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    Plasma procurement and processing

    hannah.anvari, asma.giornazi, shriya.shah, maryellen.pavone, francesca.e.duncan duncan

    Published: 2023-07-27 DOI: 10.17504/protocols.io.14egn3z6ml5d/v1
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    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).

    Steps

    1.

    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).
    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).
    2.

    Blood is centrifuged at 300 g 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.
    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.

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