Skip to main content
Article thumbnail
Location of Repository

Translocation or just location? Pseudopodia affect fluorescent signals

By Sharon Dewitt, Richard L. Darley and Maurice B. Hallett

Abstract

The use of fluorescent probes is one of the most powerful techniques for gaining spatial and temporal knowledge of dynamic events within living cells. Localized increases in the signal from cytosolic fluorescent protein constructs, for example, are frequently used as evidence for translocation of proteins to specific sites within the cell. However, differences in optical and geometrical properties of cytoplasm can influence the recorded intensity of the probe signal. Pseudopodia are especially problematic because their cytoplasmic properties can cause abrupt increases in fluorescent signal of both GFP and fluorescein. Investigators should therefore be cautious when interpreting fluorescence changes within a cell, as these can result from either translocation of the probe or changes in the optical properties of the milieu surrounding the probe

Topics: News
Publisher: The Rockefeller University Press
OAI identifier: oai:pubmedcentral.nih.gov:2654297
Provided by: PubMed Central
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.pubmedcentral.nih.g... (external link)
  • Suggested articles

    Citations

    1. (2005). A guide to choosing fl uorescent proteins.
    2. (1991). Ca 2+ gradients underlying polarization and chemotaxis of eosinophils.
    3. (1995). Chemoattractantcontrolled accumulation of coronin at the leading edge of Dictyostelium cells monitored using a green fl uorescent protein-coronin fusion protein.
    4. (1908). Contributions to the optics of turbid media particularly of colloidal metal solutions
    5. (2002). Cytosolic free Ca 2+ changes and calpain activation are required for 2 integrin-accelerated phagocytosis by human neutrophils.
    6. (1995). Does cytosolic free Ca 2+ signal neutrophil chemotaxis?
    7. (1989). Fluorescent probes of cell signalling.
    8. (1985). Intracellular free calcium localization in neutrophils during phagocytosis.
    9. (2007). Ironing out the wrinkles of neutrophil phagocytosis: membrane reservoirs for surface area expansion.
    10. (1979). Light scattering from biological cells: dependence of backscatter radiation on membrane thickness and refractive index.
    11. (2002). Lipid products of PI(3)Ks maintain persistent cell polarity and directed motility in neutrophils.
    12. (1997). Live dynamics of Dictyostelium cofi lin suggests a role in remodeling actin latticework into bundles.
    13. (1973). Living Blood Cells and their Ultrastructure Springer-Verlag,
    14. (2006). Localised PtdIns(3,4,5)P-3 or PtdIns(3,4)P-2 at the phagocytic cup is required for both phagosome closure and
    15. (2002). Measurement of cytosolic free Ca 2+ in human neutrophils.
    16. (2007). Mie Scattering calculator. http:// omlc.ogi.edu/calc/mie_calc.html Ruban
    17. (1980). Morphometry of human leukocytes.
    18. (1990). Oxidase activation in individual neutrophils is dependent on the onset and magnitude of the Ca 2+ signal .
    19. (2000). Polarization of chemoattractant receptor signaling during neutrophil chemotaxis.
    20. (2015). Probing the structure of cytoplasm.
    21. (2006). Re-assembly of a contractile actin cortex in cell blebs.
    22. (2008). SHIP-1 increases early oxidative burst and regulates phagosome maturation in macrophages .

    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.