Jul 27, 2023
Plasma procurement and processing
- hannah.anvari1,
- asma.giornazi1,
- shriya.shah1,
- maryellen.pavone1,
- francesca.e.duncan duncan1
- 1Northwestern Medicine OBGYN, Northwestern University
Protocol Citation: hannah.anvari, asma.giornazi, shriya.shah, maryellen.pavone, francesca.e.duncan duncan 2023. Plasma procurement and processing . protocols.io https://dx.doi.org/10.17504/protocols.io.14egn3z6ml5d/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 11, 2023
Last Modified: July 27, 2023
Protocol Integer ID: 84851
Keywords: human, blood, plasma
Funders Acknowledgement:
SenNet
Grant ID: U54AG075932
Abstract
Purpose: This protocol is intended for use in the collection and storage of human plasma in a research setting. The protocol details the collection, processing, and long-term storage for downstream analysis alongside other tissues or samples of interest (i.e. follicular fluid, ovary).
Guidelines
Researchers will adhere to all safety and training protocols required by the institution (Northwestern Medicine/Northwestern
University) including but not limited to:
- Biosafety Certification
- Bloodborne Pathogens Certification
- Collaborative Institutional TrainingInitiative (CITI program)Certification
Materials
- BD Vacutainer PlasticBlood Collection Tubes with K2EDTA: Tube Stopper (FisherScientific, BD 36866 or equivalent).
- Corning 2 mL InternalThreaded Polypropylene CryogenicVial, Self-Standing with Round Bottom(Fisher Scientific, 03-374-059 or equivalents).
Safety warnings
Researchers will wear personal protective equipment when working with human specimens, including gloves, masks, and lab coats.
Ethics statement
Human blood procurement and processing will adhere to the approved IRB protocol approved through NU (NU12G09) for collection of biofluids associated with study participants through Northwestern Medicine.
Blood samples in BD Vacutainer™ Plastic Blood Collection Tubes with K2EDTA with Tube Stopper (Fisher Scientific, 23-021-013) are collected in pre-op from nurses. Samples are kept upright and placed on ice immediately following collection. Research coordinator will bring both specimens to the lab on ice (2-4 °C). Samples should be transported in blue, labeled biohazard cooler bag (Fig. 1A, B).
Fig. 1. Typical blood/plasma collection. Whole blood samples are collected from clinical staff and then passed to research coordinator. Samples are brought to the lab on ice and kept at 2-4°C until processed (A, B).
Blood is centrifuged at 300g for 10min at room temperature (RT). Images of blood samples before and after centrifugation are seen below (Fig. 2A, B). Plasma (translucent top layer) is then aliquoted into 2 mL cryovial tubes (Fig. 2C) and stored at -80°C.
Fig. 2. Whole blood is centrifuged at 300g for 10 minutes (A). Once the plasma is clearly separated, the aspirate is collected and aliquoted into 2 mL cryovials (B, C). Samples are kept cold throughout processing (2-4°C) and then stored at -80°C.