Teasing Out Fine Details from the Tangled World of Chromosomes
The Stumpff Lab, Department of Molecular Physiology and Biophysics, University of Vermont
Normal human cells contain 23 chromosome pairs, each encoding the genetic information of life as well as passing on inheritable traits from parents. Chromosomes are tightly folded into origami-like structures that are mind-boggling in their complexity, yet elegantly unfurled and refolded during cell division or mitosis. Tiny errors during mitosis can cause the formation of aneuploid cells containing an abnormal number of chromosomes and lead to miscarriage and health disorders such as Down syndrome. Additionally, these mistakes have been linked to tumorigenesis and accelerated tumor growth rates.
At the University of Vermont’s Department of Molecular Physiology and Biophysics, Dr. Jason Stumpff’s lab is particularly interested in microtubule dynamics and cell division, and how these mechanisms can be leveraged to preserve genomic integrity for normally functioning cells, or conversely, to damage or destroy abnormally functioning tumor cells. Most recently, the Stumpff lab was taking a close look at a particular molecule used in cell division as a target to inhibit triple negative breast cancer. This particularly rare and aggressive cancer lacks receptors for estrogen, progesterone and HER-2, thus rendering common treatments ineffective. The assays used with the cancer cells and other research in the lab included live cell imaging and other advanced microscopy techniques during the cell division process to visualize the connections between chromosomes and the molecular machinery that segregates them, and then how that machine is regulated.
Existing manual imaging systems in the facility were limited in throughput and weren’t sensitive enough to detect subtle changes. As Dr. Stumpff discussed his experimental intent and existing imaging limitations with BioTek’s applications scientists, he learned about BioTek’s BioSpa™ Live Cell Analysis System, consisting of the Cytation™ 5 Cell Imaging Multi-Mode Reader and BioSpa 8 Automated Incubator. BioTek’s imaging experts worked with the Stumpff lab to design a sensitive kinetic proliferation assay in 96-well format for use in the BioSpa System.
In the hands-free assay, microplates containing cells are incubated using optimal environmental conditions in the BioSpa 8, then automatically transferred to the Cytation 5 for high contrast brightfield imaging every two hours over four days. Gen5™ Software automatically counts the cells and creates reproducible growth curves, and can also create time-lapse movies of each well. “The BioSpa System adds a new dimension to the lab’s research capabilities and reduces the time-consuming bottleneck of manually analyzing images to produce quantitative data,” Dr. Stumpff comments. Additionally, he cites the enhanced throughput, variety of imaging modes, parallel imaging of different cell treatments, long-term imaging and built-in analysis tools as being particularly beneficial in the course of his research.
The lab is also currently working with BioTek’s Applications Group to develop an assay to screen for CRISPR-modified cell lines and continues to seek ways to enhance their research through the use of the BioSpa System. Dr. Stumpff is quick to note that it’s not just BioTek’s products that deliver beyond expectations. The swift response to service requests minimizes any potential system downtime, and multiple BioTek teams consistently strive to ensure that the BioSpa System is as useful as possible for the Stumpff lab’s specific research questions and goals. He summarizes his experience, saying, “I’m really impressed with the willingness of BioTek’s applications and engineering teams to invest time with us, to help us fine-tune our assays and to respond to our needs with new insights and tools. It’s been a huge plus, and we appreciate that they go the extra mile for us.”
Colorectal cancer cells imaged in high contrast brightfield using the Cytation 5 system (grayscale)
and auto-counted with Gen5 software (yellow outlines).
To learn more about The Stumpff Lab in the University of Vermont’s Department of Molecular Physiology and Biophysics, visit their web site.
Thanks to Dr. Jason Stumpff for sharing his BioTek experience.