Electroporation has been shown to have considerable potential in biological research and clinical applications. This technique permits the opening of reversible or irreversible nanometric pores in cellular membranes by the application of a high-intensity electric field characterized by extremely short duration (i.e., microseconds, nanoseconds down to tens of picoseconds). Recently, also thanks to the spreading use of lab-on-chip, it was discovered that applying a high electric field to single cells can cause them to contract and generate blebs. Even though these effects are known in the literature, their correlation with the electroporation phenomenon is still unclear. For this reason, in this work, we present a reliable, low-cost sensing system based on a lab-on-chip platform with transferred laser-induced graphene (LIG) electrodes to study electroporation by bright-field and fluorescence time-lapse microscopy. The acquired video was then used to extrapolate the calcium intake due to electroporation, cell contractility, and morphology over time. The two abovementioned side effects were finally correlated with electroporation and used to estimate its efficacy on single cells, exploiting calcium intake information as the ground truth. The proposed sensing system could be used to predict the electroporation phenomenon easily and even in a label-free modality, using exclusively the morphological variations induced in the single cells.

Lab-on-Chip Label-Free Sensing System for Electroporation Based on Time-Lapse Microscopy

Casciati A.;Tanori M.;Zambotti A.;Merla C.;
2024-01-01

Abstract

Electroporation has been shown to have considerable potential in biological research and clinical applications. This technique permits the opening of reversible or irreversible nanometric pores in cellular membranes by the application of a high-intensity electric field characterized by extremely short duration (i.e., microseconds, nanoseconds down to tens of picoseconds). Recently, also thanks to the spreading use of lab-on-chip, it was discovered that applying a high electric field to single cells can cause them to contract and generate blebs. Even though these effects are known in the literature, their correlation with the electroporation phenomenon is still unclear. For this reason, in this work, we present a reliable, low-cost sensing system based on a lab-on-chip platform with transferred laser-induced graphene (LIG) electrodes to study electroporation by bright-field and fluorescence time-lapse microscopy. The acquired video was then used to extrapolate the calcium intake due to electroporation, cell contractility, and morphology over time. The two abovementioned side effects were finally correlated with electroporation and used to estimate its efficacy on single cells, exploiting calcium intake information as the ground truth. The proposed sensing system could be used to predict the electroporation phenomenon easily and even in a label-free modality, using exclusively the morphological variations induced in the single cells.
2024
Electroporation
image analysis
lab-on-a-chip
label free
laser-induced graphene (LIG)
machine learning
time-lapse microscopy
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12079/84328
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