Nexcelom Bioscience

978-327-5340

Detect and Monitor Viability of 3D Tumor Spheroids

  1. Directly image tumor spheroids in various microwell formats
  2. Non-invasive bright field imaging allows the user to image the same plate over multiple days
  3. Perform a two-color fluorescent viability assay

Introduction

The Celigo imaging cytometer has been developed to fully automate imaging and analysis of tumorspheres. This automated morphometric analysis tool significantly reduces the time and effort needed to quantify key aspects of 3D spheres including size, growth, growth tracking over time and response to chemotherapeutics.

Celigo imaging process

Monitor Viability of 3D Tumor Spheroids

Experimental Setup

3D tumor spheroid fluorescence viability assay setup

3D cultures were treated with multiple drugs (17-AAG, Paclitaxel, TMZ or Doxorubicin) and stained for viability after 21 days.

Live / dead stains used were calcein AM (green), Propidium Iodide (red) to measure live and dead cells on days 4, 7, 10, 14, and 17. Plates containing spheroids were imaged on Celigo imaging cytometer.

  • 17-AAG decreased sphere size and caused the most significant cell death
  • Paclitaxel decreased sphere size but maintained a significant number of live cells
  • Temozolomide caused no significant decrease in sphere size and did not cause cell death
  • Doxorubicin decreased sphere size and caused significant cell death
 
3D tumor spheroids

Multi-drug Viability 3D Assay

multidrug viability assay

Spheroid diameter (black) and total live fluorescent intensity (green) after drug treatment.


3D Tumor Spheroids Treated at Different Drug Concentrations and Stained with Propidium Iodide

Control

3D tumor spheroid control

Medium Drug Dose

3D tumor spheroid medium drug dose

High Drug Dose

3D tumor spheroid high drug dose

Measuring Viability 3D Spheroids with Morphological Differences

3D tumor spheroid control
3D spheroids with morphological differences
3D tumor spheroid medium drug dose
3D spheroids with morphological differences
 
3D tumor spheroid control
3D spheroids with morphological differences
3D tumor spheroid medium drug dose
3D spheroids with morphological differences
 
3D tumor spheroid control
3D spheroids with morphological differences
3D tumor spheroid medium drug dose
3D spheroids with morphological differences
 
3D tumor spheroid control
3D spheroids with morphological differences
3D tumor spheroid medium drug dose
3D spheroids with morphological differences

3D Tumor Spheroids Treated with Various Compounds and Stained with Calcein-AM and PI

Treated 3D Tumor Spheroids and Stained with Calcein-AM and PI
Treated 3D Tumor Spheroids and Stained with Calcein-AM and PI
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