Oxygraph chamber

Manufactured by Hansatech

Sourced in United Kingdom

The Oxygraph chamber is a laboratory equipment used for the measurement of oxygen consumption or production in biological samples. It provides a sealed environment to monitor changes in oxygen levels over time.

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Lab products found in correlation

Enliten atp assay system, by Promega (1 mentions) Clark type electrode, by Hansatech (1 mentions) Bca protein reagent, by Thermo Fisher Scientific (1 mentions)

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Oxygraph

3 protocols using oxygraph chamber

Photosynthesis and Respiration in U. compressa

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Photosynthesis was quantified cultivating 25 mg of fresh tissue (FT) of U. compressa in 2 mL of seawater in an oxygraph chamber (Hansatech, Norfolk, UK). O₂ production was detected for 10 min, under a light intensity of 425 μmoles m^− 2 s^− 1 and respiration was determined cultivating 25 mg of the alga (FT) in 2 mL of seawater in the oxygraph chamber, and O₂ consumption was detected for 10 min in darkness.

Laporte D., Rodríguez F., González A., Zúñiga A., Castro-Nallar E., Sáez C.A, & Moenne A. (2020). Copper-induced concomitant increases in photosynthesis, respiration, and C, N and S assimilation revealed by transcriptomic analyses in Ulva compressa (Chlorophyta). BMC Plant Biology, 20, 25.

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Oxygen Consumption and ATP Analysis

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A6MT cells were grown for 24 h in gal-DMEM or in gal-DMEM supplemented with 6 mM DMK. Cells were collected by trypsinization, pelleted by centrifugation, and resuspended in the same culture medium. Cell suspension was transferred in an Oxygraph chamber equipped with Clark-type electrode (Hansatech) where oxygen consumption was measured at 37°C as described (D’Aurelio et al., 2001 (link)). Alternatively, oxygen consumption was measured in cybrids with various proportion of COXI mutation, as described above, in DMEM without glucose containing 1 mM pyruvate and 4 mM glutamine. Alternatively, oxygen consumption was measured in A6MT and WT digitonin-permeabilized cells using 6 mM pyruvate or 6 mM αKG as substrates, 200 nmoles ADP (0.7 mM) for rapid state 3 respiration, and 1 mM KCN as terminal inhibitor, as described (Hofhaus et al., 1996 (link)).
ATP content was measured using an Enliten ATP assay system (Promega), according to the manufacturer’s instructions.
The activity of complex IV was measured in isolated tissue mitochondria as described previously (D’Aurelio et al., 2010 (link)).

Chen Q., Kirk K., Shurubor Y.I., Zhao D., Arreguin A.J., Shahi I., Valsecchi F., Primiano G., Calder E.L., Carelli V., Denton T.T., Beal F.M., Gross S.S., Manfredi G, & D’Aurelio M. (2018). Rewiring of glutamine metabolism is a bioenergetic adaptation of human cells with mitochondrial DNA mutations. Cell metabolism, 27(5), 1007-1025.e5.

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Quantification of Cyanobacterial Biomass

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Cell dry weight concentrations were measured directly as ash- free dry weight (AFDW) (Pinchuk et al., 2010 (link)) and compared in a standard curve to the indirect OD₇₃₀ measurements obtained using a Genesys 20 visible spectrophotometer (Thermo Scientific, Rockford, IL). Protein was quantitated using BCA protein reagent (Thermo Scientific). Chl a and phycocyanin concentrations were estimated using a previously described method that corrects the effect of scatter on absorbance from whole-cell suspensions (see Supplemental Methodologies) (Myers, 1980 ; Burns et al., 2006 (link)). Dissolved O₂ concentration in the reactor was measured with a Clark O₂ electrode (InPro^® 6800Series, Mettler Toledo International Inc., Columbus, OH). O₂ production rates as a function of “white light” irradiance (tungsten incandescent) were measured inside an oxygraph chamber (Hansatech, Norfolk, UK).

Bernstein H.C., Konopka A., Melnicki M.R., Hill E.A., Kucek L.A., Zhang S., Shen G., Bryant D.A, & Beliaev A.S. (2014). Effect of mono- and dichromatic light quality on growth rates and photosynthetic performance of Synechococcus sp. PCC 7002. Frontiers in Microbiology, 5, 488.

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