Enabling Vital Screening for Microbial Evolution Research

Microbial evolution is a crucial area of crop science research, but this aspect may not be evident. Several bacteria play a key role in defending crops from pathogens and supporting plants to absorb nutrients, like nitrogen and phosphorus, from the soil.

Such microorganisms offer a cost-effective solution for protecting and improving crops and also decrease the leaching of nutrients from fertilizers into the wider surroundings. A better understanding of microorganisms, their evolution and the traits of different types of strains represent a significant part of agricultural studies.

The VIAFLO lightweight electronic pipette and the VOYAGER automatic adjustable tip spacing pipette are ideal laboratory companions for research works where a large number of samples are required to simplify workflows and obtain reproducible results.

Dr Astrid Altamirano-Junqueira, who finished her doctoral studies on the evolution of bacterial motility in the School of Biological Studies at Reading University, explained how these advanced electronic pipettes helped in her studies.

I completed my PhD in 2019 in the United Kingdom, where I was researching microbial evolution. My focus was on microbial motility, looking at the effects of deleting the flagella master regulator gene—FleQ—in Pseudomonas fluorescens.

Dr Astrid Altamirano-Junqueira, School of Biological Studies, Reading University

During bacterial evolution, genes usually duplicate and mutate to assume new functions.

Dr Altamirano-Junqueira conducted comprehensive gene expression analysis on over 40 diverse FleQ knockout P. fluorescens strains and found a new strain that was subjected to rewiring of the nitrogen pathway, of which the flagella master regulator gene FleQ is a part in the wild type, to restore the motility and expression of flagella proteins.

Dr Altamirano-Junqueira then continued to investigate how this novel 'swimmer' strain was impacted by nitrogen starvation.

VOYAGER – Adjustable tip spacing electronic pipette

Video Credit: INTEGRA Biosciences

Enabling quick results

My first task was to grow the FleQ knockout mutants which, depending on the treatment, could take between one and 20 days to appear in the culture media.

Dr Astrid Altamirano-Junqueira, School of Biological Studies, Reading University

Since P. fluorescens is a mobile microorganism by nature, standard incubator setups developed for organisms that grow in suspension or bind to a surface did not offer a practical option; inadequate agitation of aflagellate P. fluorescens caused the bacteria to settle at the base of the well and become resource-starved.

This results in reduced growth over time, even in the absence of nitrogen starvation, possibly disrupting the outcomes. Hence, Dr Altamirano-Junqueira had to employ a Bioscreen C MBR analyzer, which allows relatively stronger plate shaking, and can make on-the-fly measurements even when the plate is not immobile and unsettled.

Enabling Vital Screening for Microbial Evolution Research

Image Credit: INTEGRA Biosciences

According to her, one major difficulty of using the Bioscreen C MBR analyzer is that it utilizes a honeycomb design microplate, which lacks the same well spacing as the regular 96-well microplates.

Initially, Dr Altamirano-Junqueira was pipetting the samples into every well separately, but it is possible to incubate and examine up to 200 specimens at a single time on this platform, rendering the procedure incredibly time-intensive.

During the first attempt, Dr Altamirano-Junqueira took nearly six hours to prepare and inoculate the plate which was quite tiring for her.

She was also concerned about the fact that even a slight lack of concentration would lead to serious errors. It proved to be equally difficult for her to keep the plate cold so that the microorganisms do not start growing before the plate was completely loaded.

Since it was a time-critical step, she actually required a more efficient solution for her pipetting tasks; hence, she began to look for alternative pipetting options and ultimately came across the multichannel electronic pipette range from INTEGRA.

As a matter of fact, the Bioscreen manual has also suggested the VOYAGER adjustable tip spacing pipette as the most effective option for use along with the instrument. The pipette makes it easy to transfer numerous specimens from tubes to plates, or even between plates with varied spacing.

With the VOYAGER adjustable tip spacing pipette, Dr Altamirano-Junqueira can now load a whole plate within 15 minutes, which according to her is quite incredible.

Enabling Vital Screening for Microbial Evolution Research

Image Credit: INTEGRA Biosciences

The perfect combination for a variety of workflows

This combination of the VOYAGER pipette and the Bioscreen C MBR analyzer has also helped Dr Altamirano-Junqueira to grow and study numerous biological replicates of multiple mutants, investigate the impacts of nitrogen starvation and also the effect of different sources of nitrogen on bacterial mobility.

She also got a single-channel VIAFLO electronic pipette for use in other facets of her workflow, and this electronic pipette combined with the VOYAGER pipette is incredibly flexible.

At present, Dr Altamirano-Junqueira is using both pipettes for an array of different jobs, such as PCR setup and gel loading. According to her, these pipettes make it relatively faster when compared to using a mechanical single-channel pipette, and since the VIAFLO is an electronic pipette, it does not put a strain on one’s thumb.

In standard manual pipettes, plenty of pressure needs to be applied to the plunger to pipette but when one is processing several specimens or conducting a process that has numerous steps, it can seriously take its toll on the individual’s hand.

The electronic pipettes decreased the chance of repetitive strain injuries, and they are also precise and fast, saving both time and money. These features had allowed Dr Altamirano-Junqueira to process more specimens in a single day.

I also love that you can program both pipettes to match your workflow; I found this very useful for my research, as I had so many strains and needed to perform multiple replicates for each one.”

Dr Astrid Altamirano-Junqueira, School of Biological Studies, Reading University

Enabling Vital Screening for Microbial Evolution Research

Image Credit: INTEGRA Biosciences

Expanding experimental capabilities

For Dr Altamirano-Junqueira, the time she saved by using the VOYAGER pipette and the VIAFLO pipettes allowed her to analyze more strains during her final year of PhD studies, and also helped her to study more culture conditions. This allowed her to gain maximum information about the strains, which was crucial for multivariate analysis.

Dr Altamirano-Junqueira was able to actually study statistics and gain a better understanding of the various mutants. She was also able to detect a new ammonia transporter that, post-genome sequencing, resulted in the discovery of a novel strain, called FleQS4.

At present, this is in the process of publication, and because of INTEGRA’s electronic pipettes, she was able to carry out all the PCR experiments as well as screening for this specific gene, for all the strains, in just a matter of two weeks. Dr Altamirano-Junqueira expressed her thanks for the VOYAGER pipettes.

Enabling Vital Screening for Microbial Evolution Research

Image Credit: INTEGRA Biosciences

About INTEGRA Biosciences

INTEGRA provides innovative solutions for Liquid Handling and Media Preparation applications which serve the needs of their customers in research, diagnostics and quality control laboratories.

Their instruments and plastic consumables are developed and manufactured in Zizers, Switzerland and Hudson, NH USA. In order to remain close to their customers, they maintain a direct sales and support organization in North America, the UK, France and Germany, as well as a network of over 100 highly trained distribution partners worldwide.

In recent years they have focused on developing a new and technologically advanced range of handheld electronic pipettes which are simple to use and meet the ergonomic needs of their customers. Today they are proud to offer the widest range of electronic pipettes in the market spanning a range from single channel pipettes up to 384 channel bench-top instruments.

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Last updated: Mar 3, 2021 at 3:46 AM


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