On chip cryo-anesthesia of Drosophila larvae for high resolution in vivo imaging applications.
On chip cryo-anesthesia of Drosophila larvae for high resolution in vivo imaging applications.
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DOI：
10.1039/c7lc00345e
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发表时间：
2017-06-27
期刊：
影响因子：
6.1
通讯作者：
Chronis N
中科院分区：
文献类型：
--
作者：
Chaudhury AR;Insolera R;Hwang RD;Fridell YW;Collins C;Chronis N
我们提出了一种微流体芯片，用于固定果蝇幼虫以进行高分辨率体内成像。该芯片创造了一个低温微环境，可在 3 分钟内麻醉并固定幼虫。我们表征了芯片内的温度分布，并使用高分辨率荧光成像分析了由此产生的幼虫身体运动。我们的结果表明，所提出的方法最大限度地减少了内脏和组织的亚微米运动，而不影响幼虫的生理机能。它可用于短时间（几分钟）或较长时间（数小时）（如果间歇使用）连续固定幼虫。同一芯片可用于容纳和固定所有发育阶段（第一龄至第三龄晚期）的幼虫，并且将幼虫装载到芯片上不需要任何专门技能。为了证明芯片的可用性，我们观察了完整幼虫中神经元从细胞体到轴突末端的线粒体运输，以及线粒体融合和神经突触生长。除了研究亚细胞过程和细胞发育之外，我们还设想在各种生物体内成像应用中使用片上冷冻麻醉，包括观察唾液腺、脂肪体和体壁肌肉的器官发育。我们提出了一种用于冷冻固定果蝇幼虫的微流体芯片，用于神经生物学过程和其他应用的高分辨率体内成像。
We present a microfluidic chip for immobilizing Drosophila melanogaster larvae for high resolution, in vivo imaging. The chip creates a low-temperature micro-environment that anaesthetizes and immobilizes the larva in under 3 minutes. We characterized the temperature distribution within chip and analyzed the resulting larval body movement using high resolution fluorescence imaging. Our results indicate that the proposed method minimizes submicron movements of internal organs and tissue without affecting the larva physiology. It can be used to continuously immobilize larvae for short periods of time (minutes) or for longer periods (several hours) if used intermittingly. The same chip can be used to accommodate and immobilize larvae across all developmental stages (1st instar to late 3rd instar), and loading larvae onto the chip does not require any specialized skills. To demonstrate the usability of the chip, we observed mitochondrial trafficking in neurons from the cell bodies to the axon terminals along with mitochondrial fusion and neuro-synaptic growth through time in intact larvae. Besides studying sub-cellular processes and cellular development, we envision the use of on chip cryo-anesthesia in a wide variety of biological in vivo imaging applications, including observing organ development of the salivary glands, fat bodies and body-wall muscles. We present a microfluidic chip for cryo-immobilizing Drosophila melanogaster larvae for high resolution, in vivo imaging of neurobiological processes and other applications.
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影响因子：
7.4
作者：
Heo, Yun Seok;Cabrera, Lourdes M.;Takayama, Shuichi
通讯作者：
Takayama, Shuichi
DOI：
10.1016/j.cbpb.2005.01.019
发表时间：
2005-03-01
影响因子：
2.3
作者：
Badre, NH;Martin, ME;Cooper, RL
通讯作者：
Cooper, RL
影响因子：
4.6
作者：
Heemskerk, Idse;Lecuit, Thomas;LeGoff, Loic
通讯作者：
LeGoff, Loic
影响因子：
4.6
作者：
Koštál V;Korbelová J;Štětina T;Poupardin R;Colinet H;Zahradníčková H;Opekarová I;Moos M;Šimek P
通讯作者：
Šimek P
影响因子：
16.2
作者：
Gunawardena, S;Her, LS;Goldstein, LSB
通讯作者：
Goldstein, LSB