We are very proud and excited to announce a new publication from Dr. José Padrón and lead author Adrián Puerta showing the benefits of using Nanolive’s label-free data analysis in Phenotypic Drug Discovery (PPD) programs. Dr. Padrón is a Professor at the Universidad de La Laguna in the Canary Islands (Spain) and the founder of BioLab; a PPD program that aims to identify novel pharmacologically active small-molecules and define their mechanism of action. Dr. Padrón bought a CX-A equipped with EVE Analytics (EA) in August 2021 and wasted no time installing it into his lab’s workflow. This blogpost summarizes the main findings of the paper. The original paper can be downloaded here.
Nanolive’s label-free, multiplexed data offers users a non-invasive means of gathering high content data (11 metrics) without time-consuming experiments or costly fluorescent reagents, which perturb cell responses. In particular, it allows researchers to phenotypically profile small molecules:
- At multiple time-points without biasing results by choosing a single timepoint in end-point techniques
- Without confounding effects of photobleaching or phototoxicity
- In a robust and standardized manner using meaningful metrics of the highest biological relevance, i.e., changes in cell morphology and cell composition
The goal of the study was to evaluate whether Nanolive’s label-free live cell image analysis could be combined with machine learning to identify the mechanism of action of small molecule drugs.
Dr. Padrón and his team tested this by exposing the non-small cell lung cancer cell line SW1573 to three antimitotic drugs: paclitaxel (PTX), colchicine (CLC), and vinblastine (VBL), whose mechanisms of action differ. PTX is a microtubule stabilizer agent; it binds to tubulin, which induces mitotic arrest by preventing the formation of a centered mitotic spindle. In contrast, CLC and VBL are microtubule-disrupting agents; they bind to specific β-tubulin binding sites, which disrupts the formation of microtubules and blocks the cell cycle.
Images were captured every 3 mins for 20 h and then analyzed by Nanolive’s software, EA, which outputs 11 metrics covering all aspects of cell morphology (area, perimeter, form factor, extent, compactness, and eccentricity) and composition (mean RI, average dry mass density, dry mass, and granularity).
The metrics easily captured the phenotypic differences between the two groups of drugs; exposure to PTX caused a reduction in cell area, with a rounded and diminished nucleus area surrounded by a contracted cytoplasm, whereas CLC and VBL induced a nuclear reduction, cytoplasm contraction and an overall reduction in cell area.
When the experiments were completed, the authors used an approach called Fast Fourier Transform (FFT) Analysis to analyze the data. The 11 phenotypic metrics output by EA were fed into an algorithm which broke each parameter down into two measures (maximal amplitude and phase). These values were then used to cluster compounds with similar modes of action with great success.
This study is an important first step towards developing computer-assisted phenotype annotation of the mode of action of compounds. Congratulations to all the authors involved.
Read our latest news
Next upcoming event
Next product launch - 26.06.2023
17 new scientific publications featuring Nanolive technology
Since the start of the year, 17 more papers have been published using Nanolive imaging, featuring an incredibly broad range of cell types; stem cells, adipocytes, neurons, osteoblasts, neutrophils, melanocytes, e. coli, primary synoviocytes, and several cancer cell...
Nanolive imaging confirms success of mesoestetic’s new treatment at the subcellular level
We are happy to see that Nanolive imaging has been used to confirm the success of another new therapy, this time treating melasma. In their latest paper “Depigmenting topical therapy based on a synergistic combination of compounds targeting the key pathways involved...