Share Email Print

Proceedings Paper

Sensitive imaging of spectrally overlapping flourochromes using the LSM 510 META
Author(s): Mary E. Dickinson; Christopher W Waters; Gregory H. Bearman; Ralf Wolleschensky; Sebastian Tille; Scott E. Fraser
Format Member Price Non-Member Price
PDF $14.40 $18.00
cover GOOD NEWS! Your organization subscribes to the SPIE Digital Library. You may be able to download this paper for free. Check Access

Paper Abstract

Multi-color fluorescence microscopy has become a popular way to discriminate between multiple proteins, organelles or functions in a single cell or animal and can be used to approximate the physical relationships between individual proteins within the cell, for instance, by using Fluorescence Resonance Energy Transfer (FRET). However, as researchers attempt to gain more information from single samples by using multiple dyes or fluorescent proteins (FPs), spectral overlap between emission signals can obscure the data. Signal separation using glass filters is often impractical for many dye combinations. In cases where there is extensive overlap between fluorochromes, separation is often physically impossible or can only be achieved by sacrificing signal intensity. Here we test the performance of a new, integrated laser scanning system for multispectral imaging, the Zeiss LSM 510 META. This system consists of a sensitive multispectral imager and online linear unmixing functions integrated into the system software. Below we describe the design of the META device and show results from tests of the linear unmixing experiments using fluorochromes with overlapping emission spectra. These studies show that it is possible to expand the number of dyes used in multicolor applications.

Paper Details

Date Published: 17 June 2002
PDF: 14 pages
Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002); doi: 10.1117/12.470686
Show Author Affiliations
Mary E. Dickinson, Beckman Laser Institute and Medical Ctr. (United States)
Christopher W Waters, Beckman Laser Institute and Medical Ctr. (United States)
Gregory H. Bearman, Jet Propulsion Lab (United States)
Ralf Wolleschensky, Carl Zeiss Jena GmbH (Germany)
Sebastian Tille, Carl Zeiss MicroImaging, Inc. (United States)
Scott E. Fraser, Beckman Laser Institute and Medical Ctr. (United States)

Published in SPIE Proceedings Vol. 4620:
Multiphoton Microscopy in the Biomedical Sciences II
Ammasi Periasamy; Peter T. C. So, Editor(s)

© SPIE. Terms of Use
Back to Top