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Working in BSL-3/4 laboratories

How can remote monitoring of cell cultures improve safety and efficiency?

As the World undertakes the battle against the Covid-19 pandemic, most of the public probably think working with dangerous viruses could be very exciting, that the daily life of scientists is full of adrenaline like in the movies.

However, we scientists know that there is another side to the story: an arduous and potentially dangerous side.

Why? Put simply, it’s an issue with getting dressed.

Handling pathogenic microorganisms like SARS-CoV-2, Ebola, or Mycobacterium tuberculosis has to be done in high containment laboratories. Working in BSL-3 or BSL-4 (biosafety level 3 or 4) laboratories, procedures which are usually simple in your average lab become more time consuming and demanding, because researchers have to dress up (or overdress) in a specific way, follow a long list of rules.

The work carried out in these labs is vital for the understanding of highly pathogenic viruses or bacteria, so making the process more efficient and user friendly should be one of a lab manager or PI’s main priorities.


Attention to researchers working on COVID-19:

We are currently donating remote monitoring devices for BSL-3/4 laboratories working to combat COVID-19.

Learn more : CytoSMART Lux2 for supporting the fight against covid-19



The challenges of working in a BSL-3/4 laboratory

The work that takes place in BSL-3 and BSL-4 labs is vital, but potentially dangerous, hence the high containment environment. The rationale behind this approach is that increased containment protects the user and the community from possible exposure to the pathogen. In short, protecting the team involved and the wider world.

This high level protection is only achieved when direct contact with the microorganism is minimized. Previously the most effective ways to achieve this has been by using special personal protective equipment, filtering the air circulating the lab, having a negative pressure to draw the airflow into the laboratory, decontaminating all objects that exit, among others.

These protective conditions have been absolutely necessary to make the work in BSL-3 and BSL-4 laboratories safe, but in turn this increases workloads and makes processes more demanding.

One example which illustrates how onerous the safety processes in these types of laboratories can be, is the entering procedure of a BSL-3:

  1. Enter the dressing room
  2. Put the first set of protective gloves on
  3. Add disposable shoe covers
  4. Put on either the first lab coat (or overall)
  5. Move into the clean area
  6. Add a second lab coat
  7. Apply a hair cover
  8. Put some PPE goggles on
  9. Pop your protective face mask into place
  10. Add a second pair of gloves
  11. Finally, enter the lab and start monitoring the cells.

If everything goes well, this 11 (yes 11!) stage procedure can take up to 12 minutes. If you’re entering a BSL-4 then double that time as users of these labs are required to completely change their clothing before entering and to shower upon exiting.

Sounds like fun? Honestly, it really can be. Working in these laboratories gives you a completely unique opportunity to handle the active and infectious form of your pathogen of interest, and of course this must be done in a safe, controlled manner. But in addition to it being fascinating, working in these conditions can be:

  • laborious
  • time consuming
  • expensive
  • damaging to the environment (a lot of disposable material is required).

Because of this, users of BSL-3 and BSL-4 laboratories must plan their experiments in such a way that only the most necessary steps are carried out in the lab, usually limiting the number of times the personnel enters and the time spent inside.

But there is another way…

Remote monitoring of cell cultures is a viable, effective option

A creative alternative to reduce the time spent in a BSL-3 or BSL-4 laboratory is the introduction of remote monitoring of cell cultures in incubators. Remote monitoring simply means assessing the cells and monitoring them through a live online environment, with no need to remove them from the incubator or enter the lab.

To monitor cells in this way, a small microscopy platform is set up inside the CO2-incubator, allowing cells contained in a dish or flask to be monitored in real time over extended periods of time. CytoSMART, for example, has developed bright-field microscopy systems that operate from right inside standard incubators. These microscopes offer the possibility of performing time-lapse videos of 2D cell cultures grown in standard dishes.

For standard cell culture monitoring we recommend the CytoSMART Lux2

For more advanced solutions the CytoSMART Omni can be considered. This microscope can be used to monitor culture vessels that are used by production facilities to obtain high yields. The example below explains how the CytoSMART Omni is used to image the complete area of a HYPERFlask to assess confluency.


Sounds great, what are some practical applications in BSL-3 and BSL-4 labs?

How can remote monitoring of cell cultures be used in high containment facilities? is a question we’re oftened asked and we have many examples. For instance, a typical experiment done in a BSL-3 or BSL-4 lab could be to characterize the infection of host cells by the pathogen in question. Upon infection, a permissive cell changes its morphology (detaches, forms syncytium, shrinks), behavior (stops dividing, decreases motility) and, sometimes even dies.

While the viruses or intracellular bacteria cannot be easily seen by the naked eye, the effect that the infection has on the cell as a whole can be assessed in terms of cytopathic effects. With a remote monitoring system, a kinetic evaluation in which the cytopathic effect is followed and recorded over time could be performed without having to enter the laboratory at every given time point.

Even more impressive, a remote monitoring system can be used to study the effect of antiviral or antibacterial drugs on infection in these cell cultures. By monitoring cell proliferation, motility or morphology over time, the following questions can be answered:

  • Can compound A prevent cell death induced by the pathogen?
  • Can compound B make the growth rate of infected cells comparable to the one of not-infected ones?
  • Is there a cytotoxic effect of compound C?

To conclude…

Working in a high containment laboratory like a BSL-3 or BSL-4 may not be quite as exciting as it seems on your favourite TV drama. As a user, you’re likely to be spending a good portion of you time getting ready to work in the lab and considering all the necessary measures required to keep you safe.

A practical, effective alternative such as remoting monitoring, allows vital work to continue whilst saving time and maintanining the most basic principle of high containment laboratories: avoiding contact.



Author:

Vanesa Ayala-Nunez, PhD

Vanesa is a Research Scientist with more than ten years of experience in the field of infectious diseases. Working at the interface of Virology, Immunology, and Cell Biology, she has dealt with pathogens like HIV, Zika, dengue and Chikungunya. She is currently based in the sunny corner of Germany.



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