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3D Printer Generated Tissue iMolds for Cleared Tissue Using Single- and Multi-Photon Microscopy for Deep Tissue

Sean J Miller^1,2,3, Jeffrey D Rothstein^1,2,3

¹Department of Neurology, Johns Hopkins University School of Medicine, Johns Hopkins Univ., 855 N Wolfe Street, Room 250.18, Baltimore, MD 21205 USA.

Biological Procedures Online

|July 11, 2017

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This summary is machine-generated.

Innovative molds (iMolds) enable stable imaging of cleared tissues, overcoming challenges in precise relocation for advanced pathological analysis. These 3D-printed, customizable molds ensure consistent results in any lab.

Area of Science:

Pathology
Biotechnology
Microscopy

Background:

Tissue clearing techniques like CLARITY and CUBIC enable transparent tissue samples.
A key challenge is stabilizing cleared tissue during imaging and precise relocation for sequential analysis.
Current methods lack robust solutions for tissue movement control and accurate spatial referencing.

Purpose of the Study:

To develop a novel solution for stabilizing cleared tissue during imaging.
To enable precise relocation of anatomical and cellular locations in cleared tissues.
To facilitate advanced pathological analysis and screening of organ-wide effects.

Main Methods:

3D printing technology was utilized to design and fabricate custom tissue molds (iMolds).
iMolds are precisely fitted around specimens, allowing for initial imaging before mold creation.

Keywords:

3D printer CLARITY Cleared tissue Medicine Microscopy Tissue molds

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The molds are printed using affordable plastics and are reusable.

Main Results:

Innovative molds (iMolds) were successfully developed for customizable tissue stabilization.
iMolds are compatible with various microscopes (e.g., confocal) and stage dimensions.
The system allows for imaging of multiple organs within a single mold, simplifying reconstruction.

Conclusions:

iMolds provide a stable platform for imaging cleared tissues, minimizing movement artifacts.
Precise anatomical and cellular locations can be reliably relocated across sequential imaging events.
This technology supports widespread screening of therapeutic and disease effects across organ systems in basic laboratories.