A microfluidic platform for lifelong high-resolution and high throughput imaging of subtle aging phenotypes in C. elegans†,Lab on a Chip

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A microfluidic platform for lifelong high-resolution and high throughput imaging of subtle aging phenotypes in C. elegans†
Lab on a Chip ( IF 6.1 ) Pub Date : 2018-08-30 00:00:00 , DOI: 10.1039/c8lc00655e
Sahand Saberi-Bosari ₁ , Javier Huayta , Adriana San-Miguel

Affiliation

Aging produces a number of changes in the neuronal structure and function throughout a variety of organisms. These aging-induced changes encompass a wide range of phenotypes, from loss of locomotion ability to defective production of synaptic vesicles. C. elegans is one of the primary systems used to elucidate phenotypes associated with aging processes. Conventional aging studies in C. elegans are typically labor-intensive, low-throughput, and incorporate fluorodeoxyuridine (FUdR) as a sterilizing agent to keep the population age-synchronized throughout the assay. However, FUdR exposure induces lifespan extension, and can potentially mask the phenotypes associated with the natural aging process. In addition, studying cellular or subcellular structures requires anesthetics or adhesives to immobilize nematodes while acquiring high-resolution images. In this platform, we are able to maintain a population (∼1000 worms) age-synchronized throughout its lifespan and perform a series of high-resolution microscopy studies in a drug-free environment. The device is composed of two main interconnected sections, one with the purpose of filtering progeny while keeping the parent population intact, and one for trapping nematodes in individual compartments for microscopy. Immobilization is carried out by decreasing the temperature of the device where nematodes are trapped by placing a heat sink on top of the chip. We were able to perform periodic high-resolution microscopy of fluorescently tagged synapses located at the dorsal side of the nematode's tail throughout the worms' lifespan. To characterize the subtle phenotypes that emerge as nematodes age, computer vision was implemented to perform automated unbiased detection of synapses and quantitative analysis of aging-induced synaptic changes.

中文翻译：

一种微流体平台，可对线虫的微妙衰老表型进行终生高分辨率和高通量成像†

衰老会导致多种生物体的神经元结构和功能发生许多变化。这些衰老引起的变化涵盖了广泛的表型，从运动能力丧失到突触小泡产生缺陷。线虫是用于阐明与衰老过程相关的表型的主要系统之一。线虫的传统衰老研究通常是劳动密集型、低通量的，并采用氟脱氧尿苷 (FUdR) 作为灭菌剂，以在整个测定过程中保持种群年龄同步。然而，暴露于 FUdR 会导致寿命延长，并可能掩盖与自然衰老过程相关的表型。此外，研究细胞或亚细胞结构需要麻醉剂或粘合剂来固定线虫，同时获取高分辨率图像。在这个平台中，我们能够在整个生命周期中维持年龄同步的种群（约 1000 条线虫），并在无药物的环境中进行一系列高分辨率显微镜研究。该装置由两个主要的互连部分组成，一个的目的是过滤后代，同时保持亲代群体完整，另一个用于将线虫捕获在单独的隔间中进行显微镜检查。通过在芯片顶部放置散热器来降低捕获线虫的装置的温度来进行固定。在线虫的整个生命周期中，我们能够定期对位于线虫尾部背侧的荧光标记突触进行高分辨率显微镜观察。为了表征线虫衰老时出现的微妙表型，利用计算机视觉对突触进行自动无偏检测，并对衰老引起的突触变化进行定量分析。

更新日期：2018-08-30

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