Scientific Publications

This technical note explores these challenges, and presents Nanolive’s cutting edge, AI-assisted solution, the Live T Cell Assay. It also includes a full use case study where the Live T Cell Assay is used to quantify the effect a bispecific antibody has on T cell and cancer cell dynamics.

Nanolive’s label-free technology makes it possible to image cells for long periods of time, at high temporal resolution. The quantity and complexity of the images generated allows us to visualize biological processes in unprecedented detail, but also magnifies the challenges associated with image analysis. Manual image registration and analysis is impossible and so computer-aided processing must be used to harness data complexity. In this technical note, we introduce the key elements involved in cell segmentation, which are essential to understand the novelty of EVE Analytics (EA), Nanolive’s software solution for quantitative cell analysis. We then evaluate the performance of EA segmentation against fluorescence-based segmentation and compare how metrics produced by both approaches differ.

The non-invasive nature of label-free imaging means cells can be continuously monitored, over infinite periods of time, which means kinetic EC50 values can also be calculated. Time-dependent EC50 values provide information about drug stability; whether a drug’s potency increases or decreases over time. In this application note we show how to calculate kinetic EC50 values from data directly output from Nanolive’s image analysis software, EVE Analytics.

In this Feature Application, we showcase the enormous potential that Nanolive live cell imaging holds for single cell characterization. We begin, by analyzing micro-heterogeneity at the population and the temporal level in unperturbed cells (case study one). We then extend our analysis to include a quantitative assessment of lipid droplet dynamics (case study two), before investigating how intracellular trafficking (case study three) and respiratory perturbation (case study four) impacts microheterogeneity in cell morphology.

This Feature Application shows the huge potential that Nanolive cell imaging holds for neurobiological research. Our first case study shows a timeline of the morphological changes undifferentiated primary cortical neurons undergo after exposure to neurite stimulation media. High precision segmentations are used to calculate cell metrics (e.g. volume, shape and dry mass) of interest. These calculations are directly linked to novel behaviours observed in individual neurons. Our second and third case studies focus on the sub-cellular morphological changes that stem cells undergo during neural differentiation, at both the single cell (case study 2) and population level (case study 3).

The Live T Cell Assay promises to accelerate the advancement of basic cell biology and of novel, safe, and effective immunotherapies. The goal of this application note is to show some of the potential uses of the Live T Cell Assay and hopefully, inspire research that will increase the success rate of anticancer drug development. To this end, we present the first in-depth investigation of bispecific antibodies using the Live T Cell Assay.

This Feature Application shows the huge potential that Nanolive imaging holds for the drug discovery process, where cytotoxicity remains one of the major causes of drug withdrawal, and there is an urgent need for reliable and time-saving assay workflows. Our first case study features an in-depth investigation of the dynamics of cell death following exposure to seven cell death stimuli: extreme shifts in external pH, thermal stress, phototoxicity, oxidative stress, Shikonin, Ebastine, and Taxol. Our second case study shows that we can quantify drug-dependent and dose-dependent effects in 96-well plate format.

Authors: Carina Hromada, Jaana Hartmann, Johannes Oesterreicher, Anton Stoiber, Anna Daerr, Barbara Schädl, Eleni Priglinger, Andreas H. Teuschl-Woller, Wolfgang Holnthoner, Johannes Heinzel, David Hercher

https://doi.org/10.3390/biom12060820, 2022

Published in: Biomolecules

Tags: peripheral nerve regeneration, lymphangiogenesis, Schwann cells, lymphatic endothelial cells

Authors: Penghao Ji, Shenyan Zhang, Peilei Liu, Xi Li, Weichao Bao, Xiangzhi Cui, Minfeng Huo, Jianlin Shi

https://doi.org/10.1016/j.nantod.2022.101511, 2022

Published in: Nanotoday

Tags: Pyroptosis; Oligomycin A; Mitochondrial damage; Oxidative stress; Mild hyperthermia

Authors: Shanshan Zheng, Yue Cai, Yulu Hong, Yubei Gong, Licheng Gao, Qingyong Li, Le Li, Xuanrong Sun

https://doi.org/10.1080/10717544.2022.2081380, 2022

Published in: Drug Delivery

Tags: Peptide–drug conjugate; legumain-responsive; pH-responsive; synergistic cancer therapy