|
发表于 2013-10-15 06:47:40
|
显示全部楼层
niwanmao 发表于 2013-10-14 20:37
我怎么在这篇文章里面没看到相关的技术介绍嘛,只看到呼吸量测定、组织学、细胞因子等测定。 ...
里面有一小段提到这个chipcytometry技术的使用,在902页的右侧中间的cytokine and chemokine detection里面有“To investigate the cellular source of the measured cytokines in BAL fluid, multi-marker chipcytometry was applied to two additional
mice per time point at 0, 4, 24, 72 h post inoculation (Hennig
et al. 2009; www.chipcytometry.com). 10 l cell-containing BAL
fluid were pipetted into the microfluidic-chip without prior ex
vivo modification. Cells were fixed inside the chip using 2%
paraformaldehyde (PFA) and afterwards analyzed with a comprehensive
marker set (clone, company and dilution in brackets):
CD11c [HL3, BD, 1:20], Gr-1 [RB6-8C5, BD, 1:400], F4/80 [BM8,
eBiosciences, 1:100], CD68 [FA-11, Acris, 1:20], TNF-alpha [MP6-
XT22, R&D, 1:50], IL-10 [JES5-16E3, BD, 1:50], IFN-gamma [XMG1.2,
BD, 1:150], IL-2 [JES6-5H4, eBiosciences, 1:20], IL-4 [11B11, BD,
1:20], IL-5 [TRFK5, BD, 1:20], IL-6 [MP5-20F3, BD, 1:20], IL17A
[eBio17B7, eBioscience, 1:20]). All antibodies were used phycoerythrin
(PE)-labeled, incubation times at room temperature per
antibody: 5 min in PBS for surface markers, 1 h in PermWash-Buffer
(BD Biosciences) for intracellular markers.” |
|