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Novel approach for accurate tissue-based protein colocalization and proximity microscopy.

ABSTRACT: Fluorescence colocalization microscopy is frequently used to assess potential links between distinct molecules; however, this method can lead to striking false-positive results and erroneous conclusions. Here we developed a novel approach with more sophisticated mathematical colocalization analyses together with visualization of physical proximity using fluorescence resonance energy transfer (FRET). To verify our results we used the proximity ligation assay (PLA). With these methods we could demonstrate that distinct neurodegeneration-related proteins either not or only rarely interact in human brain tissue.

SUBMITTER: Lutz MI

PROVIDER: S-EPMC5454019 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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Novel approach for accurate tissue-based protein colocalization and proximity microscopy.

Lutz Mirjam I MI Schwaiger Carmen C Hochreiter Bernhard B Kovacs Gabor G GG Schmid Johannes A JA

Scientific reports 20170601 1

Fluorescence colocalization microscopy is frequently used to assess potential links between distinct molecules; however, this method can lead to striking false-positive results and erroneous conclusions. Here we developed a novel approach with more sophisticated mathematical colocalization analyses together with visualization of physical proximity using fluorescence resonance energy transfer (FRET). To verify our results we used the proximity ligation assay (PLA). With these methods we could dem ...[more]

PMID: 28572629

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