Informacja

Drogi użytkowniku, aplikacja do prawidłowego działania wymaga obsługi JavaScript. Proszę włącz obsługę JavaScript w Twojej przeglądarce.

Tytuł pozycji:

High-Temporal-Resolution smFISH Method for Gene Expression Studies in Caenorhabditis elegans Embryos.

Tytuł:
High-Temporal-Resolution smFISH Method for Gene Expression Studies in Caenorhabditis elegans Embryos.
Autorzy:
Charles S; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 313 Ferst Drive NW, Atlanta, Georgia 30332, United States.; Interdisciplinary Program in Bioengineering, Georgia Institute of Technology, 311 Ferst Drive NW, Atlanta, Georgia 30332, United States.
Aubry G; School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Drive NW, Atlanta, Georgia 30332, United States.
Chou HT; School of Biological Sciences, Georgia Institute of Technology, 310 Ferst Drive NW, Atlanta, Georgia 30332, United States.
Paaby AB; School of Biological Sciences, Georgia Institute of Technology, 310 Ferst Drive NW, Atlanta, Georgia 30332, United States.
Lu H; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 313 Ferst Drive NW, Atlanta, Georgia 30332, United States.; Interdisciplinary Program in Bioengineering, Georgia Institute of Technology, 311 Ferst Drive NW, Atlanta, Georgia 30332, United States.; School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Drive NW, Atlanta, Georgia 30332, United States.
Źródło:
Analytical chemistry [Anal Chem] 2021 Jan 26; Vol. 93 (3), pp. 1369-1376. Date of Electronic Publication: 2020 Dec 23.
Typ publikacji:
Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.
Język:
English
Imprint Name(s):
Original Publication: Washington, American Chemical Society.
MeSH Terms:
In Situ Hybridization, Fluorescence*
Caenorhabditis elegans/*genetics
Embryonic Development/*genetics
Animals ; Caenorhabditis elegans/embryology ; Embryo, Nonmammalian
References:
Lab Chip. 2017 Oct 25;17(21):3634-3642. (PMID: 28952622)
Nat Protoc. 2013 Apr;8(4):721-36. (PMID: 23493069)
Proc Natl Acad Sci U S A. 2014 Jul 22;111(29):E2967-76. (PMID: 25002478)
Science. 2015 Apr 24;348(6233):aaa6090. (PMID: 25858977)
Elife. 2018 Sep 27;7:. (PMID: 30260314)
Sci Rep. 2015 May 07;5:10192. (PMID: 25950235)
Curr Top Dev Biol. 2017;123:365-397. (PMID: 28236972)
Anal Chem. 2018 Oct 2;90(19):11470-11477. (PMID: 30125088)
Dis Model Mech. 2017 Jul 1;10(7):923-929. (PMID: 28495673)
Anal Chem. 2011 Sep 15;83(18):7044-52. (PMID: 21809821)
Development. 2019 Mar 25;146(6):. (PMID: 30814118)
Nat Biotechnol. 2018 Nov 12;:. (PMID: 30418432)
Cell. 1988 Feb 12;52(3):311-20. (PMID: 3345562)
Nat Commun. 2019 Aug 2;10(1):3486. (PMID: 31375669)
PLoS One. 2013 Sep 16;8(9):e75120. (PMID: 24066168)
Anal Chem. 1998 Dec 1;70(23):4974-84. (PMID: 21644679)
Lab Chip. 2014 Dec 7;14(23):4513-4522. (PMID: 25257026)
Science. 2019 Sep 20;365(6459):. (PMID: 31488706)
Adv Sci (Weinh). 2018 Mar 08;5(5):1700751. (PMID: 29876206)
Genome Res. 2016 Oct;26(10):1441-1450. (PMID: 27531719)
Cell. 1995 May 19;81(4):611-20. (PMID: 7758115)
Dev Cell. 2012 May 15;22(5):1101-8. (PMID: 22560298)
Nat Methods. 2008 Oct;5(10):877-9. (PMID: 18806792)
Nat Methods. 2013 Nov;10(11):1122-6. (PMID: 24097271)
Nat Methods. 2019 Jun;16(6):533-544. (PMID: 31110282)
Nat Methods. 2020 Jul;17(7):689-693. (PMID: 32541852)
Sci Rep. 2019 Oct 4;9(1):14340. (PMID: 31586133)
Nat Methods. 2013 Apr;10(4):277-8. (PMID: 23538861)
Proc Natl Acad Sci U S A. 2020 Jan 21;117(3):1552-1558. (PMID: 31900360)
Biomicrofluidics. 2019 Nov 01;13(6):064101. (PMID: 31700560)
Nature. 2010 Feb 18;463(7283):913-8. (PMID: 20164922)
Nature. 2019 Apr;568(7751):235-239. (PMID: 30911168)
WormBook. 2012 Dec 13;:1-16. (PMID: 23242966)
Nat Methods. 2011 Feb;8(2):171-176. (PMID: 21186361)
Lab Chip. 2017 Nov 7;17(22):3736-3759. (PMID: 28840220)
Grant Information:
R01 NS115484 United States NS NINDS NIH HHS; R01 NS096581 United States NS NINDS NIH HHS; R21 NS117066 United States NS NINDS NIH HHS; R01 AG056436 United States AG NIA NIH HHS; R01 GM088333 United States GM NIGMS NIH HHS; R35 GM119744 United States GM NIGMS NIH HHS
Entry Date(s):
Date Created: 20201223 Date Completed: 20210218 Latest Revision: 20231103
Update Code:
20240105
PubMed Central ID:
PMC10619480
DOI:
10.1021/acs.analchem.0c02966
PMID:
33355449
Czasopismo naukowe
Recent development in fluorescence-based molecular tools has contributed significantly to developmental studies, including embryogenesis. Many of these tools rely on multiple steps of sample manipulation, so obtaining large sample sizes presents a major challenge as it can be labor-intensive and time-consuming. However, large sample sizes are required to uncover critical aspects of embryogenesis, for example, subtle phenotypic differences or gene expression dynamics. This problem is particularly relevant for single-molecule fluorescence in situ hybridization (smFISH) studies in Caenorhabditis elegans embryogenesis. Microfluidics can help address this issue by allowing a large number of samples and parallelization of experiments. However, performing efficient reagent exchange on chip for large numbers of embryos remains a bottleneck. Here, we present a microfluidic pipeline for large-scale smFISH imaging of C. elegans embryos with minimized labor. We designed embryo traps and engineered a protocol allowing for efficient chemical exchange for hundreds of C. elegans embryos simultaneously. Furthermore, the device design and small footprint optimize imaging throughput by facilitating spatial registration and enabling minimal user input. We conducted the smFISH protocol on chip and demonstrated that image quality is preserved. With one device replacing the equivalent of 10 glass slides of embryos mounted manually, our microfluidic approach greatly increases throughput. Finally, to highlight the capability of our platform to perform longitudinal studies with high temporal resolution, we conducted a temporal analysis of par-1 gene expression in early C. elegans embryos. The method demonstrated here paves the way for systematic high-temporal-resolution studies that will benefit large-scale RNAi and drug screens and in systems beyond C. elegans embryos.

Ta witryna wykorzystuje pliki cookies do przechowywania informacji na Twoim komputerze. Pliki cookies stosujemy w celu świadczenia usług na najwyższym poziomie, w tym w sposób dostosowany do indywidualnych potrzeb. Korzystanie z witryny bez zmiany ustawień dotyczących cookies oznacza, że będą one zamieszczane w Twoim komputerze. W każdym momencie możesz dokonać zmiany ustawień dotyczących cookies