ISHE
Holographic microscopy-based method for label-free monitoring of cellular electrogenetransfection – new perspectives for DNA
vaccination
Project Details
Project code: PN-IV-PCB-RO-MD-2024-0285
Contract no.: 62PCBROMD/2025
Project director: Prof. Dr. Mihaela G. Moisescu
Project type: Complex projects – Bilateral with Republic of Moldova
Founding from: Unitatea Executivă pentru Finanțarea Invățământului Superior, a Cercetării, Dezvoltării și Inovării (UEFISCDI)
Status: Ongoing
Starting date: 01.09.2025
Ending date: 31.08.2027
Consortium:
– Carol Davila University of Medicine and Pharmacy, Faculty of Medicine, Biophysics and Cellular Biotechnology Dept.
– Moldova State University, Institute of Applied Physics, Materials for Photovoltaic and Photonics Laboratory
Project Scope
ISHE is a research project intending to upgrade cellular technologies through digital holographic microscopy (DHM) and machine learning models.
The project consists in developing an intelligent method based on digital holographic microscopy imaging for quantifying the electrotransfection efficiency of living cultured cells, in their native culture environment, without using any kind of label. The project is addressed to the domain of cellular bio-technologies and applies techniques from the domain of bio-photonics.
By setting new procedures for evaluating gene delivery, our work advances cell and gene therapies, and improves biotechnological applications of genetically modified cells for medicine
and industry.
The project has an important collaborative impact: the project bridges Romanian and Moldovan scientific expertise, fostering knowledge transfer and research infrastructure. Through our collaboration, we aim to accelerate scientific discovery, expand the practical applications of holographic microscopy in biomedical research and improve the integration of Moldavian researcher into the Romanian scientific community.


What We Do
Our interdisciplinary project focuses on electrotransfection (also known as electro-gene transfer), a technique where electrical pulses are used to deliver genetic material into cells. While electrotransfection is increasingly important in research and medicine due to its efficiency and safety, evaluating its success at the single-cell level has remained challenging—until now.
Using DHM in both classical and polarization-sensitive modes, we non-invasively capture detailed phase images of living cells after gene transfer. These images allow us to extract key biophysical parameters—such as cell dry mass, refractive index, and birefringence—which may correlate with protein content and successful genetic modification.
We further validate and optimize our measurements by combining traditional fluorescence techniques and integrating machine learning algorithms that intelligently analyze the holographic data.
Our technology offers major advantages:
– Label-free analysis: no additional reagents, dyes, or genetic markers needed
– Single-cell accuracy: real-time monitoring of individual living cells
– Minimal sample preparation: preservation of cell viability for extended studies
– High speed and potential for automation: large-scale or high-throughput experiments are facilitated
– Versatility: applicable to a wide range of cell types and transfection techniques
Expected Outcomes
– Development of a polarization-sensitive digital holographic microscopy (DHM) setup
– Automated, single-cell analysis of gene transfer using advanced image processing and machine learning
– Validated correlation between holographic phase parameters and gene expression levels
– Optimized protocols for electrotransfection and cell monitoring
– Publication of joint research in leading scientific journals
– Knowledge transfer and infrastructure development between Romanian and Moldovan research teams
– Enhanced capacity for real-time, non-invasive cell monitoring applicable in biotechnology and clinical research