Mitochondrial Metabolism

One of the drawbacks of optical spectroscopy, affecting absorbance in particular, is the effect of light scattering on the measurement. A highly scattering sample will increase the apparent absorbance in the vast majority of spectrophotometers on the market. The instrument cannot differentiate between scattered and absorbed light. In fact, this artifact is commonly used to roughly quantitate bacterial cell cultures by measuring the apparent absorbance at 600 nm.

Turbid Samples

In the 1980’s, this problem was partially addressed with the introduction of the Aminco DW2 and later the DW2000. Olis still supports these amazing optical benches. However, Olis has now topped this design with the CLARiTY line of instruments, which have the ability to record true absorbance spectra in the presence of a highly scattering medium. The Olis CLARiTY line of instruments are ideally suited for monitoring absorbance changes in mitochondrial suspensions, whole cells, or other highly scattering environments.

The CLARiTY line includes the RSM 1000, DM 245, and the DB 620 optical benches. The RSM 1000 configuration allows for the collection of rapid scans and is usually configured to measure up to 100 scans per second. The DM 245 and DB 620 are configured for single wavelength measurements over time or for conventional absorbance scans. The CLARiTY chamber includes magnetic stirring and temperature control from 25 to 80 C˚. Common accessories include the TLC 50 (for RSM 1000), StepDisks (for RSM 1000), Twin Peltier (for DW2/2000), and fluorescence module (for DW2/2000).

Client Publications
CL Quinlan, RLS Goncalves, M Hey-Mogensen, N Yadava, VI Bunik, MD Brand. (2014). The 2-Oxoacid Dehydrogenase Complexes in Mitochondria Can Produce Superoxide/Hydrogen Peroxide at Much Higher Rates than Complex I.
      J. Biological Chemistry DOI: 10.1074/jbc.M113.545301 jbc.M113.545301
Achieved using the OLIS DW 2

IV Perevoshchikova, CL Quinlan, AL Orr, AA Gerencser, MD Brand. (2013). Sites of superoxide and hydrogen peroxide production during fatty acid oxidation in rat skeletal muscle mitochondria.
      Free Radical Biology and Medicine 61C, 298-309
Achieved using the OLIS DW 2

S Smith, A Witkowski, A Moghul, Y Yoshinaga, M Nefedov, P de Jong, D Feng, L Fong, Y Tu, Y Hu, SG Young, T Pham, C Cheung, SM Katzman, MD Brand, CL Quinlan, M Fens, F Kuypers, S Misquitta, SM Griffey, S Tran, A Gharib, J Knudsen, HK Hannibal-Bach, G Wang, S Larkin, J Thweat, S Pasta. (2012). Compromised Mitochondrial Fatty Acid Synthesis in Transgenic Mice Results in Defective Protein Lipoylation and Energy Disequilibrium.
      PLoS ONE 7(10): e47196
Achieved using the OLIS DW 2

M Gutierrez-Aguilar and S Uribe-Carvajal. (2015). The mitochondrial unselective channel in?Saccharomyces cerevisiae.
      Mitochondion DOI: 10.1016/j.mito.2015.04.002
Achieved using the OLIS SLM 8000

IG Shabalina, M Vrbacky, A Pecinova, AV Kalinovich, Z Drahota, J Houstek, T Mracek, B Cannon, J Nedergaard. (2014). ROS production in brown adipose tissue mitochondria: The question of UCP1-dependence.
      Biochimica Biophysica Acta - Bioenergetics DOI: 10.1016/j.bbabio.2014.04.005
Achieved using the OLIS DW 2

RLS Goncalves, CL Quinlan, IV Perevoshchikova, M Hey-Mogensen, MD Brand. (2014). Sites of Superoxide and Hydrogen Peroxide Production by Muscle Mitochondria Assessed ex Vivo under Conditions Mimicking Rest and Exercise.
      J. Biological Chemistry 290, 209-227
Achieved using the OLIS DW 2000

IG Shabalina, AV Kalinovich, B Cannon, J Nedergaard. (2015). Metabolically inert perfluorinated fatty acids directly activate uncoupling protein 1 in brown-fat mitochondria.
      Archives of Toxicology DOI:?10.1007/s00204-015-1535-4
Achieved using the OLIS DW 2