Eliminating Freeze-thaw Cycles: Advances in Frozen Biospecimen Integrity Utilization

Quick and effective frozen sample processing for applications in fields such as the newly expanding area of molecular epidemiology, or the more classic fields of molecular biology and toxicology, has always been a dream for investigators in these and other research areas. Repeated freeze-thaw cycles are detrimental to biospecimen quality and often result in using up the whole sample in a single shot, frequently resulting in either unnecessary waste of biological material or limitations to the breadth of investigations that can be conducted. While such specific applications in practical lab operations and management are often overlooked by the mainstream scientific news, this topic always ends up standing in the way of a cost-effective and high quality analysis of frozen biological samples.

This webinar is intended to provide the scientific community with information on new approaches for quick and effective frozen sample processing for experimental applications by:

• Summarizing the challenges of a cost-effective and high quality analysis of frozen biological samples
• Looking to strategies for protecting sample quality from cycles of harmful repeated thawing and refreezing by using the CryoXtract frozen aliquotting technology
• Analyzing the impact of CryoXtract frozen aliquotting technology on expanding the information that can be extracted from biological samples
• Designing a pipeline for sample processing that may be specifically applied to the molecular epidemiology and toxicology fields

Each and every scientist in fields such as molecular biology, molecular epidemiology and toxicology, spends a lot of time and resources generating/collecting and storing precious tissue and cell samples. Each and every scientist also knows that probably the best way to save precious samples is by freezing them and hosting them in refrigerated units with temperature ranges spanning from the more standard -20°C freezer all the way to liquid nitrogen cells (-196°C).

However, there can be a great deal of uncertainty when it comes time to either plan how to store and, later on, how to make use of such precious samples. Some scientists for whom space is not an issue (and thanks to ever-developing sample barcoding techniques), go for aliquotting samples in multiple vials. This approach allows for keeping a lasting amount of tissue frozen for the conduction of studies that were probably not even possible at the time the samples were collected. On the other hand, this approach leads to extensive collections of vials that have a big footprint and carries with it high maintenance costs. Other scientists instead orient themselves toward the generation of individual aliquots, often due to space and cost issues. However, they pay the price when it comes time to analyze their samples because they either have to settle for “one-study-takes-all” or, in the best case scenario, deal with a decrease in sample quality because thawing and refreezing cycles limit the amount of scientific data that can be extracted from the specimen. While the first scenario is available to few researchers, it poses the typical question of “how big the aliquot?” The second scenario is much more common.

In this webinar the presenter will discuss the application of a third possible scenario that, while limiting the cost, allows use of tissue for multiple applications while simultaneously preserving the quality of the sample in its original status. The CryoXtract frozen aliquotter instrument allows for drilling through frozen biopsies to retrieve portions of the tissue of variable sizes by using a hollow bit installed on a drilling head that drills through the host sample, within its storage vial, by a liquid nitrogen-cooled (~-196°C) holder.

The lab has made extensive use of CryoXtract’s instrument. As a lab devoted to the molecular epidemiology field, it often deals with human sample cohorts of several hundred collected with a great deal of effort in the field and the investment of substantial financial resources. The optimization of the use of such samples by enabling the extraction of the maximum amount of information possible in order to generate integrated, and therefore highly informative, datasets is critical.

In CryoXtract’s experience, the use of CryoXtract frozen aliquotting technology has allowed it, specifically, to repeatedly “sample” their specimens, without the need to expose them to freeze-thaw cycles, to extract high quality DNA and RNA that has been used for many different techniques spanning from the more basic standard and real-time PCR to the more advanced microarray and sequencing applications. The CryoXtract frozen aliquotting technology also perfectly fits into its carefully designed pipeline that allows a significant reduction of the time needed to go from frozen tissue samples to DNA/RNA. At this time, considering specimens’ sampling by using the CryoXtract frozen aliquotting technology, tissue homogenization and DNA/RNA extraction, technicians can safely process approximately 100 samples in a 2-week time period.

Speaker

Luca Lambertini, PhD, MSc, MPH,Assistant Professor of Preventive Medicine and Obstetrics, Gynecology and Reproductive Science (OB/GYN), Icahn School of Medicine at Mount Sinai

Luca Lambertini is a member of the Senator Frank R. Lautenberg Environmental Health Sciences Laboratory, a state-of-the-art laboratory equipped with the latest technology and instruments to conduct cutting-edge research in environmental health at the Icahn School of Medicine at Mount Sinai (ISMMS) Preventive Medicine Department. He works within the molecular epidemiology unit of the lab where he focuses his activity on the identification and characterization of placental biomarkers of the effects of the exposure to different environmental stimuli on alterations of the fetoplacental development. Luca’s work is primarily devoted to the study of nuclear and mitochondrial placenta epi/genetics in different cohort studies.

During his 9-year activity at ISMMS, Luca also developed, implemented and directed the Mount Sinai Pregnancy Biobank, a tissue bank oriented at collecting and store placenta and umbilical cord blood samples from pregnant women that deliver at the ISMMS OB/GYN Department for research applications.

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