Bioenergetics Core
The Bioenergetics Core delivers state-of-the-art instrumentation for in-depth mitochondrial and cellular metabolic analysis. Its integrated platforms support high-throughput extracellular flux analysis, high-resolution respirometry, fluorescence imaging, and advanced mitochondrial functional assays. This Core enables investigators to assess cellular respiration, substrate utilization, mitochondrial efficiency, and overall bioenergetic health with precision and scalability.
Together with the Metabolomics Core, we provide a robust, synergistic infrastructure that supports mechanistic, translational, and systems-level studies of metabolism and mitochondrial biology in health and disease.
Core Services
Agilent Seahorse XF Metabolic Flux Analyzers
The Core houses multiple Agilent Seahorse XF platforms to measure real-time cellular bioenergetics through extracellular flux analysis. These systems quantify Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) to assess mitochondrial respiration and glycolytic function.
Available Seahorse Instruments:
- Seahorse XF Pro Analyzer – Advanced high-throughput platform with enhanced sensitivity and automated workflow capabilities.
- Seahorse XF96 Analyzer – High-throughput system for large experimental designs and drug screening studies.
- Seahorse XF24 Analyzer – Mid-throughput platform ideal for mechanistic and optimization studies.
Seahorse Assay Capabilities:
- Mito Stress Test
- Glycolysis Stress Test
- ATP Rate Assay
- Mito Fuel Flex Test
- Fatty acid oxidation assays
- ETC Enzyme Complex Activities
Sample Types Supported:
- Isolated mitochondria
- Intact living cells
- Primary cells
- Tissue homogenate
Seahorse analysis enables determination of ATP production, coupling efficiency, spare respiratory capacity, proton leak, and glycolytic reserve. The platform supports studies using immortalized cell lines, primary cells, immune cells, tissue punches, and isolated mitochondria.
Agilent Seahorse XF Metabolic Flux Analyzers
The Core houses multiple Agilent Seahorse XF platforms to measure real-time cellular bioenergetics through extracellular flux analysis. These systems quantify Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) to assess mitochondrial respiration and glycolytic function.
Available Seahorse Instruments:
- Seahorse XF Pro Analyzer – Advanced high-throughput platform with enhanced sensitivity and automated workflow capabilities.
- Seahorse XF96 Analyzer – High-throughput system for large experimental designs and drug screening studies.
- Seahorse XF24 Analyzer – Mid-throughput platform ideal for mechanistic and optimization studies.
Seahorse Assay Capabilities:
- Mito Stress Test
- Glycolysis Stress Test
- ATP Rate Assay
- Mito Fuel Flex Test
- Fatty acid oxidation assays
- ETC Enzyme Complex Activities
Sample Types Supported:
- Isolated mitochondria
- Intact living cells
- Primary cells
- Tissue homogenate
Seahorse analysis enables determination of ATP production, coupling efficiency, spare respiratory capacity, proton leak, and glycolytic reserve. The platform supports studies using immortalized cell lines, primary cells, immune cells, tissue punches, and isolated mitochondria.
BioTek Cytation 5 Multimode Reader
The BioTek Cytation 5 is a versatile multimode plate reader and automated imaging system that integrates high-throughput plate-based assays with high-resolution fluorescence microscopy.
Detection Capabilities:
- UV-Vis absorbance
- Fluorescence detection
- Luminescence detection
- Fluorescence microscopy up to 60× magnification
This platform enables quantitative plate-based measurements and immunocytochemical/immunohistological imaging within the same system.
The Cytation 5 is linked to the Seahorse platform for automated post-assay cell number normalization, improving data accuracy and reproducibility. The system operates with Gen5 software for streamlined data acquisition and analysis.
BioTek Cytation 5 Multimode Reader
The BioTek Cytation 5 is a versatile multimode plate reader and automated imaging system that integrates high-throughput plate-based assays with high-resolution fluorescence microscopy.
Detection Capabilities:
- UV-Vis absorbance
- Fluorescence detection
- Luminescence detection
- Fluorescence microscopy up to 60× magnification
This platform enables quantitative plate-based measurements and immunocytochemical/immunohistological imaging within the same system.
The Cytation 5 is linked to the Seahorse platform for automated post-assay cell number normalization, improving data accuracy and reproducibility. The system operates with Gen5 software for streamlined data acquisition and analysis.
Oroboros O2k High-Resolution Respirometry (HRR) System
The Oroboros O2k-Oxygraph provides high-resolution respirometry (HRR) coupled with high-resolution fluorometry for precise mitochondrial functional analysis.
The system utilizes dual Clark-type oxygen electrode chambers and fluorescent fiber optic probes within sealed, temperature- and oxygen-controlled chambers.
Simultaneous Measurements:
- Oxygen concentration and oxygen flux (real-time O₂ consumption rate)
- Mitochondrial and cellular respiration (OXPHOS)
- Reactive oxygen species (ROS) production
- Calcium (Ca²⁺) cycling
- Membrane potential
Sample Types Supported
- Isolated mitochondria
- Permeabilized cells and tissues
- Intact living cells
- Minced or homogenized tissue
- CNS Tissue slices (in near future)
The O2k system provides precise calibration, high sensitivity, and strong signal fidelity, allowing accurate mitochondrial functional assessment in small tissue samples without requiring direct mitochondrial isolation.
Oroboros O2k High-Resolution Respirometry (HRR) System
The Oroboros O2k-Oxygraph provides high-resolution respirometry (HRR) coupled with high-resolution fluorometry for precise mitochondrial functional analysis.
The system utilizes dual Clark-type oxygen electrode chambers and fluorescent fiber optic probes within sealed, temperature- and oxygen-controlled chambers.
Simultaneous Measurements:
- Oxygen concentration and oxygen flux (real-time O₂ consumption rate)
- Mitochondrial and cellular respiration (OXPHOS)
- Reactive oxygen species (ROS) production
- Calcium (Ca²⁺) cycling
- Membrane potential
Sample Types Supported
- Isolated mitochondria
- Permeabilized cells and tissues
- Intact living cells
- Minced or homogenized tissue
- CNS Tissue slices (in near future)
The O2k system provides precise calibration, high sensitivity, and strong signal fidelity, allowing accurate mitochondrial functional assessment in small tissue samples without requiring direct mitochondrial isolation.
SUIT (Substrate–Uncoupler–Inhibitor Titration) Protocols
The Core supports advanced SUIT protocols to interrogate oxidative phosphorylation (OXPHOS) and identify specific bottlenecks within the electron transport chain.
Applications include:
- Mitochondrial physiology studies
- Disease model characterization
- Drug and toxin evaluation
- Functional mitochondrial phenotyping
- Intervention studies
Together, these integrated platforms provide comprehensive, multi-dimensional metabolic phenotyping capabilities for neuroscience, metabolic, inflammatory, and translational research applications.
SUIT (Substrate–Uncoupler–Inhibitor Titration) Protocols
The Core supports advanced SUIT protocols to interrogate oxidative phosphorylation (OXPHOS) and identify specific bottlenecks within the electron transport chain.
Applications include:
- Mitochondrial physiology studies
- Disease model characterization
- Drug and toxin evaluation
- Functional mitochondrial phenotyping
- Intervention studies
Together, these integrated platforms provide comprehensive, multi-dimensional metabolic phenotyping capabilities for neuroscience, metabolic, inflammatory, and translational research applications.
Contact Information
For assistance with Bioenergetics Assays, experimental design, sample preparation, data acquisition, or project scheduling, please contact:
Dr. Samir Patel
COBRE Core Co-Director
skpate2@uky.edu
Dr. Prashant Kunjadiya
COBRE Core Manager – Bioenergetics
pdku223@uky.edu