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Direct observation of mobility state transitions in RNA trajectories by sensitive single molecule feedback tracking.

Abstract：

:Observation and tracking of fluorescently labeled molecules and particles in living cells reveals detailed information about intracellular processes on the molecular level. Whereas light microscopic particle observation is usually limited to two-dimensional projections of short trajectory segments, we report here image-based real-time three-dimensional single particle tracking in an active feedback loop with single molecule sensitivity. We tracked particles carrying only 1-3 fluorophores deep inside living tissue with high spatio-temporal resolution. Using this approach, we succeeded to acquire trajectories containing several hundred localizations. We present statistical methods to find significant deviations from random Brownian motion in such trajectories. The analysis allowed us to directly observe transitions in the mobility of ribosomal (r)RNA and Balbiani ring (BR) messenger (m)RNA particles in living Chironomus tentans salivary gland cell nuclei. We found that BR mRNA particles displayed phases of reduced mobility, while rRNA particles showed distinct binding events in and near nucleoli.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Spille JH,Kaminski TP,Scherer K,Rinne JS,Heckel A,Kubitscheck U

doi

10.1093/nar/gku1194

subject

Has Abstract

pub_date

2015-01-01 00:00:00

pages

e14

issue

2

eissn

0305-1048

issn

1362-4962

pii

gku1194

journal_volume

43

pub_type

杂志文章

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