scholarly journals Quantum Dot–Fluorescent Protein FRET Probes for Sensing Intracellular pH

ACS Nano ◽  
2012 ◽  
Vol 6 (4) ◽  
pp. 2917-2924 ◽  
Author(s):  
Allison M. Dennis ◽  
Won Jong Rhee ◽  
David Sotto ◽  
Steven N. Dublin ◽  
Gang Bao
Keyword(s):  
Quantum Dot ◽  
Intracellular Ph ◽  
Fluorescent Protein

scholarly journals Green Fluorescent Protein as a Noninvasive Intracellular pH Indicator

1998 ◽  
Vol 74 (3) ◽  
pp. 1591-1599 ◽  
Author(s):  
Malea Kneen ◽  
Javier Farinas ◽  
Yuxin Li ◽  
A.S. Verkman
Keyword(s):  
Green Fluorescent Protein ◽  
Intracellular Ph ◽  
Fluorescent Protein ◽  
Ph Indicator ◽  
Green Fluorescent

scholarly journals In Vivo Indicators of Cytoplasmic, Vacuolar, and Extracellular pH Using pHluorin2 in Candida albicans

mSphere ◽  
2017 ◽  
Vol 2 (4) ◽  
Author(s):  
Hélène Tournu ◽  
Arturo Luna-Tapia ◽  
Brian M. Peters ◽  
Glen E. Palmer
Keyword(s):  
Candida Albicans ◽  
Intracellular Ph ◽  
Fungal Pathogen ◽  
Fluorescent Protein ◽  
Extracellular Ph ◽  
Adaptive Responses ◽  
Ph Homeostasis ◽  
Content Type ◽  
A Cell

ABSTRACT Candida albicans is an opportunistic fungal pathogen that colonizes the reproductive and gastrointestinal tracts of its human host. It can also invade the bloodstream and deeper organs of immunosuppressed individuals, and thus it encounters enormous variations in external pH in vivo. Accordingly, survival within such diverse niches necessitates robust adaptive responses to regulate intracellular pH. However, the impact of antifungal drugs upon these adaptive responses, and on intracellular pH in general, is not well characterized. Furthermore, the tools and methods currently available to directly monitor intracellular pH in C. albicans, as well as other fungal pathogens, have significant limitations. To address these issues, we developed a new and improved set of pH sensors based on the pH-responsive fluorescent protein pHluorin. This includes a cytoplasmic sensor, a probe that localizes inside the fungal vacuole (an acidified compartment that plays a central role in intracellular pH homeostasis), and a cell surface probe that can detect changes in extracellular pH. These tools can be used to monitor pH within single C. albicans cells or in cell populations in real time through convenient and high-throughput assays. Environmental or chemically induced stresses often trigger physiological responses that regulate intracellular pH. As such, the capacity to detect pH changes in real time and within live cells is of fundamental importance to essentially all aspects of biology. In this respect, pHluorin, a pH-sensitive variant of green fluorescent protein, has provided an invaluable tool to detect such responses. Here, we report the adaptation of pHluorin2 (PHL2), a substantially brighter variant of pHluorin, for use with the human fungal pathogen Candida albicans. As well as a cytoplasmic PHL2 indicator, we describe a version that specifically localizes within the fungal vacuole, an acidified subcellular compartment with important functions in nutrient storage and pH homeostasis. In addition, by means of a glycophosphatidylinositol-anchored PHL2-fusion protein, we generated a cell surface pH sensor. We demonstrated the utility of these tools in several applications, including accurate intracellular and extracellular pH measurements in individual cells via flow cytometry and in cell populations via a convenient plate reader-based protocol. The PHL2 tools can also be used for endpoint as well as time course experiments and to conduct chemical screens to identify drugs that alter normal pH homeostasis. These tools enable observation of the highly dynamic intracellular pH shifts that occur throughout the fungal growth cycle, as well as in response to various chemical treatments. IMPORTANCE Candida albicans is an opportunistic fungal pathogen that colonizes the reproductive and gastrointestinal tracts of its human host. It can also invade the bloodstream and deeper organs of immunosuppressed individuals, and thus it encounters enormous variations in external pH in vivo. Accordingly, survival within such diverse niches necessitates robust adaptive responses to regulate intracellular pH. However, the impact of antifungal drugs upon these adaptive responses, and on intracellular pH in general, is not well characterized. Furthermore, the tools and methods currently available to directly monitor intracellular pH in C. albicans, as well as other fungal pathogens, have significant limitations. To address these issues, we developed a new and improved set of pH sensors based on the pH-responsive fluorescent protein pHluorin. This includes a cytoplasmic sensor, a probe that localizes inside the fungal vacuole (an acidified compartment that plays a central role in intracellular pH homeostasis), and a cell surface probe that can detect changes in extracellular pH. These tools can be used to monitor pH within single C. albicans cells or in cell populations in real time through convenient and high-throughput assays.

pH-Dependent Fluorescence of a Heterologously ExpressedAequoreaGreen Fluorescent Protein Mutant: In Situ Spectral Characteristics and Applicability to Intracellular pH Estimation†,‡

Biochemistry ◽  
1998 ◽  
Vol 37 (28) ◽  
pp. 9894-9901 ◽  
Author(s):  
R. Brooks Robey ◽  
Ofelia Ruiz ◽  
Anna V. P. Santos ◽  
Jianfei Ma ◽  
Fely Kear ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Fluorescent Protein ◽  
Spectral Characteristics ◽  
Ph Dependent

scholarly journals Novel cytocompatible intracellular pH-imaging fluorescence probe composed of quantum dot and phospholipid polymer

2010 ◽  
Vol 35 (1) ◽  
pp. 147-150 ◽  
Author(s):  
Kouichi Masuda ◽  
Ryosuke Matsuno ◽  
Tomohiro Konno ◽  
Madoka Takai ◽  
Kazuhiko Ishihara
Keyword(s):  
Quantum Dot ◽  
Intracellular Ph ◽  
Fluorescence Probe ◽  
Ph Imaging ◽  
Phospholipid Polymer

Assembly and intracellular delivery of quantum dot-fluorescent protein bioconjugates

2008 ◽  
Author(s):  
Igor L. Medintz ◽  
Thomas Pons ◽  
James B. Delehanty ◽  
Kimihiro Susumu ◽  
Philip E. Dawson ◽  
...  
Keyword(s):  
Quantum Dot ◽  
Fluorescent Protein ◽  
Intracellular Delivery

scholarly journals Intracellular pH measurements made simple by fluorescent protein probes and the phasor approach to fluorescence lifetime imaging

2012 ◽  
Vol 48 (42) ◽  
pp. 5127 ◽  
Author(s):  
Antonella Battisti ◽  
Michelle A. Digman ◽  
Enrico Gratton ◽  
Barbara Storti ◽  
Fabio Beltram ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Fluorescence Lifetime ◽  
Fluorescent Protein ◽  
Fluorescence Lifetime Imaging ◽  
Ph Measurements ◽  
Lifetime Imaging
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