A microfluidic in situ analyzer for ATP quantification in ocean environments

Abstract

We have developed and tested a functionally integrated in situ analyzer, the IISA-ATP system, for microbial activity assays based on a quantitative determination of the total (particulate and dissolved) ATP in ocean environments. The IISA-ATP utilizes a PDMS-glass hybrid microfluidic device as its core functional element, which can perform cell lysis and total ATP quantification by a luciferin-luciferase bioluminescence assay in situ. Transparent heaters and a temperature sensor fabricated on a glass substrate provide temperature control. As a result of the evaluation using the microfluidic device with ATP standard solutions, the bioluminescence intensity was linearly correlated with 2 × 10(-12) to 2 × 10(-8) M of ATP. A detection limit of 1.1 × 10(-11) M was determined using the completed IISA-ATP system, which includes a miniature pumping module and a control module. As a result of the evaluation using the environmental seawater sample collected from Tokyo Bay, Japan, 2.7 × 10(-10) M of total ATP was successfully determined in the laboratory by the IISA-ATP. The system was operated at a shallow submarine hot spring area in Okinawa, Japan for an in situ trial. The result shows the system was successfully operated in situ and the total ATP was determined to be 3.4 × 10(-10) M.

Advances in Microfluidic Materials, Functions, Integration, and Applications

“…Detection was based on total microbial ATP concentration probed by luciferin–luciferase reaction using serpentine microchannels to facilitate mixing. 362 …”

Evaluation of ATP measurements to detect microbial ingress by wastewater and surface water in drinking water

“…A fully automated in situ ATP analyser has been developed for measuring ATP in sea water (Fukuba et al, 2011). Further, a fully automated sensor prototype based on the ATP assay has been developed and investigated for monitoring ATP in drinking water (Vang, 2013).…”

Micro Total Analysis Systems: Fundamental Advances and Applications in the Laboratory, Clinic, and Field

“…261 Another device for an underwater bioluminescence assay of microbial activity was successfully operated at a shallow hot spring in Okinawa (Figure 10b). 262 The microfluidic device, electronics and photomultiplier tube were contained in a pressurized vessel and waste was collected rather than dispensed into the environment. In another system, acoustic radiation was used to collect and trap microorganisms in a microfluidic device at a depth of 1200 meters.…”

“…(b) A pressurized vessel containing a photomultiplier tube, electronics, and a microfluidic device for measuring microbial activity in a shallow hot spring. Reproduced from ref 262 with permission of The Royal Society of Chemistry. (c) A microfluidic device for voltage clamp studies of oocytes during parabolic flight.…”