流式检测新鲜和冻存痰液标本中的白细胞

niwanmao

此进展可能对呼吸科疾病更有帮助。本文作者利用流式结合CD45、CD14、CD16等抗体检测了新鲜和冻存痰液标本中的白细胞分类，并与传统光镜下的细胞分类计数对比，发现两者一致，而流式较传统光镜下分类更稳定、更客观。经过－80℃冻存后，粒细胞和单核细胞容易受到破坏，淋巴细胞相对更稳定。

摘要如下：

Cytometry B Clin Cytom. 2013 Jan 22. doi: 10.1002/cyto.b.21069. [Epub ahead of print]  Identifying leukocyte populations in fresh and cryopreserved sputum using flow cytometry.Brooks CR   , van Dalen CJ   , Hermans IF   , Douwes J   .SourceCentre for Public Health Research, Massey University Wellington Campus, Wellington, New Zealand. c.r.brooks@massey.ac.nz.
AbstractBACKGROUND:Airway inflammation is commonly assessed by sputum induction followed by a differential cell count (DCC) using light microscopy. This method is prone to intercounter variability and poor reproducibility. We aimed to develop a more objective method using flow cytometry (FCM).METHODS:Fifty-six sputum inductions were conducted in 41 adults (23 asthmatics). Sputum was processed, a cytospin prepared for DCC, and the remainder immunolabeled for FCM using CD45, CD14, and CD16-specific antibodies to distinguish major leukocyte populations. Aliquots of 15 samples were frozen at -80°C to assess the effects of cryostorage. DCC and FCM were compared, and viability of individual cell populations was determined by FCM.RESULTS:FCM and DCC, and fresh and frozen samples, were significantly correlated, R = 0.54-0.87; all P < 0.0001, and R = 0.57 to 1; P < 0.005, respectively. There was a significant neutrophil loss after cryostorage (from median 30.5-17.4% of total leukocytes; P < 0.0001). Cell viability was higher for lymphocytes compared to granulocytes or macrophages (P < 0.001). With the exception of the expected higher levels of eosinophils (P < 0.005), no significant difference in cell differentials or viability was observed between asthmatics and nonasthmatics using either DCC or FCM.CONCLUSIONS:FCM is a suitable means of assessing leukocyte populations in induced sputum. Sample storage at -80°C prior to FCM is feasible, but may be detrimental to neutrophils, although good correlations were still observed between fresh and frozen samples. Large differences in viability were found between individual cell populations suggesting that viability dye use may be necessary. © 2013 International Clinical Cytometry Society.PMID: 23341171

全文链接：http://dx.doi.org/10.1002/cyto.b.21069

culturelily

看来还是得现做现测