Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

Abstract

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

Live-Cell Systems in Real-Time Biomonitoring of Water Pollution: Practical Considerations and Future Perspectives

“…This is, in turn, reflected by the detectable changes in electrical impedance. The ECIS technology allows a label-free, non-destructive and real-time analysis of cell viability and proliferation [141][142][143]. It provides several analytical advantages such as (i) a lack of any fluorescent labels and imaging, (ii) the ability to perform real-time kinetic monitoring of cells, (ii) ease of operation and (iv) straightforward automation [140,141].…”

“…The utilisation of vertebrate cell biosensors including ECIS technology for the monitoring of water quality has, thus far, been a niche application with only a very small number of prototypes presented as compared to bacterial and algal sensors [139,140,143,144]. The theoretical advantages of mammalian cell biosensors include a real-time response to a wide variety of toxicants in real-time.…”

“…This is advantageous when compared to cell-free biosensing methods such as immunosensors, enzymatic and DNA biosensors. When utilising human cell lines, one can also postulate that their responses might also be better applicable to modelling the potential impact of toxicants on human health [143].…”

Altered function of the left ventricle and clinical significance of heart‑type fatty acid‑binding protein in cardiac dysfunction among patients with sepsis

“…Of note, cardiac dysfunction usually occurs in sepsis patients, which gives rise to reduced systemic perfusion of the organ and in turn enhances disease progression ( 5 , 6 ). For patients who die due to septic shock and severe sepsis, most succumb due to damage to the cardiovascular system, and ~40% of patients who survive sepsis develop myocardial damage ( 7 ). Furthermore, these conditions worsen when cardiac dysfunction occurs earlier, along with a higher death rate ( 8 ).…”

A portable chemical toxicity biochip based on electronic enzymatic monitoring of cytotoxic effects on fish cells

“…However, one of the major challenges in the development of portable mammalian cell biosensors for on-site analysis is the requirement for growth at 37 • C in a 5% CO 2 atmosphere and the need for frequent medium changes and subculture [20]. As an alternative, a number of groups have employed fish cells as a biological indicator [21][22][23][24] with electric cell substrate impedance sensing (ECIS) or amperometric measurement of redox mediators used to assess changes in cell viability upon toxicant exposure. Brennan et al (2012) developed an ECIS biochip that employed rainbow trout gill cells (RTgill-W1) as the biological recognition element [25].…”