Simultaneous detection of multiple targets for ultrastructural immunocytochemistry.

Publisher: Springer Country of Publication: Germany NLM ID: 9506663 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-119X (Electronic) Linking ISSN: 09486143 NLM ISO Abbreviation: Histochem Cell Biol Subsets: MEDLINE

Original Publication: Berlin : Springer,

Immunohistochemistry/*methods ; Metal Nanoparticles/*chemistry ; Staining and Labeling/*methods; Actins/metabolism ; Antibodies/immunology ; Carrier Proteins/metabolism ; Cell Cycle Proteins ; Cell Line, Tumor ; Cell Nucleus ; Gold/chemistry ; HeLa Cells ; Humans ; Microscopy, Electron ; Nuclear Proteins/metabolism ; Nucleophosmin ; Phosphatidylinositol 4,5-Diphosphate/metabolism ; Ribonucleoproteins, Small Nuclear/metabolism

Simultaneous detection of biological molecules by means of indirect immunolabeling provides valuable information about their localization in cellular compartments and their possible interactions in macromolecular complexes. While fluorescent microscopy allows for simultaneous detection of multiple antigens, the sensitive electron microscopy immunodetection is limited to only two antigens. In order to overcome this limitation, we prepared a set of novel, shape-coded metal nanoparticles readily discernible in transmission electron microscopy which can be conjugated to antibodies or other bioreactive molecules. With the use of novel nanoparticles, various combinations with commercial gold nanoparticles can be made to obtain a set for simultaneous labeling. For the first time in ultrastructural histochemistry, up to five molecular targets can be identified simultaneously. We demonstrate the usefulness of the method by mapping of the localization of nuclear lipid phosphatidylinositol-4,5-bisphosphate together with four other molecules crucial for genome function, which proves its suitability for a wide range of biomedical applications.

Erratum in: Histochem Cell Biol. 2014 Mar;141(3):241.

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0 (Actins)
0 (Antibodies)
0 (Carrier Proteins)
0 (Cell Cycle Proteins)
0 (Nuclear Proteins)
0 (Phosphatidylinositol 4,5-Diphosphate)
0 (Ribonucleoproteins, Small Nuclear)
0 (SMC2 protein, human)
117896-08-9 (Nucleophosmin)
7440-57-5 (Gold)

Date Created: 20140123 Date Completed: 20141118 Latest Revision: 20211203

20221213

PMC3935117

10.1007/s00418-013-1178-6

24449180