Ultrastructural 3D Microscopy for Biomedicine: Principles, Applications, and Perspectives.

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Modern biomedical research necessitates detailed 3D analysis of biological ultrastructure.
• Conventional 2D microscopy methods are insufficient for comprehensive ultrastructural investigation.
• Developing new 3D reconstruction techniques is crucial for advancing biological studies.

Purpose of the Study:

• To review and compare various 3D microscopy techniques for ultrastructural analysis.
• To discuss the principles, applications, advantages, and limitations of different 3D imaging methods.
• To highlight advancements in electron, optical, and scanning probe microscopy for 3D reconstruction.

Main Methods:

• Serial section transmission electron microscopy (ssTEM)
• Scanning electron microscopy (SEM)-based techniques: array tomography, focused ion beam SEM (FIB-SEM), serial block-face SEM (SB-SEM)
• Super-resolution optical microscopy: stochastic optical reconstruction microscopy (STORM), stimulated emission depletion microscopy (STED)
• Scanning probe microscopy (SPM) for 3D ultrastructure
• Combination of SPM with optical techniques

Main Results:

• Detailed principles and applications of ssTEM, SEM-based methods, and super-resolution optical microscopy for 3D ultrastructure.
• Exploration of SPM-based 3D microscopy and its integration with optical methods.
• Comparative analysis of the strengths and weaknesses of diverse 3D microscopy approaches.

Conclusions:

• A range of advanced 3D microscopy techniques are available for detailed ultrastructural analysis in biomedical research.
• Each technique offers unique advantages and disadvantages, requiring careful selection based on research needs.
• Further development and integration of these methods promise enhanced capabilities for biological structure investigation.