IR-Live: fabrication of a low-cost plastic microfluidic device for infrared spectromicroscopy of living cells
IR-Live: fabrication of a low-cost plastic microfluidic device for infrared spectromicroscopy of living cells
复制标题
DOI：
10.1039/c5lc01460c
复制
发表时间：
2016-01-01
期刊：
影响因子：
6.1
通讯作者：
Grenci, G.
中科院分区：
文献类型：
--
作者：
Birarda, G.;Ravasio, A.;Grenci, G.
水是一种强的中红外吸收剂，这阻碍了充分利用无标记和非侵入性红外（IR）光谱技术研究活体生物样品。为了克服这一障碍，许多研究人员已经建立了复杂的流体腔室或微流控芯片，其中的液体介质在样品隔室的深度被限制在10 μ m或less. Here，我们报告了一种创新的和简单的方法来制造塑料装置与红外透明的观察口，使活的生物样品的红外光谱显微镜，因此该设备被命名为“IR-Live”。这种方法的优点包括较低的生产成本，最小限度地需要访问一个微型制造设施，和无限的质量或废物交换的活样品周围的视口区域。我们证明，低成本的IR-Live与微流体灌注技术相结合，可以实现长期（>60小时）的细胞培养，这扩大了红外光谱显微镜研究活体生物样品的能力。为了说明这一点，我们首先应用该设备研究蛋白质和脂质极性迁移REF 52成纤维细胞通过收集2维光谱化学地图在微米空间分辨率。然后，我们证明了我们的方法来研究动态细胞事件的适用性，通过收集U937单核细胞在细胞附着到生物相容性表面的早期阶段的光谱图的时间序列。
Water is a strong mid-infrared absorber, which has hindered the full exploitation of label-free and noninvasive infrared (IR) spectromicroscopy techniques for the study of living biological samples. To overcome this barrier, many researchers have built sophisticated fluidic chambers or microfluidic chips wherein the depth of the liquid medium in the sample compartment is limited to 10 mu m or less. Here we report an innovative and simple way to fabricate plastic devices with infrared transparent view-ports enabling infrared spectromicroscopy of living biological samples; therefore the device is named "IR-Live". Advantages of this approach include lower production costs, a minimal need to access a micro-fabrication facility, and unlimited mass or waste exchange for the living samples surrounding the view-port area. We demonstrate that the low-cost IR-Live in combination with microfluidic perfusion techniques enables long term (>60 h) cell culture, which broadens the capability of IR spectromicroscopy for studying living biological samples. To illustrate this, we first applied the device to study protein and lipid polarity in migrating REF52 fibroblasts by collecting 2-dimensional spectral chemical maps at a micrometer spatial resolution. Then, we demonstrated the suitability of our approach to study dynamic cellular events by collecting a time series of spectral maps of U937 monocytes during the early stage of cell attachment to a bio-compatible surface.