Proteomics Lab Uses Nitrogen Purge System to Purge O2 from Solution

 
The Trinkle Lab, founded by Dr. Laura Trinkle-Mulcahy, is a proteomics lab located in Ottawa, Canada that is focused on harnessing therapeutic strategies of targeted phosphatase activity. Protein phosphatase 1 (PP1) is an active enzyme that plays an important role in glycogen metabolism, cell progression & division, muscle contraction, and neuronal activities. Dysfunctional PP1 is associated with many diseases such as Alzheimer's, Herpes, and HIV-1 transcription.

As an accomplished cell biologist, Dr. Trinkle-Mulcahy's end goal is to target protein phosphatase 1 (PP1) with a high enough specificity to develop therapeutic strategies that can minimize these off-target effects. The lab utilizes an expansion microscopy protocol, which Dr. Trinkle-Mulcahy explains below.

"Expansion microscopy (ExM) is a sample preparation technique for super-resolution imaging in which fluorescently-labeled cells or tissues are embedded in a cross-linked network of swellable polyelectrolyte hydrogel. Addition of water isotropically expands the sample up to 10-fold, increasing the physical distance between biomolecules while preserving their spatial organization relative to each other. Our lab uses the X10 protocol to expand cultured human cells for nanoscale imaging of both nuclear and cytoplasmic structures at which the protein phosphatase PP1 plays key roles.

Molecular oxygen can inhibit polymerization of ExM gels, so a key step in the protocol is to purge the O₂ in the gelatin solution by bubbling with N2 gas. This is most easily done using a nitrogen evaporator, as it offers a controlled bubbling rate. Our lab uses an N-EVAP Nitrogen Evaporator (Organomation) for this step."

The lab originally purchased Organomation's parallel evaporator for its typical use of concentrating liquid solvents ahead of processes like chromatography or mass-spectrometry. However, they have found great success in using the unit to purge oxygen from their gelatin solutions. Dr. Laura Trinkle-Mulcahy stated, "We were very happy that it [N-EVAP nitrogen evaporator] was available to us when we needed it for this more unique application."

"We were very happy that it [N-EVAP nitrogen evaporator] was available to us when we needed it for this more unique application."

The gas flow can be adjusted at each individual position, allowing the user to have precise control over the amount of gas being bubbled into each gel sample, protecting their integrity. The height of the gas distribution needles is also able to be adjusted at each sample position. This is especially helpful for the Trinkle Lab when they move onto the second round of purging which requires the gelatin samples to be actively chilled to 4 °C. 

The evaporator's design allows them to raise the needle assembly high enough to fit a beaker filled with ice water under it. The sample is placed inside the beaker to cool, at which point the needle assembly can be lowered into the gelatin to finish purging. The lab has truly been able to utilize the flexibility and versatility that the N-EVAP nitrogen evaporator offers.

For more information on the Trinkle Lab and their work, visit www.trinklelab.com. 

Do you have your own unique application that involves the use of an Organomation evaporator or extractor? Reach out to sales@organomation.com for your chance to be a featured laboratory on our blog!

References

1. Truckenbrodt et al Nat Protoc 14, 832-863 (2019)
2. Truckenbrodt et al EMBO Rep 19, e45836 (2018)
3. Gaudreau-Lapierre et al STAR Protoc 2, 100630 (2021)
4. Do et al iScience 23, 101664 (2020)
5. Mehta et al J Biol Chem 299, 102893 (2023)
6. Asano et al Curr Protoc Cell Biol 80, e56 (2018)