Bioluminescence alternatives

Gandelman OA, Church VL, Moore CA, Came C, Jalal H, Murray JAH, Tisi LC. BART - bioluminescent alternative to real-time PCR. In Szalay AA, Hill PJ, Kricka LJ, Stanley PE. eds. Bioluminescence and Chemiluminescence. Chemistry, Biology and Applications. Singapore World Scientific, 2007 95-8. 

Fig. 8.9 Possible mechanisms of the bioluminescence reaction of dinoflagellate luciferin, based on the results of the model study (Stojanovic and Kishi, 1994b Stojanovic, 1995). The luciferin might react with molecular oxygen to form the luciferin radical cation and superoxide radical anion (A), and the latter deproto-nates the radical cation at C.132 to form (B). The collapse of the radical pair might yield the excited state of the peroxide (C). Alternatively, luciferin might be directly oxygenated to give C, and C rearranges to give the excited state of the hydrate (D) by the CIEEL mechanism. Both C and D can be the light emitter.

Branchini, B. R., et al. (2004). An alternative mechanism of bioluminescence color determination in firefly luciferase. Biochemistry 43 7255-7262. 

Gautier S., Blum L.J., Coulet P.R., Alternate determination of ATP and NADH with a single bioluminescence-based fiber-optic sensor, Sensor Actuat B-Chem 1990 1 580. 

Since the reaction efficiency in micro channels depends on the achieved mixing performance, specific structures or surface modifications are widely used to enhance mixing. Because the developed micro pumps can be controlled very easily, dynamic or alternated pumping of the fluids is possible. To monitor the capability of the system in terms of mixing speed and quality, a bioluminescence reaction is used ... 

Figure 4.19 (A) Time courses of the bioluminescence at three measurement points (2, 10, 46 mm) downstream of the Y-shaped junction. The measurement was started immediately after flow stopping. (B) Typical pattern of the driving signal for alternate pulse flow. (C) Photograph of the alternate pulse flow generated by switching at 3 Hz. This frequency is selected for the visualization of thin skins, which enhances the diffusion-based mixing along the flow axis. (D) Mixing performance of the alternate pulsed flow at a frequency ranging from 1 Hz to 1 kHz. The intensity of bioluminescence is measured at points A ( ) and B ( ) [72] (by courtesy of RSC).

Several years ago it was noted that the behavior of dioxetanes substituted with large resonance groups (for example the dioxetanes postulated as intermediates in many bioluminescent reactions) is quite distinct from that of alkyl-, alkoxy-, or simple aryl-substituted dioxetanes (T. Wilson, 1976). The decomposition products of these dioxetanes are often highly fluorescent, possessing lowest excited ttti states, the yields of excited singlet states are often remarkably high, and the dioxetane stability is apparently low. It was suggested at that time that an alternative mechanism, perhaps a truly concerted mechanism, was operative. 

Transgenic bacterial biosensors. Systems such as the Microtox assay detailed earlier use the marine species Vibrio fischeri as the sensor. Because it uses a marine bacterium, Microtox must be conducted in saline solution, which is ecologically irrelevant for most soils. Because no naturally luminescent soil bacteria are known that could be used as an alternative, one solution is to fuse the genes responsible for bioluminescence into soil-dwelling strains using recombinant technology (Paton et al., 1997). Two approaches can be used ... 

F Kohen, M Pazzagli, M Serio, J deBoever, D Vanderkerckhove. Chemiluminescence and bioluminescence immunoassay. In WP Collins, ed. Alternative Immunoassays. New York Wiley, 1985, p 103. 

A promising flash method was proposed by Lappalainen etal. (1999), applying a new measurement technique. Bioluminescence is determined immediately after Vibrio fischeri is exposed to the soil slurry and again after an incubation period of less than one minute. The first value serves as reference, assuming that the bacteria are not yet affected by the toxicants in the sample. The second value, expressed relative to the first value, indicates the effect of the toxicant. This procedure provides an alternative for coloured and turbid samples. 

This ring system has also been called l,3a,6-triazaindene and numbered as in formula 1. A third alternative numbering system (2) has also been used on one occasion. Partly reduced derivatives of imidazo[l,2-a]pyrazine have been referred to as imidazo[l,2-a]piperazines and others, as imidazolino[l,2-a]pyrazines. The majority of papers on this heterocycle are of recent origin and concern the structure and oxidation products of Cypridina luciferin, a bioluminescent substance isolated from Cypridina hilgendorfii, a small crustacean. 

Today s systems are in most cases based on the bioluminescence with ATP and luciferase from the firefly. As an alternative system it is also possible to use a colour test nicotinamide adenine dinucleotides (NAD/NADH) and nicotinamide adenine dinucleotide phosphates (NADP/NADPH), which are also compounds used for the energy transfer in the metabolism in living cells or compounds found in food debris. 

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