1 essenbioscience.com/IncuCyte

Assay Concept:

• Measure cell proliferation either label-free or fluorescently labeled using live cell time-lapse imaging. Easily generate long-term growth and growth-inhibition curves as well as monitor morphology.

• Directly measure changes in true cell number (nuclear count) over time using IncuCyte™ NucLight™ live cell labeling reagents. Calculate growth rates and doubling times and explore co-cultures.

• Multiplex proliferation readouts with measurements of apoptosis or cytotoxicity by combining with the IncuCyte™ Caspase 3/7 Reagent or IncuCyte™ Cytotox Reagents.

IncuCyte™ Proliferation AssayReal-time quantitative analysis of proliferation

Observe cell proliferation over time and measure proliferation with or without labels in real time.

Automatic analysis of growth curves and doubling times within your incubator.

Simple, flexible 96/384-well protocols - no washing, no fixing, no lifting of cells.

Multiplex with other cell health measurements - differentiate cytostatic and cytotoxic effects.

Identify and quantify cells in co-culture using NucLight™ live cell labeling reagents.

HUVEC were infected with the NucLight™ Red lentivirus reagent, replated, serum starved for 2 hours and treated with basal media containing 1% FBS, and increasing amounts of bFGF (left), or a constant 10ng/ml bFGF and increasing concentrations of suramin (right) . Images were acquired every 3 hours in an IncuCyte® ZOOM Instrument with either a 4x objective (left) or 10x objective (right). Nuclei were counted over the full course of the assay. EC50 and IC50 calculations were made using area under the curve (AUC) values for each condition.

2 essenbioscience.com/IncuCyte8000-0113-D00

IncuCyte™ Proliferation AssayReal-time quantitative analysis of proliferation

Essen BioScience, 300 West Morgan Road, Ann Arbor, Michigan, USA 48108 © 2016 Essen BioScience. All rights reserved. All trademarks are the property of Essen BioScience unless otherwise specified.

Learn more at essenbioscience.com/applications. Or contact us at sales@essenbio.com.

Key applications using the IncuCyte® ZOOM Live-Cell Analysis System

Ordering informationProduct Quantity Cat. No.

IncuCyte™ NucLight™ Red BacMam 3.0 Reagent for nuclear labeling 1 mL 4621

IncuCyte™ NucLight™ Green BacMam 3.0 Reagent for nuclear labeling 1 mL 4622

IncuCyte™ NucLight™ Red Lentivirus Reagent (EF-1 α, Puro) for nuclear labeling 0.2 mL 4625

IncuCyte™ NucLight™ Green Lentivirus Reagent (EF-1 α, Puro) for nuclear labeling 0.2 mL 4624

IncuCyte™ Caspase 3/7 Reagent for apoptosis 20 μL 4440

IncuCyte™ Cytotox Red Reagent for counting dead cells 5 μL x 5 4632

IncuCyte™ Cytotox Green Reagent for counting dead cells 5 μL x 5 4633

HT1080 cells infected with the NucLight™ Red lentivirus reagent were treated with increasing concentrations of cycloheximide (CHX) and imaged at 4x in an IncuCyte® ZOOM Instrument every 3 hours (n=3 wells per treatment in a 96-well plate). Although no alterations in cell morphology were observed, concentration dependent inhibition of cell proliferation was detected. Area under the curve (AUC) values were calculated for each treatment group and used to determine the IC50 value of this compound. Doubling times were calculated using exponential growth equation in GraphPad Prism 5.0. Representative images were taken at 48 hrs.

3D-Spheroids Angiogenesis Apoptosis Cell Culture QC Dilution CloningCytotoxicity Neurite Dynamics - Label-Free

Neuronal Co-Culture - Fluorescence

Phagocytosis Proliferation - Cell Count

Proliferation - Confluence

Reporter Gene Transfection Efficiency

Stem Cell Monitoring & Reprogramming

Immune Cell Killing, Clustering & Proliferation

Scratch Wound Migration & Invasion

ChemotaxisMigration & Invasion