Nexcelom Bioscience


Image and measure the size of 3D tumor spheroids under multiple drug and growth conditions over time

  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. Measure spheroid volume and diameter concurrently with spheroid imaging


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

Drug-Induced Tumor Spheroid Growth Inhibition

Experimental Setup

tumor spheroid growth setup

4-day old spheroids were treated with compounds for 72 hours. Drug-induced concentration-dependent growth inhibition was obtained.

  • Day 0-4: form spheroids
  • Day 4-7: drug treatment
  • Day 4-14: measure spheroid size

Drug Concentration

drug concentration

Green calcein AM stained target cells

U 87-MG (giloblastoma) + 17-AGG

U 87-MG (giloblastoma) + 17-AGG

Green calcein AM stained target cells

Measuring Spheroid Growth Inhibition under Drug Treatment and Hypoxia

Experimental Protocol:

  1. Seed 2500 NCI-H460 cells/well in a ULA 96-well plates
  2. On day 4 treat with drug X or vehicle control
  3. Place one plate in hypoxic conditions and one plate at normoxic condition
  4. On day 4 post treatment image entire 96-well plate in ~5 minutes

At high drug dose, NCI-H460 3D tumor spheroids showed growth inhibition under hypoxic conditions

NCI-H460 Spheroid at Normoxia, treated with
Drug X at 10 µM

Spheroid at Normoxia, treated with Drug X

NCI-H460 cells treated with Drug X and grown in Normoxia vs Hypoxia conditions

high drug dose 3D tumor spheroid growth inhibition

NCI-H460 Spheroid at Hypoxia, treated with
Drug X at 10 µM

Spheroid at Hypoxia, treated with Drug X

Captured bright field images of NCI-H460 spheroids treated with a 10 µM drug and grown at different oxygen conditions show that spheroid size is affected by growth conditions.

By imaging the same plate over multiple days, drug-treated spheroids at hypoxic conditions had a smaller spheroid diameters compared to the spheroids grown at normal conditions.