English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
 
 
DownloadE-Mail
  On the quantification of intracellular proteins in multifluorescence-labeled rat brain slices using slide-based cytometry

Reinert, A., Mittag, A., Reinert, T., Tárnok, A., Arendt, T., & Morawski, M. (2011). On the quantification of intracellular proteins in multifluorescence-labeled rat brain slices using slide-based cytometry. Cytometry Part A, 79A(6), 485-491. doi:10.1002/cyto.a.21047.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/21.11116/0000-0004-CE07-D Version Permalink: http://hdl.handle.net/21.11116/0000-0004-CE08-C
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Reinert, Anja1, Author
Mittag, Anja1, Author
Reinert, Tilo1, Author              
Tárnok, Attila1, Author
Arendt, Thomas1, Author
Morawski, Markus1, Author
Affiliations:
1External Organizations, ou_persistent22              

Content

show
hide
Free keywords: laser scanning cytometry; slide‐based cytometry; brain; neurons; quantification; perineuronal net; immuno‐/lectinhistochemistry; iron protein ferritin H
 Abstract: In several brain regions, a subpopulation of neurons exists being characterized by the expression of a peculiar form of extracellular matrix, a so‐called perineuronal net (PNN). We have previously shown that the PNN can bind large amounts of iron due to its polyanionic charge. Because free iron can generate reactive oxygen species thus being potentially toxic, the PNN may have a protective function by “scavenging” this free iron. Because of this ability, we have hypothesized that PNN‐related neurons have an altered iron‐specific metabolism. Thus, to compare the intracellular concentrations of iron containing proteins, specifically, the iron storage protein ferritin H between neurons with and without a PNN, we have used slide‐based cytometry with image‐based threshold‐boundary cell detection on brain sections. In tissue sections, the integrity of the extracellular matrix, especially the characteristic PNNs, is preserved, which is necessary for the identification of the two neuronal subpopulations. A multilabeling approach was chosen to select neurons (neuronal marker NeuN), to classify the neurons according to their subtype (matrix marker Wisteria floribunda agglutinin), and to quantify the protein concentration (protein marker). Using this novel method, we were able to detect a relative difference in protein concentration as low as 12% between the two subpopulations of neurons in the neuronal population of the rat parietal cortex. © 2011 International Society for Advancement of Cytometry

Details

show
hide
Language(s): eng - English
 Dates: 2011-01-202010-06-222011-02-162011-03-182011-06
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/cyto.a.21047
BibTex Citekey: Reinert:2011b
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Cytometry Part A
  Other : Cytom. Part A
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Hoboken, N.J. : Wiley-Liss
Pages: - Volume / Issue: 79A (6) Sequence Number: - Start / End Page: 485 - 491 Identifier: ISSN: 1552-4922
CoNE: https://pure.mpg.de/cone/journals/resource/954925582174