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Yashraj Biotechnology leverages the ImageXpress Nano to substantially improve their multi-parametric imaging and analyses

Yashraj Biotechnology Limited

Dr. Shweta Bhatt, Chief Scientific Officer and R&D Head

ImageXpress® Nano Automated Imaging System

SpectraMax® iD5 Multi-Mode Microplate Reader

MetaXpress® High-Content Image Acquisition and Analysis Software

The Challenge

Unexpected toxicity in areas such as cardiotoxicity, hepatotoxicity, and neurotoxicity is a serious complication of clinical therapy and one of the key causes for failure of promising drug candidates in development. Animal studies have been widely used for toxicology research to provide preclinical security evaluation of various therapeutic agents under development. Species differences in drug penetration of the blood-brain barrier, drug metabolism, and related toxicity contribute to failure of drug trials from animal models to human. The existing system for drug discovery has relied on immortalized cell lines, animal models of human disease, and clinical trials in humans. Moreover, drug candidates that are passed as being safe in the preclinical stage often show toxic effects during the clinical stage. Only about 16% of drugs, on average, are approved for human use.

Before switching to the ImageXpress Nano, Yashraj Biotechnology used a phase contrast and fluorescence microscope, but found they lacked the ability to take z-stacks of spheroids/organoid models and didn’t have any image analysis software.

Yashraj Biotechnology Team with ImageXpress Nano

“The ImageXpress Nano Automated Imaging System offers a good amount of flexibility in imaging a wide variety of samples ranging from 3D cell aggregates to whole cell and sub-cellular organelles.”
- Dr. Shweta Bhatt, Chief Scientific Officer, Yashraj Biotechnology

The solution

Yashraj Biotechnology is developing cutting-edge research in the stem cell domain and the ImageXpress Nano Automated Imaging System has enabled them to better image their in-house-developed iPSC-derived hepatocytes, cardiomyocytes, and neural cells, as well as several different types of multiparametric assays.

The Results

“The ImageXpress Nano Automated Imaging System offers a good amount of flexibility in imaging a wide variety of samples ranging from 3D cell aggregates to whole cell and sub-cellular organelles. Contrary to conventional fluorescence microscopes, the [ImageXpress Nano] allows focusing across different planes of a 3D structure (spheroid/ organoids). It also allows environmental control over the samples using preset humidity, CO2, and temperature control for time lapse experiments and live cell imaging features. The MetaXpress® High-Content Image Acquisition and Analysis Software enables imaging and analyses using predefined protocols for specific types of experiments while having the flexibility to set custom protocols. Overall, the ImageXpress Nano system is a great platform for multi-parametric and high throughput imaging and analyses.” - Dr. Shweta Bhatt, Chief Scientific Officer, Yashraj Biotechnology

neurite outgrowth assay

Figure: Neurite outgrowth assay: Representative images of iPSCs-derived forebrain motor neurons with ß-tubulin (green) stain shown for the untreated and Taxol-treated cells. Forebrain motor neurons treated with compounds for three days and then fixed and stained with anti-ß-tubulin (TUJ-1).

Pluripotent iPS Cells Express Pluripotency Markers - Oct-4,Nanog and SSEA4

Figure: Pluripotent iPS cells express pluripotency markers. (A), Oct-4 (B) Nanog (C) SSEA4. Nuclei counterstained with DAPI (blue).

Cytoskeleton disruption- iPSC-derived Hepatocytes with Actin Stain for Untreated and Latrunculin-treated Cells

Figure: Cytoskeleton disruption: Representative images of iPSC-derived hepatocytes with actin (green) stain shown for the untreated and Latrunculin-treated cells. Hepatocytes treated with compounds for three days and then fixed and stained with anti-Phalloidin-Actin.

iPSC-derived Cardiomyocytes with TUNEL and Propidium Iodide

Figure: TUNEL assay: Representative images of iPSC-derived cardiomyocytes with TUNEL (green) and Propidium Iodide (red) stain shown for the untreated and Doxorubicin-treated cells. Cardiomyocytes treated with compounds for three days and then fixed and stained with TUNEL and PI.

iPSCs-derived Neural Progenitor Cells Express Neural Progenitor Markers PAX6 and Nestin

Figure: iPSCs-derived neural progenitor cells express neural progenitor markers PAX6 and Nestin. Nuclei counterstained with DAPI (blue).

iPSCs-derived - Tuj1, GFAP and TH with Nuclei Counterstained with DAPI

Figure: A. iPSCs-derived forebrain motor neurons express neural markers Tuj1. B. iPSC-derived astrocytes express neural marker GFAP and C. iPSC-derived midbrain dopaminergic neurons express neural marker thymidine hydroxylase (TH). Nuclei counterstained with DAPI (blue).