UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator.

Foote, JR; Levine, AP; Behe, P; Duchen, MR; Segal, AW; (2017) Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator. Journal of Visualized Experiments (122) , Article e55107. 10.3791/55107. Green open access

[img]
Preview
Text
Levine_jove-protocol-55107-imaging-neutrophil-phagosome-cytoplasm-using-ratiometric-ph.pdf - Published version

Download (777kB) | Preview

Abstract

Neutrophils are crucial to host innate defense and, consequently, constitute an important area of medical research. The phagosome, the intracellular compartment where the killing and digestion of engulfed particles take place, is the main arena for neutrophil pathogen killing that requires tight regulation. Phagosomal pH is one aspect that is carefully controlled, in turn regulating antimicrobial protease activity. Many fluorescent pH-sensitive dyes have been used to visualize the phagosomal environment. S-1 has several advantages over other pH-sensitive dyes, including its dual emission spectra, its resistance to photo-bleaching, and its high pKa. Using this method, we have demonstrated that the neutrophil phagosome is unusually alkaline in comparison to other phagocytes. By using different biochemical conjugations of the dye, the phagosome can be delineated from the cytoplasm so that changes in the size and shape of the phagosome can be assessed. This allows for further monitoring of ionic movement.

Type: Article
Title: Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator.
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.3791/55107
Publisher version: http://dx.doi.org/10.3791/55107
Language: English
Additional information: Copyright © 2017 Creative Commons Attribution 3.0 License
Keywords: Immunology, phagocytosis, neutrophil, phagosomal pH, ratiometric imaging, fluorescence microscopy, innate immunity
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Pathology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
URI: https://discovery.ucl.ac.uk/id/eprint/1555302
Downloads since deposit
45Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item