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Navigating across multi-dimensional space of tissue clearing parameters.

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Optical tissue clearing renders thick biological samples transparent for advanced light microscopy. Choosing the optimal clearing method requires careful consideration of sample type, imaging needs, and available resources.

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Area of Science:

  • Biomedical Engineering
  • Microscopy Techniques
  • Histology

Background:

  • Optical tissue clearing, first described in 1914, gained prominence in the 21st century due to advancements in imaging instrumentation.
  • Modern microscopy, computing, and software enable the imaging of centimeter-thick samples, driving the development of diverse tissue clearing protocols.
  • Existing protocols utilize organic solvents, water-miscible agents, electric fields, or perfusion, with no single method being universally optimal.

Purpose of the Study:

  • To provide a practical overview of various optical tissue clearing techniques.
  • To guide non-experts in selecting the most suitable tissue clearing method for their specific research needs.
  • To highlight key parameters influencing the choice of a tissue clearing protocol.

Main Methods:

  • Review of diverse physico-chemical treatments for optical tissue clearing.
  • Analysis of protocols involving organic solvents, detergents, chaotropic agents, electric fields, and perfusion.
  • Consideration of factors such as image quality, sample characteristics, labeling, instrumentation, budget, and time.

Main Results:

  • Numerous tissue clearing protocols exist, varying in chemical agents and procedural requirements.
  • The selection of a tissue clearing method is contingent upon multiple interdependent factors.
  • Inappropriate protocol selection can lead to significant time expenditure.

Conclusions:

  • No single optical tissue clearing method is universally superior; method selection is context-dependent.
  • Careful evaluation of parameters like image resolution, sample size, label compatibility, and available resources is crucial for efficient tissue clearing.
  • This review aims to facilitate informed decision-making for researchers utilizing optical tissue clearing techniques.