Optimized protocol for isolation of neutrophils from small-volume human blood samples via Percoll density gradient centrifugation
- 1Department of Rheumatology & Immunology, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Children and Adolescents’ Health and Diseases, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Child Rare Diseases in Infection and Immunity, Chongqing 400014, China
Abstract
Neutrophils are critical first-line innate immune defenders. Conventional Percoll-based isolation protocols perform reliably only with large blood volumes (e.g., 10 mL) and yield inconsistent results from small-volume samples (≤2 mL), such as those from pediatric individuals. Here, we present an optimized protocol tailored for minimal-volume specimens, with systematic adjustments to centrifugation speed, duration, and gradient setup to ensure neutrophil isolation with high purity and viability. Among healthy donor samples (n = 19), the protocol yielded 1.74 ± 0.57 × 106 granulocytes per mL of blood. Across all tested samples (n = 22), granulocyte purity was 91.58% ± 3.70%, and neutrophils comprised 96.48% ± 1.75% of recovered granulocytes in six evaluated isolates. In comparison, conventional protocols yielded granulocytes with low purity of 33.25% ± 13.99%. This robust, reproducible method enables reliable neutrophil isolation, expanding the accessibility of neutrophil-related research to scenarios with limited blood volume.
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BibTeX
@article{she2026optimized,
title = {{Optimized protocol for isolation of neutrophils from small-volume human blood samples via Percoll density gradient centrifugation}},
author = {Rui She and Shaotian Chen and Zhiquan Liu and Xuemei Tang and Xiang Qiu and Weijian Chen},
journal = {STAR Protocols},
year = {2026},
month = oct,
doi = {10.1016/j.xpro.2026.104914},
url = {https://doi.org/10.1016/j.xpro.2026.104914},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Optimized protocol for isolation of neutrophils from small-volume human blood samples via Percoll density gradient centrifugation
AU - Rui She
AU - Shaotian Chen
AU - Zhiquan Liu
AU - Xuemei Tang
AU - Xiang Qiu
AU - Weijian Chen
PY - 2026
DA - 2026/10/28
JO - STAR Protocols
DO - 10.1016/j.xpro.2026.104914
UR - https://doi.org/10.1016/j.xpro.2026.104914
AB - Neutrophils are critical first-line innate immune defenders. Conventional Percoll-based isolation protocols perform reliably only with large blood volumes (e.g., 10 mL) and yield inconsistent results from small-volume samples (≤2 mL), such as those from pediatric individuals. Here, we present an optimized protocol tailored for minimal-volume specimens, with systematic adjustments to centrifugation speed, duration, and gradient setup to ensure neutrophil isolation with high purity and viability. Among healthy donor samples (n = 19), the protocol yielded 1.74 ± 0.57 × 106 granulocytes per mL of blood. Across all tested samples (n = 22), granulocyte purity was 91.58% ± 3.70%, and neutrophils comprised 96.48% ± 1.75% of recovered granulocytes in six evaluated isolates. In comparison, conventional protocols yielded granulocytes with low purity of 33.25% ± 13.99%. This robust, reproducible method enables reliable neutrophil isolation, expanding the accessibility of neutrophil-related research to scenarios with limited blood volume.
SN - 2666-1667
ER -