrowth of each cell was measured by imaging software Image Pro Plus (v6.0; Media Cybernetics, Inc.), and the total neurite outgrowth per cell was calculated as follows: total length of neurite outgrowth / number of included cells.

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Zhou B., Li L., Qiu X., Wu J., Xu L, & Shao W. (2020). Long non-coding RNA ANRIL knockdown suppresses apoptosis and pro-inflammatory cytokines while enhancing neurite outgrowth via binding microRNA-125a in a cellular model of Alzheimer's disease. Molecular Medicine Reports, 22(2), 1489-1497.

Publication 2020

Hoechst inverted fluorescence microscope fluorescence microscope Image Pro Plus

Apoptotic Assay Fluorescence microscope Neurite outgrowth

Corresponding Organization :

Other organizations : Wuhan No.1 Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Cell apoptotic rate (calculated by counting the total cells and damaged cells)
Total neurite outgrowth per cell (calculated as total length of neurite outgrowth / number of included cells)

control variables

Hoechst and PI added to the medium at 37°C followed by incubation for 30 min
Cellular morphology observed on a fluorescence microscope in 5 randomly selected fields of view
Neurite outgrowth measured using imaging software Image Pro Plus (v6.0; Media Cybernetics, Inc.)

controls

None explicitly mentioned

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