Analysis of Transcription Factor mRNAs in Identified Oxytocin and Vasopressin Magnocellular Neurons Isolated by Laser Capture Microdissection

Madison Humerick; Jeffrey Hanson; Jaime Rodriguez-Canales; Daniel Lubelski; Omar M. Rashid; Yasmmyn D. Salinas; YiJun Shi; Todd Ponzio; Raymond Fields; Michael R. Emmert-Buck; Harold Gainer

doi:10.1371/journal.pone.0069407

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Peer-reviewed

Research Article

Analysis of Transcription Factor mRNAs in Identified Oxytocin and Vasopressin Magnocellular Neurons Isolated by Laser Capture Microdissection

Madison Humerick ,

Contributed equally to this work with: Madison Humerick, Jeffrey Hanson

Current address: West Virginia University School of Medicine, Morgantown, West Virginia, United States of America

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Jeffrey Hanson ,

Contributed equally to this work with: Madison Humerick, Jeffrey Hanson

Affiliation Laser Microdissection Core, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Jaime Rodriguez-Canales,

Current address: Department of Translational Molecular Pathology, University of Texas - MD Anderson Cancer Center, Houston, Texas, United States of America

Affiliation Laser Microdissection Core, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Daniel Lubelski,

Current address: Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, United States of America

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Omar M. Rashid,

Current address: New York Medical College, Valhalla, New York, United States of America

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Yasmmyn D. Salinas,

Current address: Yale School of Public Health, Yale University, New Haven, Connecticut, United States of America

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
YiJun Shi,

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Todd Ponzio,

Current address: Office of Technology Transfer, Naval Medical Research Center, Silver Spring, Maryland, United States of America

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Raymond Fields,

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Michael R. Emmert-Buck,

Affiliation Laser Microdissection Core, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America
⨯
Harold Gainer

* E-mail: gainerh@ninds.nih.gov

Affiliation Laboratory of Neurochemistry, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland, United States of America
⨯

Analysis of Transcription Factor mRNAs in Identified Oxytocin and Vasopressin Magnocellular Neurons Isolated by Laser Capture Microdissection

Madison Humerick,
Jeffrey Hanson,
Jaime Rodriguez-Canales,
Daniel Lubelski,
Omar M. Rashid,
Yasmmyn D. Salinas,
YiJun Shi,
Todd Ponzio,
Raymond Fields,
Michael R. Emmert-Buck

Published: July 24, 2013
https://doi.org/10.1371/journal.pone.0069407

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RE: Response to query re preparation of slides for LCM

Posted by hgainer on 28 Aug 2013 at 17:42 GMT

My response is:

We do not wash the slides with water, & I don't recall any opaqueness that interfered with the LCM.
You could also consult the first author, M. Humerick via msteven8@mix.wvu.ed

We use the following protocol, the first wash is in 75% ethanol as follows:
1. Using the solutions prepared on the day of use, in a fume hood perform the following steps:
a. 75% Ethanol for 30 sec
b. 95% Ethanol for 5 sec
c. 100% Ethanol for 5 sec
d. 100% Ethanol for 5 sec
e. Xylene for 30 sec
f. Allow Xylene to completely evaporate
i. This usually takes 2-5 min.

The above is written in Table S2 in Supplements.

Good luck,
H. Gainer

No competing interests declared.

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Analysis of Transcription Factor mRNAs in Identified Oxytocin and Vasopressin Magnocellular Neurons Isolated by Laser Capture Microdissection

Analysis of Transcription Factor mRNAs in Identified Oxytocin and Vasopressin Magnocellular Neurons Isolated by Laser Capture Microdissection

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RE: Response to query re preparation of slides for LCM

Posted by hgainer on 28 Aug 2013 at 17:42 GMT