Overview

The initial training in the laboratory area plays a vital role in fostering a harmonious working environment and ensuring the effective integration of the techniques which are applied in our experiments. Acquiring knowledge of the rules and safety protocols not only helped to prevent accidents but also safeguard the integrity of the results we generate. It is crucial, for instance, to be aware of the designated work areas for handling specific substances or enzymes, as well as to properly dispose of materials to avoid any potential contamination within the workspace. Furthermore, the laboratories that support our research endeavor are equipped with state-of-the-art facilities and resources tailored to meet our specific needs. This comprehensive setup significantly contributed to the smooth and successful execution of our experiments.

Lab safety

Safety Features of Lab

  1. Chemical fume hood A fume hood is a type of local ventilation device that is designed to limit exposure to hazardous or toxic fumes, vapors or dusts. The chemical fume hood played an indispensable role in our experimental setup. Its importance stemmed from the need to ensure the safety and environmental responsibility of our procedures. Specifically, the utilization of ozone drying for cleaning the gold biosensors, known to produce inhalation hazards and atmospheric emissions, necessitated the controlled containment afforded by the fume hood. Additionally, throughout the preparation of magnetic beads and the mixing and dissolution of FeCl2 and FeCl3 in deionized water (dH2O), the fume hood served as a vital measure and this step was imperative, given that the reactions entailed the generation of gaseous byproducts that poses inhalation risks and mandated rigorous containment.
  2. Fire extinguisher
  3. Waste Disposal “Waste is a fact of Lab life, but luckily there are tons of super knowledgeable health and safety officers to tell us what to do with all of it!” Useful video link: How To: Lab Waste
  4. First Aid Kit

Safety training

There were two laboratories where the experiments took place. One laboratory is housed in the Department of Biology of the University of Crete, while the Dr. Fillipidi’s Lab, which functioned as a secondary laboratory, is housed in the Institute of Molecular Biology and Biotechnology (IMBB- FoRTH). In order to be able to work in these areas, our supervisor Dr. Filippidi informed us of the safety rules that are followed without exception by everyone, as well as what we can do in the event of an emergency. At Prof. Gizeli’s lab, our instructor, Dr. Chronaki showed us where the waste disposal is, how and when we have to use the chemical fume hood and what to do also, in a case of emergency.

Cell-free system or Cell-based system

A cell-free system is an experimental setup that does not involve intact cells. Instead, it uses cell extracts or purified cellular components to study specific biological processes or reactions in a controlled environment outside of a living cell. In contrast, a cell-based system involves using whole living cells as the basis for experiments or research. These cells can be of various types, including bacterial, yeast, plant, or animal cells, depending on the specific research goals. Both approaches have their unique advantages and applications in scientific research and biotechnology. However, appreciating the positive characteristics of the cell-free system, we aimed to use one with the aim of pioneering and ensuring that issues of ethical conduct and laboratory safety are respected.

Βriefly some features of the cell free system:

  1. Cell-free systems are more accessible than working with living cells. They do not require extensive facilities or specialized equipment for cell culture, making them an attractive option for many laboratories.
  2. Each researcher has the possibility to adjust the pH, the temperature or even the solvent to use regardless of the needs of cells, but serving the optimal conditions for the detection goal.
  3. Cell-free systems can operate in the presence of toxins that would inhibit or kill live cells. This means that key sensing and metabolic components, such as transcription factors and enzymes, can be produced in higher concentrations than in living cells, leading to improved sensitivity and efficiency.
  4. Cell-free systems enable rapid experimentation and screening of reactions or compounds.

Working with pesticides

Deltamethrin (delta)

Deltamethrin is an insecticide belonging to the pyrethroid family. Pyrethroids are the man-made versions of pyrethrins, natural insecticides from chrysanthemum flowers. Deltamethrin serves as an insecticide and veterinary drug and is extensively used to control pests worldwide. With the increasing use of deltamethrin, its toxic effects on animals and humans have attracted great attention. Unfortunately, as its usage increased, so has the number of poisoning cases associated with deltamethrin. Fortunately, the mortality in deltamethrin poisoning cases is low.

How might I be exposed to deltamethrin?

You can be exposed to deltamethrin(delta) if you touch, eat, or breathe it in. As an example, it could be breathed in if a fine mist or dust containing deltamethrin gets in the air you breathe. Exposure to deltamethrin can be limited by reading and following label directions. Whenever we used delta dust we wore a mask and gloves while working carefully on a bench so it wouldn't spread. Finally, we used quite small concentrations throughout the experimental design.

Working with aptamers

Aptamers are single-stranded DNA or RNA molecules which, due to their unique three-dimensional properties, can bind their target with high affinity. In our project what is used is one of the most common aptasensor formats, the duplexed aptamer (DA). DAs are aptasensors that contain two nucleic acid elements coupled via Watson–Crick base pairing. One is an aptamer sequence, which serves as a ligand-specific receptor, and the other is an aptamer-complementary element (ACE), such as a short DNA oligonucleotide, which is designed to hybridize to the aptamer. The ACE competes with ligand binding, such that DAs generate a signal upon ligand-dependent ACE-aptamer dehybridization. Some features of aptamers that led to their selection as our key molecules are described below:

  1. Compared to antibodies, the aptamer is more thermal stable, easier to be modified, conjugated, and amplified.
  2. They can be used in cell-free systems, avoiding culturing or even modifying cell lines to convert them as biosensors.
  3. Compared to other molecule-based assays or diagnostic technology, aptasensors are simple, rapid, less expensive, and can be used in real time.
  4. 4. The fact that they can detect such small amounts of delta leads to a reduced concentration of it during the experimental process.

Human practices safety- Ethics

When we refer to safety we may immediately think of a laboratory, but the term does not extend only there. A very important condition is the safety of human rights. At every activity involving children or adults, we had permission to take photos, record views and share information. Especially regarding minors, we asked for permission from the parents attending the venue. Beyond the protection of intellectual rights and more focused on the part of ethics and how it affects or influenced by science, an open discussion was held on the topic 'Is it ethically justifiable to harness and, potentially, induce some degree of environmental degradation in the pursuit of bolstering productivity and enhancing the quality of human life?' This thought-provoking dialogue featured a panel comprising four esteemed guest speakers, alongside a diverse assembly of participants. The discussion was thoughtfully structured, featuring a deliberate division into thematic segments to facilitate a comprehensive exploration of this multifaceted issue.