Real-time in vivo imaging of extracellular ATP in the brain with a hybrid-type fluorescent sensor

Adenosine 5’ triphosphate (ATP) is a ubiquitous extracellular signaling messenger. Here, we describe a method for in-vivo imaging of extracellular ATP with high spatiotemporal resolution. We prepared a comprehensive set of cysteine-substitution mutants of ATP-binding protein, Bacillus FoF1-ATP synthase ε subunit, labeled with small-molecule fluorophores at the introduced cysteine residue. Screening revealed that the Cy3-labeled glutamine-105 mutant (Q105C-Cy3; designated ATPOS) shows a large fluorescence change in the presence of ATP, with submicromolar affinity, pH-independence, and high selectivity for ATP over ATP metabolites and other nucleotides. To enable in-vivo validation, we introduced BoNT/C-Hc for binding to neuronal plasma membrane and Alexa Fluor 488 for ratiometric measurement. The resulting ATPOS complex binds to neurons in cerebral cortex of living mice, and clearly visualized a concentrically propagating wave of extracellular ATP release in response to electrical stimulation. ATPOS should be useful to probe the extracellular ATP dynamics of diverse biological processes in vivo.

Design and Implementation Model for Linearization Sensor Characteristic by FPAA

Linearization sensors characteristics becomes very interest field for researchers due to the importance in enhance the system performance, measurement accuracy, system design simplicity (hardware and software), reduce system cost, ..etc. in this paper, two approaches has been introduced in order to linearize the sensor characteristics; first is signal condition circuit based on lock up table (LUT) which this method performed for linearize NTC sensor characteristic. Second is ratiometric measurement equation which this method performed for linearize LVDT sensor characteristic. The proposed methods has been simulated by MATLAB, and then implemented by using Anadigm AN221E04 Field Programmable Analog Array (FPAA) development kit which several experiments performed in order to improve the performance of these approaches.