Total internal reflection microscopy for live imaging of cellular uptake of sub-micron non-fluorescent particles

G. D. Byrne, M. C. Pitter, J. Zhang, F. H. Falcone, S. Stolnik, Michael Geoffrey Somekh

Research output: Journal article publicationJournal articleAcademic researchpeer-review

34 Citations (Scopus)


This paper presents a simple, high-resolution, non-fluorescent imaging technique called total internal reflection microscopy (TIRM) and demonstrates its potential application to real-time imaging of live cellular events. In addition, a novel instrument is introduced that combines the simplicity of TIRM with the specificity afforded by dual-colour total internal reflection fluorescence (TIRF) microscopy and allows sequential imaging with the two modalities. The key design considerations necessary to apply these imaging modes in a single instrument are discussed. The application of TIRM alone yielded high-resolution live images of cell adherence to a poly-l-lysine modified substrate, whereby fine cellular structures are imaged. Non-fluorescent imaging of the uptake of sub-micron-sized polymeric particles by live cells is also demonstrated. Finally, images of fluorescently labelled cells were obtained in TIRF mode, sequentially to images obtained of the same cell in TIRM mode. Visual information gained using TIRF is compared with TIRM to demonstrate that the level of cell structure information obtainable with our total internal reflection microscope is comparable with the TIRF technique.
Original languageEnglish
Pages (from-to)168-179
Number of pages12
JournalJournal of Microscopy
Issue number1
Publication statusPublished - 1 Jul 2008
Externally publishedYes


  • Drug delivery
  • Endocytosis
  • Evanescent wave microscopy
  • Imaging systems
  • Live-cell imaging
  • Medical and biological imaging
  • Microscopy
  • Total internal reflection fluorescence

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

Cite this