TMRM: Difference between revisions
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== Application in [[HRFR]] == | == Application in [[HRFR]] == | ||
::: '''TMRM: Tetramethyrhodamine methyl ester perchlorite''' (C<sub>25</sub>H<sub>25</sub>CIN<sub>2</sub>O<sub>7</sub>); Sigma T 5428 or ThermoFischer T 668, 25 mg, store at -20ยฐC; FW = 500.93 | ::: '''TMRM: Tetramethyrhodamine methyl ester perchlorite''' (C<sub>25</sub>H<sub>25</sub>CIN<sub>2</sub>O<sub>7</sub>); ย | ||
ย | ::: Sigma T 5428 or ThermoFischer T 668, 25 mg, store at -20ยฐC; FW = 500.93 | ||
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:::'''Preparation of 5 mM stock solution (dissolved in DMSO)''': | :::'''Preparation of 5 mM stock solution (dissolved in DMSO)''': | ||
::::# Weigh 2.5 mg of TMRM | ::::# Weigh 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 ยฐC. | ::::# Store at -20 ยฐC. | ||
| Line 24: | Line 23: | ||
:::'''Preparation of 1 mM solution - use for experiment''': | :::'''Preparation of 1 mM solution - use for experiment''': | ||
::::# Dilute a 200ยตl aliquot with 800ยตl DMSO to reach 1 mL final volume | ::::# Dilute a 200ยตl aliquot with 800ยตl 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 ยฐC. | ::::# Store at -20 ยฐC. | ||
Revision as of 15:27, 30 January 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 ฮฯmt. 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 -20ยฐC; FW = 500.93
Caution: Light sensitive (store solution in a dark vial)!
- Preparation of 5 mM stock solution (dissolved in DMSO):
- Weigh 2.5 mg of TMRM and dissolve in 1 ml DMSO.
- Divide into 0.2 ml portions (use dark vials)
- Store at -20 ยฐC.
- Preparation of 5 mM stock solution (dissolved in DMSO):
- Preparation of 1 mM solution - use for experiment:
- Dilute a 200ยตl aliquot with 800ยตl DMSO to reach 1 mL final volume
- Divide into 0.2 ml portions (use dark vials)
- Store at -20 ยฐC.
- Preparation of 1 mM solution - use for experiment:
Signal and output
- 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.
- 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
