Bioluminescence Measurements

Fluorescence and bioluminescence measurement of cytoplasmic free calcium 16... 

Ito, K. Nakagawa, K. Murakami, S. Arakawa, H. Maeda, M. Highly sensitive simultaneous bioluminescent measurement of acetate kinase and pyruvate phosphate dikinase activities using a firefly luciferase-luciferin reaction and its application to a tandem bioluminescent enzyme immunoassay. Anal. Sci., 19, 105-109 (2003)... 

Other Parameters Measured Simultaneously with Bioluminescence. Measurements of several other parameters were obtained from the seawater after it had traversed the bioluminescence detector. When working on station with the bathy-photometer, which was equipped with a pressure transducer, temperature and beam transmittance were measured at depth while seawater was pumped by the submersible pump at depth to shipboard with 110 m of 2.54-cm ID hose. Sea surface temperature was obtained continuously from a probe at the intake near the sea chest. The seawater, obtained from either the sea chest or the bathyphotometer, was pumped through a Turner Designs fluorometer to measure chlorophyll fluorescence, and past a pH probe (31) and a conductivity cell when available. Samples of seawater were frozen for subsequent nutrient analysis (NO , NH4OH, P04 , and NO2). Plankton filtrates from 20 to 100 L (depending on plankton abundance) of seawater were collected from a 100-L effluent tank fitted with plankton net collection cups of 20-fxm mesh porosity. The filtrate was split, filtered onto Whatman GF/C 4.25-cm filter discs, and frozen for subsequent carbon and nitrogen determinations. The other half of the sample was preserved in 5% buffered formaldehyde solution for taxonomic analysis. 

Data obtained from the pumped (closed) deep bathyphotometer and the open bathyphotometer mounted on the Alvin submersible show that skylight penetrates to 200 m on a moonless night and to 500-700 m in daylight conditions. Therefore, bioluminescent measurements at depths to about 200 m at night and 500 m in the daylight can reasonably be done only with a closed system. (These depths can vary considerably with the turbidity of the seawater.)... 

R6. Roda, A., Kricka, L. J., DeLuca, M., and Hofmann, A. F., Bioluminescence measurement of primary bile adds using immobilised 7a-bydroxy-steroid debydrogenase Application to serum bile adds. /. lipid Res. 23, 1354—1361 (1982). 

Bartholomeusz DA, Andrade JD. Photodetector calibration method for reporting bioluminescence measurements in standardized units. Luminescence 2002 17 77-115. 

Instrumentation Luminometer 3550i and Filtravette (New Horizons Diagnostics Corp., USA) were used for bioluminescent measurement. 

After pretreatment proposed it was possible to filter up to 1 mL of force-neat suspension through Filtravette , but the optimal volume for bioluminescence measurement was 0.1 mL. In that case the residual non-bacterial ATP concentration... 

Luminometer 3550i and Filtravette (New Horizons Diagnostics Corp., USA) were used for bioluminescent measurement. ATP-reagent (based on soluble Luciola mingrelica firefly luciferase) was developed in our laboratory. Dimethyl sulfoxide (DMSO) was from Reakhim (Russia). Nutrition broth (NB) from ICN, Tryptic Soy Broth (TSB) from Difco and Thioglycolate broth (TB) from Merck were used. Ultrapure deionized water was obtained on Milli-Q (Millipore). Broth culture of Escherichia coli LE392 was used in model experiments. 

Portable PMT-based luminometer LUM-1 used for bioluminescence measurement was from Lumtek LLC (Russia). The 48- and 96-well multidishes were from Nuclon . The 96-well strip plates were from Nunc. 

Sensitive methods for SBA based on bioluminescence measurement have recently been deve p using hydroxysteroid dehydrogenase (3o( or 12d or 7d -HSD). The HSD catalyzes the conversion of the bile acid hydroxy g up (3o( or 7d or 12 c. ) to a keto-group in the presence of NAD(P). The resulting NAD(P)H, in presence of NAD(P)H FMN oxydoreductase (OXRED), converts FMN to its reduced form (FMNH ). This in turn reacts with decanal and oxygen in the presence dr bacterial luciferase to produce light. The intensity of the light emitted is proportional to the amount of bile acids in the initial raction. The overall scheme,for 7 -HSD for instance, is this ... 

Cobbold PH and Rink TJ (1987) Fluorescence and bioluminescence measurement of cytoplasmic free calcium. Biochemical Journal 248 313-328. 

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