Microdissection of gonadal tissues for gene expression analyses

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Standard

Microdissection of gonadal tissues for gene expression analyses. / Jørgensen, Anne; Dalgaard, Marlene Danner; Sonne, Si Brask.

Laser capture microdissection: Methods and protocols. ed. / Graeme I. Murray. Vol. 755 Humana Press, 2011. p. 307-13 (Methods in Molecular Biology).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Harvard

Jørgensen, A, Dalgaard, MD & Sonne, SB 2011, Microdissection of gonadal tissues for gene expression analyses. in GI Murray (ed.), Laser capture microdissection: Methods and protocols. vol. 755, Humana Press, Methods in Molecular Biology, pp. 307-13. https://doi.org/10.1007/978-1-61779-163-5_26

APA

Jørgensen, A., Dalgaard, M. D., & Sonne, S. B. (2011). Microdissection of gonadal tissues for gene expression analyses. In G. I. Murray (Ed.), Laser capture microdissection: Methods and protocols (Vol. 755, pp. 307-13). Humana Press. Methods in Molecular Biology https://doi.org/10.1007/978-1-61779-163-5_26

Vancouver

Jørgensen A, Dalgaard MD, Sonne SB. Microdissection of gonadal tissues for gene expression analyses. In Murray GI, editor, Laser capture microdissection: Methods and protocols. Vol. 755. Humana Press. 2011. p. 307-13. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-163-5_26

Author

Jørgensen, Anne ; Dalgaard, Marlene Danner ; Sonne, Si Brask. / Microdissection of gonadal tissues for gene expression analyses. Laser capture microdissection: Methods and protocols. editor / Graeme I. Murray. Vol. 755 Humana Press, 2011. pp. 307-13 (Methods in Molecular Biology).

Bibtex

@inbook{932f7f4bb55349b7858a08cd0046bc81,
title = "Microdissection of gonadal tissues for gene expression analyses",
abstract = "Laser microdissection permits isolation of specific cell types from tissue sections or cell cultures. This may be beneficial when investigating the role of specific cells in a complex tissue or organ. In tissues with easily distinguishable morphology, a simple hematoxylin staining is sufficient, but in most cases a more specific staining is required to identify which cells to microdissect. We have established two staining protocols for frozen sections (1) Oil red O, which stains lipid droplet in fat cells and steroid-producing cells and (2) NBT BCIP, which stains cells expressing an alkaline phosphatase enzyme, such as fetal germ cells, testicular carcinoma in situ cells, and putatively also other early stem cell populations. We have applied these protocols for microdissection of rat Leydig cells, fetal human and zebrafish germ cells, and human testicular germ cell tumors, but the staining protocols could also be used in other species and for other cell types containing lipid droplets or expressing alkaline phosphatase. Both protocols ensure a morphology that enables microdissection of single cells with RNA quality sufficient for subsequent gene expression analysis. However, RNA yields after microdissection and purification are small, and therefore, two rounds of linear amplification are recommended prior to gene expression analysis.",
keywords = "Animals, Azo Compounds, Coloring Agents, Gene Expression Profiling, Humans, Lasers, Male, Microdissection, Oligonucleotide Array Sequence Analysis, RNA, Rats, Reverse Transcriptase Polymerase Chain Reaction, Staining and Labeling, Testis, Tissue Fixation, Alkaline phosphate, Oil red O, Lipid droplets, NBT BCIP, histological staining, fetal gonads, Gene Expression",
author = "Anne J{\o}rgensen and Dalgaard, {Marlene Danner} and Sonne, {Si Brask}",
year = "2011",
doi = "10.1007/978-1-61779-163-5_26",
language = "English",
isbn = "978-1-61779-162-8",
volume = "755",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "307--13",
editor = "Murray, {Graeme I.}",
booktitle = "Laser capture microdissection",
address = "United States",

}

RIS

TY - CHAP

T1 - Microdissection of gonadal tissues for gene expression analyses

AU - Jørgensen, Anne

AU - Dalgaard, Marlene Danner

AU - Sonne, Si Brask

PY - 2011

Y1 - 2011

N2 - Laser microdissection permits isolation of specific cell types from tissue sections or cell cultures. This may be beneficial when investigating the role of specific cells in a complex tissue or organ. In tissues with easily distinguishable morphology, a simple hematoxylin staining is sufficient, but in most cases a more specific staining is required to identify which cells to microdissect. We have established two staining protocols for frozen sections (1) Oil red O, which stains lipid droplet in fat cells and steroid-producing cells and (2) NBT BCIP, which stains cells expressing an alkaline phosphatase enzyme, such as fetal germ cells, testicular carcinoma in situ cells, and putatively also other early stem cell populations. We have applied these protocols for microdissection of rat Leydig cells, fetal human and zebrafish germ cells, and human testicular germ cell tumors, but the staining protocols could also be used in other species and for other cell types containing lipid droplets or expressing alkaline phosphatase. Both protocols ensure a morphology that enables microdissection of single cells with RNA quality sufficient for subsequent gene expression analysis. However, RNA yields after microdissection and purification are small, and therefore, two rounds of linear amplification are recommended prior to gene expression analysis.

AB - Laser microdissection permits isolation of specific cell types from tissue sections or cell cultures. This may be beneficial when investigating the role of specific cells in a complex tissue or organ. In tissues with easily distinguishable morphology, a simple hematoxylin staining is sufficient, but in most cases a more specific staining is required to identify which cells to microdissect. We have established two staining protocols for frozen sections (1) Oil red O, which stains lipid droplet in fat cells and steroid-producing cells and (2) NBT BCIP, which stains cells expressing an alkaline phosphatase enzyme, such as fetal germ cells, testicular carcinoma in situ cells, and putatively also other early stem cell populations. We have applied these protocols for microdissection of rat Leydig cells, fetal human and zebrafish germ cells, and human testicular germ cell tumors, but the staining protocols could also be used in other species and for other cell types containing lipid droplets or expressing alkaline phosphatase. Both protocols ensure a morphology that enables microdissection of single cells with RNA quality sufficient for subsequent gene expression analysis. However, RNA yields after microdissection and purification are small, and therefore, two rounds of linear amplification are recommended prior to gene expression analysis.

KW - Animals

KW - Azo Compounds

KW - Coloring Agents

KW - Gene Expression Profiling

KW - Humans

KW - Lasers

KW - Male

KW - Microdissection

KW - Oligonucleotide Array Sequence Analysis

KW - RNA

KW - Rats

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Staining and Labeling

KW - Testis

KW - Tissue Fixation

KW - Alkaline phosphate

KW - Oil red O

KW - Lipid droplets

KW - NBT BCIP

KW - histological staining

KW - fetal gonads

KW - Gene Expression

U2 - 10.1007/978-1-61779-163-5_26

DO - 10.1007/978-1-61779-163-5_26

M3 - Book chapter

C2 - 21761315

SN - 978-1-61779-162-8

VL - 755

T3 - Methods in Molecular Biology

SP - 307

EP - 313

BT - Laser capture microdissection

A2 - Murray, Graeme I.

PB - Humana Press

ER -

ID: 41989447