In recent years, intracellular LDs have been discovered to play an important role in several pathologies. Therefore, detection of LDs would provide an in-demand diagnostic tool if coupled with flow-cytometry to give significant statistical analysis and especially if the diagnosis is made in full non-invasive mode. Here we combine the experimental results of in-flow tomographic phase microscopy with a suited numerical simulation to demonstrate that intracellular LDs can be easily detected through a label-free approach based on the direct analysis of the 2D quantitative phase maps recorded by a holographic flow cytometer. In fact, we demonstrate that the presence of LDs affects the optical focusing lensing features of the embracing cell, which can be considered a biological lens. The research was conducted on white blood cells (i.e., lymphocytes and monocytes) and ovarian cancer cells. Results show that the biolens properties of cells can be a rapid biomarker that aids in boosting the diagnosis of LDs-related pathologies by means of the holographic flow-cytometry assay for fast, non-destructive, and high-throughput screening of statistically significant number of cells.

Inhibition of respiratory complex I (CI) is becoming a promising anti-cancer strategy, encouraging the design and the use of inhibitors, whose mechanism of action, efficacy and specificity remain elusive. As CI is a central player of cellular bioenergetics, a finely tuned dosing of targeting drugs is required to avoid side effects. We compared the specificity and mode of action of CI inhibitors metformin, BAY 87-2243 and EVP 4593 using cancer cell models devoid of CI. Here we show that both BAY 87-2243 and EVP 4593 were selective, while the antiproliferative effects of metformin were considerably independent from CI inhibition. Molecular docking predictions indicated that the high efficiency of BAY 87-2243 and EVP 4593 may derive from the tight network of bonds in the quinone binding pocket, although in different sites. Most of the amino acids involved in such interactions are conserved across species and only rarely found mutated in human. Our data make a case for caution when referring to metformin as a CI-targeting compound, and highlight the need for dosage optimization and careful evaluation of molecular interactions between inhibitors and the holoenzyme.

Anticancer strategies aimed at inhibiting Complex I of the mitochondrial respiratory chain are increasingly being attempted in solid tumors, as functional oxidative phosphorylation is vital for cancer cells. Using ovarian cancer as a model, we show that a compensatory response to an energy crisis induced by Complex I genetic ablation or pharmacological inhibition is an increase in the mitochondrial biogenesis master regulator PGC1α, a pleiotropic coactivator of transcription regulating diverse biological processes within the cell. We associate this compensatory response to the increase in PGC1α target gene expression, setting the basis for the comprehension of the molecular pathways triggered by Complex I inhibition that may need attention as drawbacks before these approaches are implemented in ovarian cancer care.

Live cells act as biological lenses and can be employed as real‐world optical components in bio‐hybrid systems. Imaging at nanoscale, optical tweezers, lithography and also photonic waveguiding are some of the already proven functionalities, boosted by the advantage that cells are fully biocompatible for intra‐body applications. So far, various cell types have been studied for this purpose, such as red blood cells, bacterial cells, stem cells and yeast cells. White Blood Cells (WBCs) play a very important role in the regulation of the human body activities and are usually monitored for assessing its health. WBCs can be considered bio‐lenses but, to the best of our knowledge, characterization of their optical properties have not been investigated yet. Here, we report for the first time an accurate study of two model classes of WBCs (i.e., monocytes and lymphocytes) by means of a digital holographic microscope coupled with a microfluidic system, assuming WBCs bio‐lens characteristics. Thus, quantitative phase maps for many WBCs have been retrieved in flow‐cytometry (FC) by achieving a significant statistical analysis to prove the enhancement in differentiation among sphere‐like bio‐lenses according to their sizes (i.e., diameter d) exploiting intensity parameters of the modulated light in proximity of the cell optical axis. We show that the measure of the low intensity area (S: Iz

In recent years, the dynamic role of lipid droplets (LDs) in many cellular activities has been increasingly brought to light. In fact, it has been discovered that LDs are involved in many pathologies (e.g., diabetes, atherosclerosis, pathogen infections, neurodegenerative diseases and cancer). Moreover, it has been demonstrated that their number and size increase during an inflammation or infectious inside the immune cells, also with the COVID-19. Therefore, detecting LDs within single cells could aid the diagnosis of several pathologies. Currently, the gold-standard technique in this field is Fluorescence Imaging Flow Cytometry (FIFC), in which the single-cell analysis of fluorescence microscopy is implemented in high-throughput modality thanks to the flow-cytometry module. However, to overcome the drawbacks related to the fluorescence staining, Holographic Imaging Flow Cytometry (HIFC) has gaining momentum as label-free alternative to the FIFC tool. Thanks to the interferometric principles at the basis of digital holography, it has been already demonstrated that a suspended cell acts as a biological lens with specific focusing features. Here we show that the presence of intracellular LDs inside the cell is able to change its focalization features, measured through a HIFC system. Therefore, based on this property, we demonstrate that a detection of single cells containing intracellular LDs is possible by means of a direct analysis of the digital holograms recorded in flow cytometry modality. The attained results open the route to the development of a fast, non-destructive, and high-throughput tool for the diagnosis of LDs-related pathologies by exploiting the biolens' signature in HIFC.