Project leader: Dr. Aleksandar Krmpot
Project collaborators: Dr. Ivana Drvenica, Dr. Drenka Trivanović, Dr. Mihajlo Radmilović, Dr. Stanko Nikolić, Dr. Danica Pavlović
Duration of the project: 1.9.2020–31.8.2022.
Leading institution: Institute of Physics, University of Belgrade
Collaborating institution: Institute for Medical Research, University of Belgrade
Description of the project:
In this project, we are aiming to utilize state-of-the art optical microscopic techniques expanding their applicability for erythrocytes and erythrocyte derivatives imaging as a potential diagnostic method for modern age disorders, such as cancer, diabetes and other inflammatory/autoimmune diseases. Our research will be mostly based on two-photon excitation fluorescence microscopy (TPEF) and recently demonstrated intrinsic feature of hemoglobin to become a fluorescent molecular target upon two-photon excitation by ultra-short laser pulses. We will unite our “know-how” on TPEF with third harmonic generation (THG) imaging of erythrocytes and complement it with quantitative information obtained by fluorescence correlation spectroscopy (FCS). The latter two will be developed within this project. Using erythrocytes as sensitive markers, we will demonstrate how combining the data acquired by custom–made techniques and established protocols for in vitro and ex vivo hemoglobin based imaging can contribute to a better understanding of the underlying biological basis of specific disorders. This is the first step in development of novel methods for diagnosis and disease risk stratification. We will also boost our initiated research on TPEF microscopy as an effective tool for label- and fixation -free imaging of erythrocytes and their membranes at single cell level. The research revealed the relation between spatial distribution of hemoglobin within erythrocytes and their morphology implying that this relation is an indicator of erythrocytes functional status.
Six researchers from two leading national institutions and from various, complementary, fields (physics, biology, biophotonics, pharmacy) will carry out the activities divided into 7 tasks and deliver 13 items, amongst two new techniques will be developed: THG and FCS for the benefit to the broad international and Serbian scientific community. HEMMAGINERO is supported by DESY, Germany and Karolinska Institutet, Sweden formally and by open access to the equipment. The project will be realized in 2 years and by cca 200,000EUR employing and upgrading the most valuable equipment in Serbian scientific institutions and respecting all the safety and ethical issues.