Vamping

Stereology-based automated quantification of fluorescent puncta size and density

Dani Dumitriu, Seth I. Berger, Carine Hamo, Yuko Hara, Megan Bailey, Amarelle Hamo, Yael S. Grossman, William G. Janssen, John Morrison

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The size of dendritic spines and postsynaptic densities (PSDs) is well known to be correlated with molecular and functional characteristics of the synapse. Thus, the development of microscopy methods that allow high throughput quantification and measurement of PSDs is a contemporary need in the field of neurobiology. While the gold standard for measurement of sub-micrometer structures remains electron microscopy (EM), this method is exceedingly laborious and therefore not always feasible. Immunohistochemistry (IHC) is a much faster technique for identifying biological structures such as PSDs, but the fluorescent images resulting from it have traditionally been harder to interpret and quantify. Here, we report on two new image analysis tools that result in accurate size and density measurements of fluorescent puncta. Anti-PSD-95 staining was used to target synapses. The new technique of vamping, using Volume Assisted Measurement of Puncta in 2 and 3 Dimensions (VAMP2D and VAMP3D) respectively, is based on stereological principles. The fully automated image analysis tool was tested on the same subjects for whom we had previously obtained EM measurements of PSD size and/or density. Based on highly consistent results between data obtained by each of these methods, vamping offers an expedient alternative to EM that can nonetheless deliver a high level of accuracy in measuring sub-cellular structures.

Original languageEnglish (US)
Pages (from-to)97-105
Number of pages9
JournalJournal of Neuroscience Methods
Volume209
Issue number1
DOIs
StatePublished - Jul 30 2012
Externally publishedYes

Fingerprint

Post-Synaptic Density
Electron Microscopy
Synapses
Dendritic Spines
Neurobiology
Cellular Structures
Microscopy
Immunohistochemistry
Staining and Labeling

Keywords

  • Confocal microscopy
  • Electron microscopy
  • Image analysis
  • Postsynaptic densities
  • PSD-95
  • Stereology

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Vamping : Stereology-based automated quantification of fluorescent puncta size and density. / Dumitriu, Dani; Berger, Seth I.; Hamo, Carine; Hara, Yuko; Bailey, Megan; Hamo, Amarelle; Grossman, Yael S.; Janssen, William G.; Morrison, John.

In: Journal of Neuroscience Methods, Vol. 209, No. 1, 30.07.2012, p. 97-105.

Research output: Contribution to journalArticle

Dumitriu, D, Berger, SI, Hamo, C, Hara, Y, Bailey, M, Hamo, A, Grossman, YS, Janssen, WG & Morrison, J 2012, 'Vamping: Stereology-based automated quantification of fluorescent puncta size and density', Journal of Neuroscience Methods, vol. 209, no. 1, pp. 97-105. https://doi.org/10.1016/j.jneumeth.2012.05.031
Dumitriu, Dani ; Berger, Seth I. ; Hamo, Carine ; Hara, Yuko ; Bailey, Megan ; Hamo, Amarelle ; Grossman, Yael S. ; Janssen, William G. ; Morrison, John. / Vamping : Stereology-based automated quantification of fluorescent puncta size and density. In: Journal of Neuroscience Methods. 2012 ; Vol. 209, No. 1. pp. 97-105.
@article{d17ba597373d485c94c6231a2832928b,
title = "Vamping: Stereology-based automated quantification of fluorescent puncta size and density",
abstract = "The size of dendritic spines and postsynaptic densities (PSDs) is well known to be correlated with molecular and functional characteristics of the synapse. Thus, the development of microscopy methods that allow high throughput quantification and measurement of PSDs is a contemporary need in the field of neurobiology. While the gold standard for measurement of sub-micrometer structures remains electron microscopy (EM), this method is exceedingly laborious and therefore not always feasible. Immunohistochemistry (IHC) is a much faster technique for identifying biological structures such as PSDs, but the fluorescent images resulting from it have traditionally been harder to interpret and quantify. Here, we report on two new image analysis tools that result in accurate size and density measurements of fluorescent puncta. Anti-PSD-95 staining was used to target synapses. The new technique of vamping, using Volume Assisted Measurement of Puncta in 2 and 3 Dimensions (VAMP2D and VAMP3D) respectively, is based on stereological principles. The fully automated image analysis tool was tested on the same subjects for whom we had previously obtained EM measurements of PSD size and/or density. Based on highly consistent results between data obtained by each of these methods, vamping offers an expedient alternative to EM that can nonetheless deliver a high level of accuracy in measuring sub-cellular structures.",
keywords = "Confocal microscopy, Electron microscopy, Image analysis, Postsynaptic densities, PSD-95, Stereology",
author = "Dani Dumitriu and Berger, {Seth I.} and Carine Hamo and Yuko Hara and Megan Bailey and Amarelle Hamo and Grossman, {Yael S.} and Janssen, {William G.} and John Morrison",
year = "2012",
month = "7",
day = "30",
doi = "10.1016/j.jneumeth.2012.05.031",
language = "English (US)",
volume = "209",
pages = "97--105",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Vamping

T2 - Stereology-based automated quantification of fluorescent puncta size and density

AU - Dumitriu, Dani

AU - Berger, Seth I.

AU - Hamo, Carine

AU - Hara, Yuko

AU - Bailey, Megan

AU - Hamo, Amarelle

AU - Grossman, Yael S.

AU - Janssen, William G.

AU - Morrison, John

PY - 2012/7/30

Y1 - 2012/7/30

N2 - The size of dendritic spines and postsynaptic densities (PSDs) is well known to be correlated with molecular and functional characteristics of the synapse. Thus, the development of microscopy methods that allow high throughput quantification and measurement of PSDs is a contemporary need in the field of neurobiology. While the gold standard for measurement of sub-micrometer structures remains electron microscopy (EM), this method is exceedingly laborious and therefore not always feasible. Immunohistochemistry (IHC) is a much faster technique for identifying biological structures such as PSDs, but the fluorescent images resulting from it have traditionally been harder to interpret and quantify. Here, we report on two new image analysis tools that result in accurate size and density measurements of fluorescent puncta. Anti-PSD-95 staining was used to target synapses. The new technique of vamping, using Volume Assisted Measurement of Puncta in 2 and 3 Dimensions (VAMP2D and VAMP3D) respectively, is based on stereological principles. The fully automated image analysis tool was tested on the same subjects for whom we had previously obtained EM measurements of PSD size and/or density. Based on highly consistent results between data obtained by each of these methods, vamping offers an expedient alternative to EM that can nonetheless deliver a high level of accuracy in measuring sub-cellular structures.

AB - The size of dendritic spines and postsynaptic densities (PSDs) is well known to be correlated with molecular and functional characteristics of the synapse. Thus, the development of microscopy methods that allow high throughput quantification and measurement of PSDs is a contemporary need in the field of neurobiology. While the gold standard for measurement of sub-micrometer structures remains electron microscopy (EM), this method is exceedingly laborious and therefore not always feasible. Immunohistochemistry (IHC) is a much faster technique for identifying biological structures such as PSDs, but the fluorescent images resulting from it have traditionally been harder to interpret and quantify. Here, we report on two new image analysis tools that result in accurate size and density measurements of fluorescent puncta. Anti-PSD-95 staining was used to target synapses. The new technique of vamping, using Volume Assisted Measurement of Puncta in 2 and 3 Dimensions (VAMP2D and VAMP3D) respectively, is based on stereological principles. The fully automated image analysis tool was tested on the same subjects for whom we had previously obtained EM measurements of PSD size and/or density. Based on highly consistent results between data obtained by each of these methods, vamping offers an expedient alternative to EM that can nonetheless deliver a high level of accuracy in measuring sub-cellular structures.

KW - Confocal microscopy

KW - Electron microscopy

KW - Image analysis

KW - Postsynaptic densities

KW - PSD-95

KW - Stereology

UR - http://www.scopus.com/inward/record.url?scp=84863228490&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84863228490&partnerID=8YFLogxK

U2 - 10.1016/j.jneumeth.2012.05.031

DO - 10.1016/j.jneumeth.2012.05.031

M3 - Article

VL - 209

SP - 97

EP - 105

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 1

ER -