Mitochondrial function is measured. Repetitive or continuous measurements are performed of prompt red fluorescence, emerging from the skin due to PpIX build up, and/or delayed fluorescence of PpIX. An estimate of the rate of PpIX generation is used as an indicator of mitochondrial integrity and ATP availability. Mitochondrial oxygen tension is determined from the delayed fluorescence lifetime of PpIX. When blood supply to the measurement volume is interrupted or reduced, the resulting changes to the mitochondrial oxygen tension allow an estimation of information about the kinetics of oxygen consumption in the mitochondria, such as the maximum rate of oxygen consumption as well as the Michaelis-Menten constant, providing information about the oxygen affinity of the mitochondrial respiratory chain.

The present invention relates to an electrode for a biosensor for NADH measurement and a manufacturing method therefor. An electrode manufactured by the method according to the present invention enjoys the advantages of stabilizing current flow during an electric polymerization reaction, making the contact angle of the modified material remarkably small to increase the efficiency of surface modification, and being reusable several times. In addition, when applied to a biosensor for NADH measurement, the electrode of the present invention maintains sensitivity and selectivity at a high level without interference and thus easily measures a target of interest even in blood or serum that necessarily requires a pretreatment process due to the existence of a trace amount of a material to be measured. In addition, when applied to a biosensor for NADH measurement, the electrode can measure cell viability in a continuous manner and in real time, which leads to the application thereof to the cell toxicity assay field, and enables the measurement of cell viability in apoptotic cells lacking the mitochondrial function.

Methods and corresponding apparatus and systems for assessing cellular metabolic activity are disclosed. In one aspect, a cell can be illuminated with optical radiation in order to cause multi-photon excitation of at least one endogenous metabolic cofactor in that cell and cause the excited metabolic cofactor to emit fluorescent radiation. A detector can be used to detect the fluorescent radiation emitted by the excited endogenous metabolic cofactor. A computer processor can analyze the fluorescent radiation to derive the following parameters: (1) using a computer processor to analyze the intensity of the fluorescent radiation, (2) a fluorescence lifetime of at least one of the excited metabolic cofactor, (3) a parameter indicative of mitochondrial clustering in said cell. These parameters can be used to assess at least one metabolic process of the cell.

A method of pretreating a blood sample for measuring ATP of a pathogenic microorganism in blood, comprising: preparing a pellet of platelets and the pathogenic microorganism from the blood sample; and subjecting the pellet of platelets and the pathogenic microorganism to the following steps (A) to (C) in any order (including multiple simultaneous steps). A method of pretreating a cultured blood sample, wherein the method is a simplified method, does not comprise the following step (A), and comprises the steps (B) and (C) only simultaneously performed. (A) digesting cell membrane proteins of the platelets with a protease; (B) swelling the platelets in a hypotonic solution; and (C) disrupting cell membranes of the platelets with a detergent solution under a condition of suppressing an effect on the pathogenic microorganism.

An apparatus (1000) and kit (1000) for the detection of ATP in a liquid sample is provided. The apparatus and kit comprise a liquid reagent composition comprising luciferin and a sampling device having (100) a sampling portion (30) and a handling portion (20). The sampling portion (30) is adapted to acquire and releasably retain a predetermined volume of a liquid sample in one or more cavity (32) that is not substantially defined by space between a plurality of fibers. The sampling device (30) comprises a dry coating that includes an effective amount of a pH-adjusting reagent that, when contacted with a liquid reagent composition having a pH of about 6.8 or lower, changes the pH of the liquid reagent composition to 6.9 or higher. A method of use of the apparatus or kit is also provided.

The present invention relates in general to cellular analysis tools and more particularly to methods and systems for detecting or determining cyclic nucleotide concentrations and related pathway activation in samples. Samples containing cyclic nucleotides may be contacted with a cyclic nucleotide-dependent protein kinase (e.g., cAMP- and cGMP-dependent protein kinases) and a detection system that includes a substrate capable of being phosphorylated by the cyclic nucleotide-dependent protein kinase. The activity of cyclic nucleotide related cell signaling pathways may be measured based on detecting the activity of the cyclic nucleotide-dependent protein kinase.

Methods for identifying animals that are genetically superior, drugs, nutritional strategies, or physiological manipulations that improve feed efficiency or productivity of animals, e.g., selecting animals that are genetically superior for feed efficiency or productivity based on metabolic rates of particular tissues, wherein metabolic rates of certain tissues such as skeletal muscle are inversely proportional to feed efficiency, while metabolic rates of other tissues such as mammary gland are directly proportional to milk production. Thus, animals with low skeletal muscle metabolic rates are generally more feed efficient, e.g., gain more weight per unit of food. The methods herein may be used to improve the genetics, nutrition, and handling or animals more efficiently produced animal products, e.g., meat production, milk, production, egg production, wool production, etc. The methods herein may also be used to determine estimated breeding values of animals for feed efficiency, growth, or production.

Various embodiments of a light detection device and a method of using the device are disclosed. In one or more embodiments, the light detection device can include a housing that extends along a housing axis between top and bottom surfaces. The device can also include a port that is adapted to receive a sample, and a door connected to the housing. The door can include an actuator portion adapted to selectively move the door between a closed position and an open position, and a cover portion connected to the actuator portion and adapted to close the port when the door is in the closed position and open the port when the door is in the open position to allow external access to the port.

The invention relates to the detection of the cofactor reduced nicotinamide adenine dinucleotide phosphate (NADPH). Provided is a sensor molecule for the resonance energy transfer (RET)-based detection of NADPH, the sensor comprising a segment A connected via a linker to a segment B, wherein each of segment A and segment B comprises a member of a RET pair comprising a donor moiety and an acceptor moiety, further characterized in that (i) segment A comprises a binding protein (BP) for NADPH, the BP being dihydrofolate reductase (DHFR; EC 1.5.1.3) or a functional homolog, fragment, derivative or variant thereof, showing the desired NADPH binding properties, and wherein the BP comprises a heterologous protein domain inserted at or replacing at least part of the region corresponding to positions (20) to (27) of E. coli DHFR, said heterologous protein domain comprising the member of the RET pair; (ii) segment B comprises a ligand (L) capable of intramolecular binding to said BP only in the presence of NADPH; such that the donor moiety and the acceptor moiety are in a suitable juxtaposition to yield a RET signal when L is bound to BP, and wherein NADPH-induced binding of L to BP results in an increase in RET efficiency.

When bacteria growth is determined in the related art using an ATP method, the bacteria growth is determined, based on an increase or a decrease in live bacteria ATP with the lapse of a culture time. Accordingly, it is necessary to measure the live bacteria ATP multiple times while antimicrobial susceptibility culture is carried out. It takes time and labor in sample preparation for each measurement, and it is difficult to quickly obtain an antimicrobial susceptibility result. Therefore, according to the present invention, the presence or absence of antimicrobial susceptibility of bacteria is determined, based on an ATP luminescence amount derived from dead bacteria in a culture liquid.