Quantitative in vivo microscopy: the return from the 'omics'

Area of Science:

• Cell biology
• Biophysics
• Molecular imaging

Background:

• Recent advances in microscopy and image analysis, alongside the use of green fluorescent protein (GFP) reporters, facilitate high-throughput in vivo studies.
• These techniques provide morphological and temporal context for biochemical pathways regulating cell function.

Purpose of the Study:

• To quantify the concentration and 3D distribution of spectrally resolved GFP-tagged proteins in vivo.
• To measure the dynamics of biological processes involving these proteins using automatic segmentation and tracking.
• To enable parallel studies of multiple cell colonies under various treatments for dynamic process analysis.

Main Methods:

• High-throughput microscopy
• Image analysis
• Green fluorescent protein (GFP) tagging and spectral resolution
• Automatic segmentation and tracking algorithms

Main Results:

• Quantification of concentration and 3D distribution of multiple GFP-tagged proteins.
• Measurement of dynamics for cellular processes involving these proteins.
• Feasibility of parallel, high-throughput monitoring of drug or repressor-treated cell colonies.

Conclusions:

• This integrated approach allows for detailed in vivo analysis of cellular processes.
• It provides a powerful platform for studying the dynamics of biological pathways and the effects of perturbations.
• Enables high-throughput screening for drug discovery and understanding gene regulation.