Microglia pp 261-279 | Cite as

Tomato Lectin Histochemistry for Microglial Visualization

  • Nàdia Villacampa
  • Beatriz Almolda
  • Berta González
  • Bernardo Castellano
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1041)

Abstract

The use of different lectins for the study of microglial cells in the central nervous system (CNS) is a valuable tool that has been extensively used in the last years for the selective staining of this glial cell population, not only in normal physiological conditions, but also in a wide range of pathological situations where the normal homeostasis of the parenchyma is disturbed. In this chapter we accurately describe the methodology for the selective labelling of microglial cells by using the tomato lectin (TL), a protein lectin obtained from Lycopersicum esculentum with specific affinity for poly-N-acetyl lactosamine sugar residues which are found on the plasma membrane and in the cytoplasm of microglia. Here we describe how to perform this technique on vibratome, frozen, and paraffin sections for optical microscopy, as well as for transmission electron microscopy (TEM) studies. Using this methodology it is possible to visualize amoeboid microglia in the developing brain, ramified microglia in the adult, and activated/reactive microglia in the experimentally damaged brain. In addition, as TL also recognized sugar residues in endothelial cells, this technique is very useful for the study of the relationship established between microglia and the CNS vasculature.

Key words

Microglia Histochemistry Tomato lectin Transmission electron microscopy Double labelling Fluorescence 

Abbreviations

AAS

Acetic acid solution

BBS

Buffer blocking solution

CNS

Central nervous system

DS

Developing solution

DSN

Developing solution with nickel

DSNC

Developing solution with nickel-cobalt-enhanced DAB

EPBS

Endogenous peroxidase blocking solution

FS

Fixative solution

FSG

Fixative solution with glutaraldehyde

GFAP

Glial fibrillary acidic protein

GSA-IB4

Griffonia simplicifolia B4 isolectin

MHC

Major histocompatibility complex

ML-1

Mistletoe lectin

OS

Olmos solution

OTS

Osmium tetroxide solution

PBS

Phosphate buffer solution

PWB

Phosphate washing buffer

RCA-1

Ricinus communis agglutinin-1

SAS

Sodium acetate solution

SS

Sucrose solution

TB

Tris-buffered solution

TBS

Tris-buffered saline solution

TBST

TBS with Triton X-100

TEM

Transmission electron microscopy

TL

Tomato lectin

UAS

Uranyl acetate solution

WB

Walpole buffer

WGA

Wheat germ agglutinin

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Nàdia Villacampa
    • 1
  • Beatriz Almolda
    • 1
  • Berta González
    • 1
  • Bernardo Castellano
    • 2
  1. 1.Universitat Autònoma de BarcelonaBarcelonaSpain
  2. 2.Universitat Autònoma de BarcelonaBellaterraSpain

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