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An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization

Recent development in 3D printing technology has opened an exciting possibility for manufacturing 3D devices on one’s desktop. We used 3D modeling programs to design 3D models of a tissue-handling system and these models were “printed” in a stereolithography (SLA) 3D printer to create precision hist...

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Detalles Bibliográficos
Autores principales: Anthony, Giovanni, Lee, Ju-Ahng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4973989/
https://www.ncbi.nlm.nih.gov/pubmed/27489962
http://dx.doi.org/10.1371/journal.pone.0159991
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author Anthony, Giovanni
Lee, Ju-Ahng
author_facet Anthony, Giovanni
Lee, Ju-Ahng
author_sort Anthony, Giovanni
collection PubMed
description Recent development in 3D printing technology has opened an exciting possibility for manufacturing 3D devices on one’s desktop. We used 3D modeling programs to design 3D models of a tissue-handling system and these models were “printed” in a stereolithography (SLA) 3D printer to create precision histology devices that are particularly useful to handle multiple samples with small dimensions in parallel. Our system has been successfully tested for in situ hybridization of zebrafish embryos. Some of the notable features include: (1) A conveniently transferrable chamber with 6 mesh-bottomed wells, each of which can hold dozens of zebrafish embryos. This design allows up to 6 different samples to be treated per chamber. (2) Each chamber sits in a well of a standard 6-well tissue culture plate. Thus, up to 36 different samples can be processed in tandem using a single 6 well plate. (3) Precisely fitting lids prevent solution evaporation and condensation, even at high temperatures for an extended period of time: i.e., overnight riboprobe hybridization. (4) Flat bottom mesh maximizes the consistent treatment of individual tissue samples. (5) A magnet-based lifter was created to handle up to 6 chambers (= 36 samples) in unison. (6) The largely transparent resin aids in convenient visual inspection both with eyes and using a stereomicroscope. (7) Surface engraved labeling enables an accurate tracking of different samples. (8) The dimension of wells and chambers minimizes the required amount of precious reagents. (9) Flexible parametric modeling enables an easy redesign of the 3D models to handle larger or more numerous samples. Precise dimensions of 3D models and demonstration of how we use our devices in whole mount in situ hybridization are presented. We also provide detailed information on the modeling software, 3D printing tips, as well as 3D files that can be used with any 3D printer.
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spelling pubmed-49739892016-08-18 An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization Anthony, Giovanni Lee, Ju-Ahng PLoS One Research Article Recent development in 3D printing technology has opened an exciting possibility for manufacturing 3D devices on one’s desktop. We used 3D modeling programs to design 3D models of a tissue-handling system and these models were “printed” in a stereolithography (SLA) 3D printer to create precision histology devices that are particularly useful to handle multiple samples with small dimensions in parallel. Our system has been successfully tested for in situ hybridization of zebrafish embryos. Some of the notable features include: (1) A conveniently transferrable chamber with 6 mesh-bottomed wells, each of which can hold dozens of zebrafish embryos. This design allows up to 6 different samples to be treated per chamber. (2) Each chamber sits in a well of a standard 6-well tissue culture plate. Thus, up to 36 different samples can be processed in tandem using a single 6 well plate. (3) Precisely fitting lids prevent solution evaporation and condensation, even at high temperatures for an extended period of time: i.e., overnight riboprobe hybridization. (4) Flat bottom mesh maximizes the consistent treatment of individual tissue samples. (5) A magnet-based lifter was created to handle up to 6 chambers (= 36 samples) in unison. (6) The largely transparent resin aids in convenient visual inspection both with eyes and using a stereomicroscope. (7) Surface engraved labeling enables an accurate tracking of different samples. (8) The dimension of wells and chambers minimizes the required amount of precious reagents. (9) Flexible parametric modeling enables an easy redesign of the 3D models to handle larger or more numerous samples. Precise dimensions of 3D models and demonstration of how we use our devices in whole mount in situ hybridization are presented. We also provide detailed information on the modeling software, 3D printing tips, as well as 3D files that can be used with any 3D printer. Public Library of Science 2016-08-04 /pmc/articles/PMC4973989/ /pubmed/27489962 http://dx.doi.org/10.1371/journal.pone.0159991 Text en © 2016 Anthony, Lee http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Anthony, Giovanni
Lee, Ju-Ahng
An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization
title An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization
title_full An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization
title_fullStr An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization
title_full_unstemmed An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization
title_short An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization
title_sort optimized small tissue handling system for immunohistochemistry and in situ hybridization
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4973989/
https://www.ncbi.nlm.nih.gov/pubmed/27489962
http://dx.doi.org/10.1371/journal.pone.0159991
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