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Technical Note: Smart Lipid Droplet Assay LIVE
In this Technical Note, we introduce the Smart Lipid Droplet Assay (SLDA) and showcase its output in multiple cell types at varying confluences and across different field of view sizes. We then compare the performance of the SLDA against the fluorescence marker LipidSpot 610, before finishing with a case study where we use the SLDA to quantify how oleic acid (OA) addition changes LD dynamics in pre-adipocyte cells.
Technical Note: Increase biological relevance and simplify your discovery workflow: a case study on cytotoxicity
In this document, we demonstrate how the CX-A can be used in drug screening. We go through the whole process from image acquisition to data analysis and interpretation. As a case study, we chose to test what extent the art of observing influences the outcome of our experiments; something too often ignored when running live cell experiments. We measured the effects on cell health of four different cell treatments (1) label-free imaging and vehicle; (2) label-free imaging and addition of Mitotracker; (3) fluorescence imaging at low intensity regime and addition of Mitotracker; (4) fluorescence imaging at high intensity regime and addition of Mitotracker, using EA.
Technical Note: Live T Cell Assay
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.
Technical Note: EVE Analytics
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.
Feature Application: Calculating kinetic EC50 values from dose-response curves
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.
Feature Application: Characterization of single cells at the population level
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.
Feature Application: Characterization of neurons
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).
Feature Application: Live T Cell Assay
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.
Feature Application: Multiparametric cytotoxicity assays
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.
Occurrence of lymphangiogenesis in peripheral nerve autografts contrasts Schwann cell-induced apoptosis of lymphatic endothelial cells in vitro
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
Published in: Biomolecules
Tags: peripheral nerve regeneration, lymphangiogenesis, Schwann cells, lymphatic endothelial cells