Spectral Fiber-Photometry Derives Hemoglobin-Absorption Changes for Accurate Measurement of Fluorescent Sensor Activity

2021 ◽  
Author(s):  
Wei-Ting Zhang ◽  
Tzu-Hao Harry Chao ◽  
Yue Yang ◽  
Tzu-Wen Wang ◽  
Sung-Ho Lee ◽  
...  
Keyword(s):  
Fluorescent Sensor ◽  
Hemoglobin Absorption

scholarly journals Spectral fiber-photometry derives hemoglobin-absorption changes for accurate measurement of fluorescent sensors

2021 ◽  
Author(s):  
Wei-Ting Zhang ◽  
Tzu-Hao Harry Chao ◽  
Yue Yang ◽  
Tzu-Wen Wang ◽  
Sung-Ho Lee ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Transfer Functions ◽  
Fluorescent Sensor ◽  
Hemoglobin Concentration ◽  
Computational Method ◽  
Photon Absorption ◽  
Concentration Changes ◽  
Spectral Photometry ◽  
The Brain ◽  
Hemoglobin Absorption

Fiber-photometry is an emerging technique for recording fluorescent sensor activity in the brain. Here we demonstrate significant hemoglobin-absorption artifacts that may be misinterpreted as sensor activity changes. Because hemoglobin exists in nearly every location in the brain and its concentration varies over time, such artifacts could significantly impede the accuracy of photometry recording results. We present a novel use of spectral photometry technique and propose a computational method to quantify photon absorption effect from an activity-independent fluorescent protein, which can be used to derive oxy- and deoxy-hemoglobin concentration changes and correct target sensor activities across spectra. We further demonstrate the utility of this method for delineating brain regional differences in neurovascular transfer functions and shed light on the interpretation of hemodynamic-based neuroimaging data.

Single-Molecule Imaging of Active Mitochondrial Nitroreductases using a Photo-Crosslinking Fluorescent Sensor

2019 ◽  
Author(s):  
Zacharias Thiel ◽  
Pablo Rivera-Fuentes
Keyword(s):  
Subcellular Localization ◽  
Fluorescent Probe ◽  
Single Molecule ◽  
Mammalian Cells ◽  
Fluorescent Sensor ◽  
Biological Functions ◽  
Localization Microscopy ◽  
Drug Activation ◽  
Nitroreductase Activity ◽  
Single Molecule Localization Microscopy

Many biomacromolecules are known to cluster in microdomains with specific subcellular localization. In the case of enzymes, this clustering greatly defines their biological functions. Nitroreductases are enzymes capable of reducing nitro groups to amines and play a role in detoxification and pro-drug activation. Although nitroreductase activity has been detected in mammalian cells, the subcellular localization of this activity remains incompletely characterized. Here, we report a fluorescent probe that enables super-resolved imaging of pools of nitroreductase activity within mitochondria. This probe is activated sequentially by nitroreductases and light to give a photo-crosslinked adduct of active enzymes. In combination with a general photoactivatable marker of mitochondria, we performed two-color, threedimensional, single-molecule localization microscopy. These experiments allowed us to image the sub-mitochondrial organization of microdomains of nitroreductase activity.<br>

Single-Molecule Imaging of Active Mitochondrial Nitroreductases using a Photo-Crosslinking Fluorescent Sensor

2019 ◽  
Author(s):  
Zacharias Thiel ◽  
Pablo Rivera-Fuentes
Keyword(s):  
Subcellular Localization ◽  
Fluorescent Probe ◽  
Single Molecule ◽  
Mammalian Cells ◽  
Fluorescent Sensor ◽  
Biological Functions ◽  
Localization Microscopy ◽  
Drug Activation ◽  
Nitroreductase Activity ◽  
Single Molecule Localization Microscopy

Many biomacromolecules are known to cluster in microdomains with specific subcellular localization. In the case of enzymes, this clustering greatly defines their biological functions. Nitroreductases are enzymes capable of reducing nitro groups to amines and play a role in detoxification and pro-drug activation. Although nitroreductase activity has been detected in mammalian cells, the subcellular localization of this activity remains incompletely characterized. Here, we report a fluorescent probe that enables super-resolved imaging of pools of nitroreductase activity within mitochondria. This probe is activated sequentially by nitroreductases and light to give a photo-crosslinked adduct of active enzymes. In combination with a general photoactivatable marker of mitochondria, we performed two-color, threedimensional, single-molecule localization microscopy. These experiments allowed us to image the sub-mitochondrial organization of microdomains of nitroreductase activity.<br>

A New Fluorescent Salen-uranyl Sensor for the Sub-ppm Detection of Chemical Warfare Agents

2020 ◽  
Vol 24 (20) ◽  
pp. 2378-2382
Author(s):  
Andrea Pappalardo ◽  
Chiara M.A. Gangemi ◽  
Rosa Maria Toscano ◽  
Giuseppe Trusso Sfrazzetto
Keyword(s):  
Real Time ◽  
Fluorescent Sensor ◽  
Chemical Warfare Agents ◽  
Optical Response ◽  
Chemical Warfare ◽  
Lethal Effects ◽  
Strong Affinity ◽  
Chemical Attack ◽  
Fluorescence Measurements ◽  
Warfare Agents

Real-time sensing of Chemical Warfare Agents (CWAs) is today a crucial topic to prevent the lethal effects of a terroristic chemical attack. For this reason, the development of efficient, selective, sensitive and reversible sensoristic devices, able to detect by optical response ppm levels of these compounds, is strongly required. Here, the synthesis of a new fluorescent sensor based on a salen-uranyl scaffold, functionalized with two bodipy moieties, and its application for the detection of sub-ppm levels of CWAs is reported. Detection properties were evaluated by fluorescence measurements and selectivity tests demonstrated the strong affinity for CWAs.

Lanthanide Metal–Organic Framework-Based Fluorescent Sensor Arrays to Discriminate and Quantify Ingredients of Natural Medicine

Langmuir ◽  
2021 ◽  
Vol 37 (17) ◽  
pp. 5321-5328
Author(s):  
Kunpeng Yin ◽  
Siqi Wu ◽  
Hao Zheng ◽  
Liang Gao ◽  
Jinfeng Liu ◽  
...  
Keyword(s):  
Fluorescent Sensor ◽  
Metal Organic Framework ◽  
Sensor Arrays ◽  
Natural Medicine ◽  
Metal Organic ◽  
Lanthanide Metal ◽  
Organic Framework

Fluorescent sensor based on triphenylamine for Zn2+ with high selectivity and imaging in living cells

2021 ◽  
Vol 251 ◽  
pp. 119480
Author(s):  
Dongyuan Wang ◽  
Qiang Yin ◽  
Ming Zheng ◽  
Yongbo Xie ◽  
Wen He ◽  
...  
Keyword(s):  
High Selectivity ◽  
Fluorescent Sensor ◽  
Living Cells
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