Techniques

1. Cellular and Molecular Techniques

Cell thawing, subculturing, freezing
Primary neuronal culture
Co-culture systems (e.g., Astrocyte–Neuron)
Transfection methods (e.g., Lipofection)
qPCR (RT-qPCR)
BrdU / EdU proliferation assays
Western blot
Receptor subunit detection (e.g., GluN2A, GluN2B)
DARTS assay (Drug Affinity Responsive Target Stability)

2. Functional and Morphological Cell Assays

Trypan blue exclusion assay
Cell viability assay
ROS detection assay (e.g., DCF-DA)
Annexin V apoptosis assay
JC-1 / TMRM mitochondrial membrane potential assays
Cell cycle analysis (MUSE system)
Neurite outgrowth analysis
Sholl analysis (axon/dendrite complexity)

3. Animal Procedures and Stress Models

Blood collection: cardiac puncture, inferior vena cava
Drug administration: oral, intranasal, intraperitoneal, subcutaneous.
Sleep deprivation model
Social defeat stress model
Social isolation stress model

4. Behavioral Analysis in Animal Models

Forced Swim Test (FST)
Tail Suspension Test (TST)
Sucrose Preference Test (SPT)
Elevated Plus Maze (EPM)
Open Field Test (OFT)
Passive Avoidance Test
Morris Water Maze
Y-Maze Test
PiezoSleep Mouse Behavioral Tracking System

5. Imaging and Quantitative Analysis

Fluorescence microscopy
ImageJ-based morphometric analysis
Neurite length and branching measurement
Sholl analysis
Colocalization analysis
Densitometry for immunoblots and immunostaining

6. In Silico and Network Pharmacology

ADMET prediction
Molecular docking (AutoDock Vina)
MM-GBSA binding energy estimation
Molecular dynamics simulations
Ligand–protein interaction profiling
Database mining: TCMSP, DisGeNET, TTD
Gene/pathway enrichment (e.g., Enrichr, DAVID)
Network construction and visualization (Cytoscape, STRING)

7. Data Management and Scientific Tools

EndNote (reference management)
Excel (data handling and preprocessing)
GraphPad Prism (statistical analysis and visualization)

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