Whole-blood fixation provides a rapid and simplified method for cell preservation compared to isolation of peripheral blood mononuclear cells (PBMCs). This can be especially important for sample acquisition and storage in resource-limited settings. However, some caveats have been reported, such as reduced cell marker recognition. Here, we evaluated the whole-blood proteomic stabilizer PROT1 and compared recognition of 53 common cell markers in fixed buffy coats and cryopreserved PBMCs isolated from the same donor. Several antibodies completely lost their binding to the cells, while others presented with partial loss of marker recognition or no effect at all. Based on the screened antibodies, we designed two antibody panels allowing phenotyping of B cells, monocytes, and dendritic cells and also T cells and NK cells in both fixed and non-fixed material. Taken together, our observations suggest that antibodies intended to be used with fixed blood first need to be evaluated for marker recognition and staining intensity, in comparison with fresh samples or cryopreserved PBMCs.

与分离外周血单核细胞（PBMC）相比，全血固定提供了一种快速，简化的细胞保存方法。这对于在资源有限的设置中进行样品采集和存储尤其重要。但是，已经报告了一些警告，例如减少的细胞标记识别。在这里，我们评估了全血蛋白质组稳定剂PROT1，并比较了从相同供体分离的固定血沉棕黄层和冷冻保存的PBMC中53种常见细胞标记的识别。几种抗体完全失去了与细胞的结合，而其他抗体则部分失去了标记识别或完全没有作用。根据筛选出的抗体，我们设计了两个抗体板，可以对B细胞，单核细胞，固定和非固定材料中的树突状细胞以及T细胞和NK细胞。综上所述，我们的观察结果表明，与新鲜样品或冷冻保存的PBMC相比，与固定血液一起使用的抗体首先需要进行标志物识别和染色强度评估。