Alicia Méndez’s research while affiliated with University of La Laguna and other places

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Publications (4)


Electrochemical Determination of Serotonin Exocytosis in Human Platelets with BDD-on-Quartz Multielectrode Array Biosensors
  • Conference Paper
  • Full-text available

May 2024

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9 Reads

Rosalía González-Brito

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Alicia Méndez

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[...]

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Figure 5. Heterogeneity of secretory spikes from human platelets. The figure shows some amperometric spikes obtained from a single experiment. Each trace corresponds to the serotonin release single event exocytosis recorded with BDD-on-quartz MEA devices. Notice the different shapes and kinetics (a-d). All spikes are on the same scale. Notice the different kinetics and quantum sizes, expressed in pC. Spike characterization was carried out using our macros for IgorPro 8 ® [35]. These macros can be freely shared upon request.
Comparison of the structural materials adopted for the three MEA technologies.
Analytical Determination of Serotonin Exocytosis in Human Platelets with BDD-on-Quartz MEA Devices

January 2024

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46 Reads

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1 Citation

Biosensors

Amperometry is arguably the most widely used technique for studying the exocytosis of biological amines. However, the scarcity of human tissues, particularly in the context of neurological diseases, poses a challenge for exocytosis research. Human platelets, which accumulate 90% of blood serotonin, release it through exocytosis. Nevertheless, single-cell amperometry with encapsulated carbon fibers is impractical due to the small size of platelets and the limited number of secretory granules on each platelet. The recent technological improvements in amperometric multi-electrode array (MEA) devices allow simultaneous recordings from several high-performance electrodes. In this paper, we present a comparison of three MEA boron-doped diamond (BDD) devices for studying serotonin exocytosis in human platelets: (i) the BDD-on-glass MEA, (ii) the BDD-on-silicon MEA, and (iii) the BDD on amorphous quartz MEA (BDD-on-quartz MEA). Transparent electrodes offer several advantages for observing living cells, and in the case of platelets, they control activation/aggregation. BDD-on-quartz offers the advantage over previous materials of combining excellent electrochemical properties with transparency for microscopic observation. These devices are opening exciting perspectives for clinical applications.


Figure 1. (A) CV plots recorded at 50 mV/s with 6 different concentrations of adrenaline. (B) Current intensities measured at 0.8 V during the rising ramp, with a linear fit. The electrochemical activity drops slightly at the highest concentration.
Figure 2. Diamond-based MEA device for amperometric recording of platelet exocytosis. (A), General view of the recording system. 1, When using transparent BDD-on-quartz MEAs, the front-end unit can be placed on the stage of an inverted microscope to examine platelet distribution. 2, The USB multi I/O unit and the analog low-pass filter plug-in board. 3, The acquisition program written in LabView ® and a PC complete the system. (B), Faraday cage containing the MEA chip and amplifiers. (C), A BDD-on-quartz MEA showing the 16-electrode layout. (D), Magnified view of the red inset in C. Isolated human platelets on a single electrode observed by DIC microscopy: we, the open BDD spot acting as a working electrode; d, the passivated BDD structure acting as a planar electrical connection. Calibration bar =50 μm. (E), BDD-on-silica MEA showing the gold contact pads. These opaque MEAs have the same 16-electrode layout as in panel C. (F), a 10 s recording from the 16 auto-scalable traces show three amperometric spikes in channels 6, 9, and 10.
Multielectrode Arrays as a Means to Study Exocytosis in Human Platelets

January 2023

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79 Reads

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7 Citations

Biosensors

Platelets are probably the most accessible human cells to study exocytosis by amperometry. These cell fragments accumulate biological amines, serotonin in particular, using similar if not the same mechanisms as those employed by sympathetic, serotoninergic, and histaminergic neurons. Thus, platelets have been widely recognized as a model system to study certain neurological and psychiatric diseases. Platelets release serotonin by exocytosis, a process that entails the fusion of a secretory vesicle to the plasma membrane and that can be monitored directly by classic single cell amperometry using carbon fiber electrodes. However, this is a tedious technique because any given platelet releases only 4–8 secretory d-granules. Here, we introduce and validate a diamond-based multielectrode array (MEA) device for the high-throughput study of exocytosis by human platelets. This is probably the first reported study of human tissue using an MEA, demonstrating that they are very interesting laboratory tools to assess alterations to exocytosis in neuropsychiatric diseases. Moreover, these devices constitute a valuable platform for the rapid testing of novel drugs that act on secretory pathways in human tissues.

Citations (2)


... We detected amperometric recordings from the quantum release of serotonin from human platelets with boron-doped nanocrystalline diamond on MEA systems (BDD-MEA) of 16 microelectrodes. Our studies were carried out with devices on silicone matrices (BDD-on-silicon MEA) [35] and quartz substrates (BDD-on-quartz MEA) [36]. ...

Reference:

Abstract of the 4th International Online Conference on Crystals
Analytical Determination of Serotonin Exocytosis in Human Platelets with BDD-on-Quartz MEA Devices

Biosensors

... The atoms' connectivity profiles were processed as described by Michos and Raptis [33] to derive Chemical Equitable Partitions. Electrostatic lattice site potentials were calculated using VESTA's [34] built-in functionality. CEP cells were compared to the atom groups defined by Wyckoff positions, on the one hand, and site potentials, on the other. ...

Multielectrode Arrays as a Means to Study Exocytosis in Human Platelets

Biosensors