Developmental Biology

1. Techniques in Developmental Biology
   1. Imaging Techniques
      1. Microscopy
         1. Light microscopy
            1. Brightfield microscopy
            2. Phase contrast microscopy
            3. Differential interference contrast (DIC) microscopy
         2. Fluorescence microscopy
            1. Confocal microscopy
            2. Two-photon microscopy
            3. Total internal reflection fluorescence (TIRF) microscopy
         3. Electron microscopy
            1. Scanning electron microscopy (SEM)
            2. Transmission electron microscopy (TEM)
         4. Super-resolution microscopy
            1. Structured illumination microscopy (SIM)
            2. Stochastic optical reconstruction microscopy (STORM)
            3. Stimulated emission depletion (STED) microscopy
      2. Live Imaging of Developing Embryos
         1. Time-lapse imaging
            1. Long-term embryo culture techniques
            2. Minimizing phototoxicity and photobleaching
         2. Multiphoton microscopy
            1. Deep tissue imaging
            2. Applications in live organism studies
         3. Light sheet fluorescence microscopy
            1. Fast volumetric imaging
            2. Reduced phototoxicity for live specimens
   2. Molecular Techniques
      1. In Situ Hybridization
         1. Whole-mount in situ hybridization
         2. Fluorescent in situ hybridization (FISH)
         3. Recent advancements in high-resolution in situ hybridization
      2. RNA Sequencing
         1. Single-cell RNA sequencing
         2. Bulk RNA sequencing
         3. Techniques for capturing rare transcripts
         4. Data analysis pipelines and challenges
      3. CRISPR-based methods
         1. CRISPR-Cas9 genome editing
         2. CRISPR interference (CRISPRi) and activation (CRISPRa)
         3. Prime editing and base editing innovations
   3. Functional Assays
      1. Gene Knockdown and Knockout Studies
         1. RNA interference (RNAi)
            1. siRNA and shRNA techniques
         2. Antisense oligonucleotides
         3. CRISPR/Cas9 knockout models
      2. Reporter Gene Assays
         1. Luciferase assays
         2. Beta-galactosidase (LacZ) reporter systems
         3. Fluorescent protein reporters (e.g., GFP, RFP)
         4. Applications in real-time monitoring of gene expression
      3. Loss-of-function and gain-of-function studies
         1. Conditional mutagenesis systems
         2. Overexpression techniques
         3. Phenotypic analysis and rescue experiments
   4. Advanced Analytical Techniques
      1. Proteomics
         1. Mass spectrometry for protein identification
         2. Quantitative proteomics (SILAC, iTRAQ)
         3. Post-translational modification mapping
      2. Metabolomics
         1. Metabolic profiling during development
         2. NMR and LC-MS/MS applications
      3. Systems biology approaches
         1. Integration of multi-omics data
         2. Computational modeling of developmental processes
   5. Emerging Techniques
      1. Organoid culture and development
         1. Modeling developmental stages in vitro
         2. Applications in disease modeling
      2. Single-cell analysis
         1. Single-cell transcriptomics and epigenomics
         2. Spatial transcriptomics in developmental contexts
      3. Tissue engineering and biofabrication
         1. Scaffold-free 3D cultures
         2. Bioprinting and synthetic development platforms