Live Staining of Mitochondria Using MitoTracker® Fluorescent Dyes



EP144 v1.0

Goals:

Key Steps:

Other Required Materials:

Visualize mitochondria in live cells via fluorescent imaging in Organ-Chips

  • Live staining and fluorescent imaging of Organ-Chips
  • MitoTracker® Orange CMTMRos (Thermo ScientificTM M7510), Mitotracker® Red CM-H2XROS (Thermo ScientificTM M7513), or Mitotracker ® Deep Red FM (Thermo Scientific TM M22426)
  • Cell culture medium (serum-free)
  • Dimethyl sulfoxide (DMSO)
  • Fluorescence microscope

Optional

  • PBS
  • 4% paraformaldehyde (PFA)

Introduction

MitoTracker® is a fluorescent dye that stains mitochondria in live cells. Its accumulation is dependent upon membrane potential. This endpoint is used as a marker of cell viability.

Method

Sample type

Live Organ-Chip

See Protocol EP155 Live Staining.

Recommended reagent dilution and incubation time

Prepare 1 mM stock solution in DMSO.

Dilute stock 1:5000 in serum-free media. Incubate for 30 minutes at 37C.

Fixative (optional)

Chips can be fixed immediately after staining: 4% PFA for 15 minutes at room temperature.

The signal remains for up to 24 hours after fixation.

Note: Do not fix cells before MitoTracker® staining.

See Emulate Fixation and Immunofluorescence (IF) Staining for Organ-Chips (#6137).

Representative image

Image of mitochondria (red) and nuclei (blue) in human hepatocytes in the Liver-Chip, stained by the MitoTracker® reagent (top channel).

More information on vendor site

https://www.thermofisher.com/order/catalog/product/M7510?cid=TechComm3a

Organs-Chips in which the staining has been validated

MitoTracker® (Thermo Fisher Scientific TM M7510) has been validated on Liver-Chips.

MitoTracker® (Thermo Fisher Scientific TM M7513 and M22426) have been validated on Caco-2 Intestine-Chips.

 


 

© Emulate, Inc., 2019. All rights reserved.

Emulate® and the Emulate logo are registered trademarks for Emulate, Inc.

MitoTracker® is a registered trademark of Thermo Fisher Scientific.

The technology disclosed in this document may be covered by one or more patents or patent applications owned by or licensed to Emulate, Inc. No license is granted herein. You are solely responsible for determining whether you have all intellectual property rights that are necessary for your intended use of Emulate products or protocols, and whether you are required to obtain any additional intellectual property rights from a third party. Further information is available by contacting Emulate.