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Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice

BACKGROUND: The new method described here is highly efficient in transferring microinjected mouse eggs (MEs) through the bursa membrane of a surrogate mother mouse to the ampulla of the oviduct without damaging the blood vessels on the bursa membrane. RESULTS: This method causes no loss of blood, an...

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Autores principales: Wen, Guang, Di, Jing, Li, Qian, Chen, Jianling, Jin, Ling, Wang, Cheng, Xu, Sanqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5138171/
https://www.ncbi.nlm.nih.gov/pubmed/28018784
http://dx.doi.org/10.1186/s40064-016-3760-6
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author Wen, Guang
Di, Jing
Li, Qian
Chen, Jianling
Jin, Ling
Wang, Cheng
Xu, Sanqing
author_facet Wen, Guang
Di, Jing
Li, Qian
Chen, Jianling
Jin, Ling
Wang, Cheng
Xu, Sanqing
author_sort Wen, Guang
collection PubMed
description BACKGROUND: The new method described here is highly efficient in transferring microinjected mouse eggs (MEs) through the bursa membrane of a surrogate mother mouse to the ampulla of the oviduct without damaging the blood vessels on the bursa membrane. RESULTS: This method causes no loss of blood, and it produces newborn pups/founders from approximately 70% of the transferred MEs, because only a small hole is made on the blood vessel–free area of the bursa membrane and ampulla of the surrogate mother mouse. The infundibulum remains intact. The small hole on the bursa membrane/ampulla may already heal up before the delivery of the new pups. The method described here consists of a simple operation with a home-assembled drill head holding a self-closing fine forceps on one end, while the drill head assembly body is hooked up with the light housing clamp of a dissecting light microscope. The drill head assembly body can be alternatively hooked/tied up to an appropriate size of clamp (purchased from Home Depot) screwed to any light stand with folding segments. CONCLUSION: This system is able to steadily hold the self-closing fine forceps without shaking and to let the operator use their two hands to steadily hold and quickly insert the pipet carrying the MEs into the ampulla without any delay. Generally MEs stay alive for approximately 15 min at room temperature. The shorter the insertion time is, the more MEs that will survive. Thus, this method may produces more pups/founders. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-016-3760-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-51381712016-12-23 Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice Wen, Guang Di, Jing Li, Qian Chen, Jianling Jin, Ling Wang, Cheng Xu, Sanqing Springerplus Methodology BACKGROUND: The new method described here is highly efficient in transferring microinjected mouse eggs (MEs) through the bursa membrane of a surrogate mother mouse to the ampulla of the oviduct without damaging the blood vessels on the bursa membrane. RESULTS: This method causes no loss of blood, and it produces newborn pups/founders from approximately 70% of the transferred MEs, because only a small hole is made on the blood vessel–free area of the bursa membrane and ampulla of the surrogate mother mouse. The infundibulum remains intact. The small hole on the bursa membrane/ampulla may already heal up before the delivery of the new pups. The method described here consists of a simple operation with a home-assembled drill head holding a self-closing fine forceps on one end, while the drill head assembly body is hooked up with the light housing clamp of a dissecting light microscope. The drill head assembly body can be alternatively hooked/tied up to an appropriate size of clamp (purchased from Home Depot) screwed to any light stand with folding segments. CONCLUSION: This system is able to steadily hold the self-closing fine forceps without shaking and to let the operator use their two hands to steadily hold and quickly insert the pipet carrying the MEs into the ampulla without any delay. Generally MEs stay alive for approximately 15 min at room temperature. The shorter the insertion time is, the more MEs that will survive. Thus, this method may produces more pups/founders. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-016-3760-6) contains supplementary material, which is available to authorized users. Springer International Publishing 2016-12-05 /pmc/articles/PMC5138171/ /pubmed/28018784 http://dx.doi.org/10.1186/s40064-016-3760-6 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Methodology
Wen, Guang
Di, Jing
Li, Qian
Chen, Jianling
Jin, Ling
Wang, Cheng
Xu, Sanqing
Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
title Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
title_full Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
title_fullStr Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
title_full_unstemmed Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
title_short Efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
title_sort efficient method for transfer of microinjected eggs to mouse ampulla for generating transgenic mice
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5138171/
https://www.ncbi.nlm.nih.gov/pubmed/28018784
http://dx.doi.org/10.1186/s40064-016-3760-6
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