Studies on stress-induced changes at the subcellular level by Raman microspectroscopic mapping

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Christoph Krafft - , Technische Universität Dresden (Autor:in)
  • Thomas Knetschke - , Institut für Anatomie (Autor:in)
  • Richard H.W. Funk - , Institut für Anatomie (Autor:in)
  • Reiner Salzer - , Technische Universität Dresden (Autor:in)

Abstract

Raman microspectroscopic mapping enables one to study the chemical composition and molecular structure of subcellular components in individual cells without the need for labeling. Lung fibroblast cells were prepared under normal conditions and under stress, which was induced by 24 h of exposure to glyoxal. Raman microspectroscopic maps were recorded from fixed cells with 785-nm excitation and with 1-μm step width. Cluster analysis was applied to generate pseudocolor images of the cell morphology. Raman maps revealed that the cell nucleus shrinks in stressed cells, called pyknosis, which refers to an early stage of apoptosis. The intensity of nucleic acid bands decreased in cluster-averaged Raman spectra of the nucleus and cytoplasm, which is consistent with degradation and conformational changes of DNA and RNA. During a later stage of apoptosis, Raman maps indicate a rounding of cells, a further intensity decrease of nucleic acids bands, fragmentation of the nucleus, disappearance of lipid bodies, and formation of blisters at the cell surface. Whereas the peripheral membrane of the undisturbed cell is composed of lipids and cholesterol, the blisters have a higher protein content with nucleic acids incorporated. The results demonstrate that Raman spectroscopic mapping might become a powerful tool in cell biology for single cell analysis.

Details

OriginalspracheEnglisch
Seiten (von - bis)4424-4429
Seitenumfang6
FachzeitschriftAnalytical Chemistry
Jahrgang78
Ausgabenummer13
PublikationsstatusVeröffentlicht - 1 Juli 2006
Peer-Review-StatusJa

Externe IDs

PubMed 16808450

Schlagworte

ASJC Scopus Sachgebiete