Computed tomography-based spectral imaging for fluorescence microscopy

Bridget K. Ford, Curtis E. Volin, Sean M. Murphy, Ronald M. Lynch, Michael R. Descour

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

The computed tomography imaging spectrometer (CTIS) is a non-scanning instrument capable of simultaneously acquiring full spectral information (450-750 nm) from every position element within its field of view (75 μm × 75 μm). The current spatial and spectral sampling intervals of the spectrometer are 1.0 μm and 10 nm, respectively. This level of resolution is adequate to resolve signal responses from multiple fluorescence probes located within individual cells or different locations within the same cell. Spectral imaging results are presented from the CTIS combined with a commercial inverted fluorescence microscope. Results demonstrate the capability of the CTIS to monitor the spatiotemporal evolution of pH in rat insulinoma cells loaded with SNARF-1. The ability to analyze full spectral information for two-dimensional (x, y) images allows precise evaluation of heterogeneous physiological responses within cell populations. Due to low signal levels, integration times up to 2 s were required. However, reasonable modifications to the instrument design will provide higher system transmission efficiency with increased temporal and spatial resolution. Specifically, a custom optical design including the use of a larger format detector array is under development for a second-generation system.

Original languageEnglish (US)
Pages (from-to)986-993
Number of pages8
JournalBiophysical Journal
Volume80
Issue number2
DOIs
StatePublished - 2001

ASJC Scopus subject areas

  • Biophysics

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