Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.

Automatic operations of multi-functional and time-lapse live-cell imaging are necessary for the biomedical science community to study active, multi-faceted, and long-term biological phenomena. To achieve automatic control, most existing solutions often require the purchase of extra software programs...

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Main Authors: Qin Luo, Justin Zhang, Miao Huang, Gaoming Lin, Mai Tanaka, Sharon Lepler, Juan Guan, Dietmar Siemann, Xin Tang
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2022-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0266098
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author Qin Luo
Justin Zhang
Miao Huang
Gaoming Lin
Mai Tanaka
Sharon Lepler
Juan Guan
Dietmar Siemann
Xin Tang
author_facet Qin Luo
Justin Zhang
Miao Huang
Gaoming Lin
Mai Tanaka
Sharon Lepler
Juan Guan
Dietmar Siemann
Xin Tang
author_sort Qin Luo
collection DOAJ
description Automatic operations of multi-functional and time-lapse live-cell imaging are necessary for the biomedical science community to study active, multi-faceted, and long-term biological phenomena. To achieve automatic control, most existing solutions often require the purchase of extra software programs and hardware that rely on the manufacturers' own specifications. However, these software programs are usually non-user-programmable and unaffordable for many laboratories. To address this unmet need, we have developed a novel open-source software program, titled Automatic Multi-functional Integration Program (AMFIP), as a new Java-based and hardware-independent system that provides proven advantages over existing alternatives to the scientific community. Without extra hardware, AMFIP enables the functional synchronization of the μManager software platform, the Nikon NIS-Elements platform, and other 3rd party software to achieve automatic operations of most commercially available microscopy systems, including but not limited to those from Nikon. AMFIP provides a user-friendly and programmable graphical user interface (GUI), opening the door to expanding the customizability for myriad hardware and software systems according to user-specific experimental requirements and environments. To validate the intended purposes of developing AMFIP, we applied it to elucidate the question whether single cells, prior to their full spreading, can sense and respond to a soft solid substrate, and if so, how does the interaction depend on the cell spreading time and the stiffness of the substrate. Using a CRISPR/Cas9-engineered human epithelial Beas2B (B2B) cell line that expresses mNeonGreen2-tagged mechanosensitive Yes-associated protein (YAP), we show that single B2B cells develop distinct substrate-stiffness-dependent YAP expressions within 10 hours at most on the substrate, suggesting that cells are able to sense, distinguish, and respond to mechanical cues prior to the establishment of full cell spreading. In summary, AMFIP provides a reliable, open-source, and cost-free solution that has the validated long-term utility to satisfy the need of automatic imaging operations in the scientific community.
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spelling doaj.art-d55e0c06eea34a4b8a382a21fddf6b872022-12-22T02:52:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-01177e026609810.1371/journal.pone.0266098Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.Qin LuoJustin ZhangMiao HuangGaoming LinMai TanakaSharon LeplerJuan GuanDietmar SiemannXin TangAutomatic operations of multi-functional and time-lapse live-cell imaging are necessary for the biomedical science community to study active, multi-faceted, and long-term biological phenomena. To achieve automatic control, most existing solutions often require the purchase of extra software programs and hardware that rely on the manufacturers' own specifications. However, these software programs are usually non-user-programmable and unaffordable for many laboratories. To address this unmet need, we have developed a novel open-source software program, titled Automatic Multi-functional Integration Program (AMFIP), as a new Java-based and hardware-independent system that provides proven advantages over existing alternatives to the scientific community. Without extra hardware, AMFIP enables the functional synchronization of the μManager software platform, the Nikon NIS-Elements platform, and other 3rd party software to achieve automatic operations of most commercially available microscopy systems, including but not limited to those from Nikon. AMFIP provides a user-friendly and programmable graphical user interface (GUI), opening the door to expanding the customizability for myriad hardware and software systems according to user-specific experimental requirements and environments. To validate the intended purposes of developing AMFIP, we applied it to elucidate the question whether single cells, prior to their full spreading, can sense and respond to a soft solid substrate, and if so, how does the interaction depend on the cell spreading time and the stiffness of the substrate. Using a CRISPR/Cas9-engineered human epithelial Beas2B (B2B) cell line that expresses mNeonGreen2-tagged mechanosensitive Yes-associated protein (YAP), we show that single B2B cells develop distinct substrate-stiffness-dependent YAP expressions within 10 hours at most on the substrate, suggesting that cells are able to sense, distinguish, and respond to mechanical cues prior to the establishment of full cell spreading. In summary, AMFIP provides a reliable, open-source, and cost-free solution that has the validated long-term utility to satisfy the need of automatic imaging operations in the scientific community.https://doi.org/10.1371/journal.pone.0266098
spellingShingle Qin Luo
Justin Zhang
Miao Huang
Gaoming Lin
Mai Tanaka
Sharon Lepler
Juan Guan
Dietmar Siemann
Xin Tang
Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.
PLoS ONE
title Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.
title_full Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.
title_fullStr Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.
title_full_unstemmed Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.
title_short Automatic Multi-functional Integration Program (AMFIP) towards all-optical mechano-electrophysiology interrogation.
title_sort automatic multi functional integration program amfip towards all optical mechano electrophysiology interrogation
url https://doi.org/10.1371/journal.pone.0266098
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