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2017 ; 19
(ä): 7
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3D Printer Generated Tissue iMolds for Cleared Tissue Using Single- and
Multi-Photon Microscopy for Deep Tissue Evaluation
#MMPMID28690429
Miller SJ
; Rothstein JD
Biol Proced Online
2017[]; 19
(ä): 7
PMID28690429
show ga
BACKGROUND: Pathological analyses and methodology has recently undergone a
dramatic revolution. With the creation of tissue clearing methods such as CLARITY
and CUBIC, groups can now achieve complete transparency in tissue samples in
nano-porous hydrogels. Cleared tissue is then imagined in a semi-aqueous medium
that matches the refractive index of the objective being used. However, one major
challenge is the ability to control tissue movement during imaging and to
relocate precise locations post sequential clearing and re-staining. METHODS:
Using 3D printers, we designed tissue molds that fit precisely around the
specimen being imaged. First, images are taken of the specimen, followed by
importing and design of a structural mold, then printed with affordable plastics
by a 3D printer. RESULTS: With our novel design, we have innovated tissue molds
called innovative molds (iMolds) that can be generated in any laboratory and are
customized for any organ, tissue, or bone matter being imaged. Furthermore, the
inexpensive and reusable tissue molds are made compatible for any microscope such
as single and multi-photon confocal with varying stage dimensions. Excitingly,
iMolds can also be generated to hold multiple organs in one mold, making
reconstruction and imaging much easier. CONCLUSIONS: Taken together, with iMolds
it is now possible to image cleared tissue in clearing medium while limiting
movement and being able to relocate precise anatomical and cellular locations on
sequential imaging events in any basic laboratory. This system provides great
potential for screening widespread effects of therapeutics and disease across
entire organ systems.