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Method Article
Shaina Robbins
Thomas Jefferson University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-7885-743X
Sirisha Achanta
Thomas Jefferson University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-3833-5714
Rajanikanth Vadigepalli
Thomas Jefferson University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-8405-1037
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This protocol demonstrates the use of Laser Capture Microdissection (LCM), a precise cell isolation technique, for acquiring single neurons from right atrial ganglionated plexus (RAGP) porcine heart tissue in sync with image acquisition to enable 3D location tracking of collected samples. Fresh frozen RAGP tissue sectioned and stained for neurons (Nissl stain) is processed through LCM for sample collection. The LCM workflow is comprised of four steps: instrument setup, slide loading and tissue inspection, microdissection and image acquisition and sample processing and storage. LCM collected samples were processed for gene expression experiments : High-throughput-qRT-PCR or Single cell RNA sequencing. Gene expression data along with 3D sample tracking was used to generate a functional map of porcine RAGP. The techniques described in this protocol can be adapted to a wide variety of sample and tissue types with minor modifications. This protocol is a part of a protocol pipeline that includes cryosectioning and staining protocols upstream and Tissue Mapper protocol, High-Throughput q-RT-PCR and RNA Seq protocols downstream.
Keywords
Laser Capture Microdissection, LCM, Single cell, Neuron
Figure 1
Figure 2
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This is a list of supplementary files associated with this preprint. Click to download.
- LaserCaptureMicrodissectionLCMand3DSampleTrackingProtocol.pdf
PDF version of the protocol with figures integrated in the document.
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Posted 08 Jun, 2021
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An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Shaina Robbins
Thomas Jefferson University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-7885-743X
Sirisha Achanta
Thomas Jefferson University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-3833-5714
Rajanikanth Vadigepalli
Thomas Jefferson University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-8405-1037
Version History
CURRENT STATUS: Posted
Version 1
Posted 08 Jun, 2021
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This protocol demonstrates the use of Laser Capture Microdissection (LCM), a precise cell isolation technique, for acquiring single neurons from right atrial ganglionated plexus (RAGP) porcine heart tissue in sync with image acquisition to enable 3D location tracking of collected samples. Fresh frozen RAGP tissue sectioned and stained for neurons (Nissl stain) is processed through LCM for sample collection. The LCM workflow is comprised of four steps: instrument setup, slide loading and tissue inspection, microdissection and image acquisition and sample processing and storage. LCM collected samples were processed for gene expression experiments : High-throughput-qRT-PCR or Single cell RNA sequencing. Gene expression data along with 3D sample tracking was used to generate a functional map of porcine RAGP. The techniques described in this protocol can be adapted to a wide variety of sample and tissue types with minor modifications. This protocol is a part of a protocol pipeline that includes cryosectioning and staining protocols upstream and Tissue Mapper protocol, High-Throughput q-RT-PCR and RNA Seq protocols downstream.
Figure 1
Figure 2
N/A
This is a list of supplementary files associated with this preprint. Click to download.
- LaserCaptureMicrodissectionLCMand3DSampleTrackingProtocol.pdf
PDF version of the protocol with figures integrated in the document.
Loading...