A simple, straightforward correlative live-cell-imaging-structured-illumination-microscopy approach for studying organelle dynamics

Shachar Sherman, Dikla Nachmias, Natalie Elia

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Most cellular organelles are highly dynamic and continuously undergo membrane fission and fusion to mediate their function. Documenting organelle dynamics under physiological conditions, therefore, requires high temporal resolution of the recording system. Concurrently, these structures are relatively small and determining their substructural organization is often impossible using conventional microscopy. Structured Illumination Microscopy (SIM) is a super resolution technique providing a two-fold increase in resolution. Importantly, SIM is versatile because it allows the use of any fluorescent dye or protein and, hence, is highly applicable for cell biology. However, similar to other SR techniques, the applicability of SIM to high-speed live cell imaging is limited. Here we present an easy, straightforward methodology for coupling of high-speed live cell recordings, using spinning disk (SD) microscopy, with SIM. Using this simple methodology, we are able to track individual mitochondrial membrane fission and fusion events in real time and to determine the network connectivity and substructural organization of the membrane at high resolution. Applying this methodology to other cellular organelles such as, ER, golgi, and cilia will no doubt contribute to our understanding of membrane dynamics in cells.

Original languageEnglish
Pages (from-to)777-783
Number of pages7
JournalMicroscopy Research and Technique
Volume78
Issue number9
DOIs
StatePublished - 1 Sep 2015

Keywords

  • Fission
  • Fusion
  • Light microscopy
  • Membrane dynamics
  • Mitochondria
  • SIM
  • Spinning Disk

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Instrumentation
  • Medical Laboratory Technology

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