Blood collection technique, anticoagulants and storing temperature have minor effects on the isolation of polymorphonuclear neutrophils
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Updated: Dec 10, 2025
Isolation of Human Neutrophils from Whole Blood and Buffy Coats
Published on: September 17, 2021
Julia Krabbe1, Viktor Beilmann2,3, Hanif Alamzad-Krabbe3
1Institute of Occupational, Social and Environmental Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany. jkrabbe@ukaachen.de.
Abstract:
In the isolation of polymorphonuclear neutrophils (PMNs) the technique and other external factors can have great influence on the quality and quantity of isolated neutrophils. To elucidate the influence of the blood collection technique, anticoagulants and storing temperature on isolated PMNs healthy volunteers provided blood samples with different needles and collection techniques, anticoagulants (EDTA, heparin, citrate) and storing temperatures (4, 22, 37 °C). From each blood sample PMNs were isolated and compared regarding number of PMNs and oxidative burst. The blood collection technique, anticoagulants and storing temperature had minor impact on isolated PMNs. All three tested cannulas and anticoagulants can be used to obtain blood samples for PMN isolation. For storing temperatures 37 °C should be preferred. Regarding time between the PMN isolation and the actual experiments, a time span of maximum 1 h should be targeted.
Insights
Blood collection techniques, anticoagulants, and storage temperatures have minimal impact on isolating polymorphonuclear neutrophils (PMNs). Optimal PMN isolation requires prompt experimentation within one hour post-collection, with 37°C preferred for sample storage.
Area of Science:
- Immunology
- Hematology
Background:
- Optimizing the isolation of polymorphonuclear neutrophils (PMNs) is crucial for accurate experimental results.
- External factors during blood collection and processing can significantly affect PMN yield and function.
Purpose of the Study:
- To investigate the impact of blood collection techniques, anticoagulants, and storage temperatures on the quality and quantity of isolated PMNs.
- To establish best practices for PMN isolation to ensure reliable experimental outcomes.
Main Methods:
- Blood samples were collected from healthy volunteers using various needles and collection methods.
- Different anticoagulants (EDTA, heparin, citrate) and storage temperatures (4°C, 22°C, 37°C) were tested.
- Isolated PMNs were analyzed for number and oxidative burst activity.
Main Results:
- Blood collection technique and anticoagulants showed minor effects on PMN isolation.
- Storage temperature had a minor impact, with 37°C being the preferred condition.
- A maximum time span of one hour between PMN isolation and experimentation is recommended.
Conclusions:
- Standard blood collection techniques and common anticoagulants are suitable for PMN isolation.
- Storing blood samples at 37°C is recommended for optimal PMN isolation.
- Minimizing the time between blood collection and experimentation to one hour is critical for preserving PMN integrity and function.
