Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy
Summary: Nuclear pore complexes are pathways for nuclear-cytoplasmic communication that participate in chromatin organization. Here, we present a protocol to image and quantify the number of nuclear pore complexes in cells. We describe steps for cell plating and culture, immunofluorescence detection...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723005191 |
| _version_ | 1797729855124537344 |
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| author | Jocelyn D. Mich-Basso Bernhard Kühn |
| author_facet | Jocelyn D. Mich-Basso Bernhard Kühn |
| author_sort | Jocelyn D. Mich-Basso |
| collection | DOAJ |
| description | Summary: Nuclear pore complexes are pathways for nuclear-cytoplasmic communication that participate in chromatin organization. Here, we present a protocol to image and quantify the number of nuclear pore complexes in cells. We describe steps for cell plating and culture, immunofluorescence detection, and confocal microscopy visualization of nuclear pore complexes. We then detail quantification and 3D data analysis. This protocol utilizes digital thresholding under human supervision for quantification of nuclear pore complexes.For complete details on the use and execution of this protocol, please refer to Han et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-12T11:35:42Z |
| format | Article |
| id | doaj.art-a4bcc800f9a044968cdfccf887935b5a |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-12T11:35:42Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-a4bcc800f9a044968cdfccf887935b5a2023-09-01T05:03:09ZengElsevierSTAR Protocols2666-16672023-09-0143102552Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopyJocelyn D. Mich-Basso0Bernhard Kühn1Division of Cardiology, Pediatric Institute for Heart Regeneration and Therapeutics (I-HRT), UPMC Children’s Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA; Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15219, USADivision of Cardiology, Pediatric Institute for Heart Regeneration and Therapeutics (I-HRT), UPMC Children’s Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA; Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15219, USA; Corresponding authorSummary: Nuclear pore complexes are pathways for nuclear-cytoplasmic communication that participate in chromatin organization. Here, we present a protocol to image and quantify the number of nuclear pore complexes in cells. We describe steps for cell plating and culture, immunofluorescence detection, and confocal microscopy visualization of nuclear pore complexes. We then detail quantification and 3D data analysis. This protocol utilizes digital thresholding under human supervision for quantification of nuclear pore complexes.For complete details on the use and execution of this protocol, please refer to Han et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723005191Cell BiologyMicroscopy |
| spellingShingle | Jocelyn D. Mich-Basso Bernhard Kühn Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy STAR Protocols Cell Biology Microscopy |
| title | Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy |
| title_full | Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy |
| title_fullStr | Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy |
| title_full_unstemmed | Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy |
| title_short | Protocol to image and quantify nuclear pore complexes using high-resolution laser scanning confocal microscopy |
| title_sort | protocol to image and quantify nuclear pore complexes using high resolution laser scanning confocal microscopy |
| topic | Cell Biology Microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723005191 |
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