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Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes
Local manipulation of complex tissues at the single-cell level is challenging and requires excellent sealing between the specimen and the micromanipulation device. Here, biological applications for a recently developed loading technique for a force- and pressure-controlled fluidic force microscope m...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5897369/ https://www.ncbi.nlm.nih.gov/pubmed/29651136 http://dx.doi.org/10.1038/s41598-018-24255-9 |
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author | Roder, Phillip Hille, Carsten |
author_facet | Roder, Phillip Hille, Carsten |
author_sort | Roder, Phillip |
collection | PubMed |
description | Local manipulation of complex tissues at the single-cell level is challenging and requires excellent sealing between the specimen and the micromanipulation device. Here, biological applications for a recently developed loading technique for a force- and pressure-controlled fluidic force microscope micropipette are described. This technique allows for the exact positioning and precise spatiotemporal control of liquid delivery. The feasibility of a local loading technique for tissue applications was investigated using two fluorescent dyes, with which local loading behaviour could be optically visualised. Thus, homogeneous intracellular distribution of CellTracker Red and accumulation of SYTO 9 Green within nuclei was realised in single cells of a tissue preparation. Subsequently, physiological micromanipulation experiments were performed. Salivary gland tissue was pre-incubated with the Ca(2+)-sensitive dye OGB-1. An intracellular Ca(2+) rise was then initiated at the single-cell level by applying dopamine via micropipette. When pre-incubating tissue with the nitric oxide (NO)-sensitive dye DAF-FM, NO release and intercellular NO diffusion was observed after local application of the NO donor SNP. Finally, local micromanipulation of a well-defined area along irregularly shaped cell surfaces of complex biosystems was shown for the first time for the fluidic force microscope micropipette. Thus, this technique is a promising tool for the investigation of the spatiotemporal effects of locally applied substances in complex tissues. |
format | Online Article Text |
id | pubmed-5897369 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-58973692018-04-20 Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes Roder, Phillip Hille, Carsten Sci Rep Article Local manipulation of complex tissues at the single-cell level is challenging and requires excellent sealing between the specimen and the micromanipulation device. Here, biological applications for a recently developed loading technique for a force- and pressure-controlled fluidic force microscope micropipette are described. This technique allows for the exact positioning and precise spatiotemporal control of liquid delivery. The feasibility of a local loading technique for tissue applications was investigated using two fluorescent dyes, with which local loading behaviour could be optically visualised. Thus, homogeneous intracellular distribution of CellTracker Red and accumulation of SYTO 9 Green within nuclei was realised in single cells of a tissue preparation. Subsequently, physiological micromanipulation experiments were performed. Salivary gland tissue was pre-incubated with the Ca(2+)-sensitive dye OGB-1. An intracellular Ca(2+) rise was then initiated at the single-cell level by applying dopamine via micropipette. When pre-incubating tissue with the nitric oxide (NO)-sensitive dye DAF-FM, NO release and intercellular NO diffusion was observed after local application of the NO donor SNP. Finally, local micromanipulation of a well-defined area along irregularly shaped cell surfaces of complex biosystems was shown for the first time for the fluidic force microscope micropipette. Thus, this technique is a promising tool for the investigation of the spatiotemporal effects of locally applied substances in complex tissues. Nature Publishing Group UK 2018-04-12 /pmc/articles/PMC5897369/ /pubmed/29651136 http://dx.doi.org/10.1038/s41598-018-24255-9 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Roder, Phillip Hille, Carsten Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes |
title | Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes |
title_full | Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes |
title_fullStr | Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes |
title_full_unstemmed | Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes |
title_short | Local tissue manipulation via a force- and pressure-controlled AFM micropipette for analysis of cellular processes |
title_sort | local tissue manipulation via a force- and pressure-controlled afm micropipette for analysis of cellular processes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5897369/ https://www.ncbi.nlm.nih.gov/pubmed/29651136 http://dx.doi.org/10.1038/s41598-018-24255-9 |
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