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Difference between revisions of "TMRM"

From Bioblast
Line 17: Line 17:


:::'''Preparation of 5 mM stock solution (dissolved in DMSO)''':
:::'''Preparation of 5 mM stock solution (dissolved in DMSO)''':
::::# Weigh 2.5 mg of TMRM and dissolve in 1 ml DMSO.
::::# Weight 2.5 mg of TMRM and dissolve in 1 mL DMSO;
::::# Divide into 0.2 ml portions (use dark vials)
::::# Divide into 0.2 mL portions (use dark vials);
::::# Store at -20 °C.
::::# Store at -20 °C.


:::'''Preparation of 1 mM solution - use for experiment''':
:::'''Preparation of 1 mM solution - use for experiment''':
::::# Dilute a 200µl aliquot with 800µl DMSO to reach 1 mL final volume
::::# Dilute a 200 µL aliquot with 800 µL DMSO to reach 1 mL final volume;
::::# Divide into 0.2 ml portions (use dark vials)
::::# Divide into 0.2 mL portions (use dark vials);
::::# Store at -20 °C.
::::# Store at -20 °C.



Revision as of 11:53, 14 August 2018


high-resolution terminology - matching measurements at high-resolution


TMRM

Description

TMRM ( tetramethylrhodamine methyl ester) is an extrinsic fluorophore and can be used as a probe to determine changes in mitochondrial membrane potential. TMRM is a lipophilic cations that is accumulated by mitochondria in proportion to Δψmt. Upon accumulation of the dye it exhibits a red shift in its absorption and fluorescence emission spectrum. The fluorescence intensity is quenched when the dye is accumulated by mitochondria.

Abbreviation: TMRM

Reference: Ehrenberg 1988 Biophys J


MitoPedia methods: Fluorometry 


Application in HRFR

TMRM: Tetramethyrhodamine methyl ester perchlorite (C25H25CIN2O7);
Sigma T 5428 or ThermoFischer T 668, 25 mg, store at -20°C; FW = 500.93


Caution: Light sensitive (store solution in a dark vial)!


Preparation of 5 mM stock solution (dissolved in DMSO):
  1. Weight 2.5 mg of TMRM and dissolve in 1 mL DMSO;
  2. Divide into 0.2 mL portions (use dark vials);
  3. Store at -20 °C.
Preparation of 1 mM solution - use for experiment:
  1. Dilute a 200 µL aliquot with 800 µL DMSO to reach 1 mL final volume;
  2. Divide into 0.2 mL portions (use dark vials);
  3. Store at -20 °C.


Signal and output

  1. Signal: The O2k-Fluo LED2-Module is operated through the Amperometric (Amp)-Channel of the O2k, with electric current (ampere [A]) as the primary signal.
  2. Output: The focus of the output with TMRM is on Type C: Force, mt-membrane potential.



The TMRM method can be used with the O2k-Fluo LED2-Module. For examples of the use of TMRM with mouse liver mitochondria, rat liver tissue homogenate, and permeabilized HEK 293T cells see the discussion page of this entry