Live Staining of Functional Mitochondria Using TMRM



March 7, 2019

EP145 v1.0

Goals:

Key Steps:

Other Required Materials:

Visualization of functional mitochondria using tetramethylrhodamine, methyl ester (TMRM) reagent via fluorescent imaging in live Organ-Chip

  • Live staining and fluorescent imaging in Organ-Chips
  • Image-iT™ TMRM Reagent (Thermo ScientificTM I34361)
  • Cell culture media (serum-free)

Background

Tetramethylrhodamine, methyl ester (TMRM) is a cell-permeant dye that accumulates in healthy mitochondria with intact membrane potentials.

Method

Sample type

Live Organ-Chip

See Protocol EP155 Live Staining of Cells.

Recommended reagent dilution and incubation time

Dilute 1:1000 (final concentration 0.1 µM to 0.15 µM) in serum-free medium. Incubate for 30 minutes at 37C.

Representative image

Image-iT™ TMRM reagent staining (red) of the active mitochondria and nuclei (blue) in human hepatocytes in the Liver-Chip.

More information on vendor site

https://www.thermofisher.com/order/catalog/product/N1142

Organs-Chips in which the staining has been validated

Image-iT™ TMRM reagent has been validated in both Liver-Chips and Caco-2 Intestine-Chips.


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