Co-registration of total internal reflection fluorescence and confocal microscopy images for studying vesicle trafficking in living cells

William T E Pitkeathly*, Joshua Z. Rappoport, Ela Claridge

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Scopus citations

Abstract

The dynamics of vesicle trafficking via the cell membrane and the tracking of their subsequent movements within the cell are of great interest in the biomedical sciences. One challenge of image analysis is following the vesicle's fate continuously from its formation at the membrane to its final destination, as it requires different microscopy techniques to image the complete journey. Total internal reflection fluorescence (TIRF) microscopy is used for imaging events at the cell membrane and laser scanning confocal microscopy (LSCM) is used for imaging the interior of the cell. We present a simple and robust method for co-registration of data sets from the two microscopy techniques. This method is validated on images generated by computer simulation of the image formation process in TIRF and in LSCM. The registration parameters are recovered with error less than 1% in presence of Gaussian noise up to SNR of 3.8dB. Registration of real microscopy data is shown and the accuracy of the retrieved parameters is compared and agrees well with values obtained manually and a difference in squares measure.

Original languageEnglish (US)
Title of host publication2012 9th IEEE International Symposium on Biomedical Imaging
Subtitle of host publicationFrom Nano to Macro, ISBI 2012 - Proceedings
Pages182-185
Number of pages4
DOIs
StatePublished - 2012
Event2012 9th IEEE International Symposium on Biomedical Imaging: From Nano to Macro, ISBI 2012 - Barcelona, Spain
Duration: May 2 2012May 5 2012

Publication series

NameProceedings - International Symposium on Biomedical Imaging
ISSN (Print)1945-7928
ISSN (Electronic)1945-8452

Other

Other2012 9th IEEE International Symposium on Biomedical Imaging: From Nano to Macro, ISBI 2012
CountrySpain
CityBarcelona
Period5/2/125/5/12

Keywords

  • TIRF
  • confocal
  • fluorescence microscopy
  • multi-modality
  • registration

ASJC Scopus subject areas

  • Biomedical Engineering
  • Radiology Nuclear Medicine and imaging

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