Application of Light-Sheet Mesoscopy to Image Host-Pathogen Interactions in Intact Organs.
Application of Light-Sheet Mesoscopy to Image Host-Pathogen Interactions in Intact Organs.
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DOI：
10.3389/fcimb.2022.903957
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发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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非洲人类锥虫病（HAT）是一种由细胞外寄生虫布鲁氏锥虫（Trypanosoma bruei）引起的疾病，在感染的慢性阶段影响中枢神经系统（CNS），如不及时治疗，可引起神经炎症、昏迷和死亡。然而，人们对感染导致的大脑结构变化知之甚少。到目前为止，感染引起的神经炎症已经用传统的方法观察到，如免疫组织化学、电子显微镜和双光子显微镜，只在大脑的一小部分，这可能不是疾病的代表。在本文中，我们使用了一种新开发的光片照射器来成像慢性感染小鼠的神经炎症水平，并将其与naïve对照组进行比较。该系统是为与Mesolens物镜结合成像而开发的，可为数十毫米大的成像体积提供快速的亚细胞分辨率。用CUBIC+清除小鼠脑标本，然后用抗体染色定位表达胶质纤维酸蛋白（GFAP）的细胞，主要是星形胶质细胞和室管膜细胞，在这里用作细胞反应性和胶质细胞形成的代理。大的捕获量使我们能够检测GFAP+细胞并在空间上解决对布氏体感染的反应。基于GFAP+细胞的形态计量学分析和空间分布，我们的数据显示，侧脑室周围以及第三脑室背侧和腹侧的细胞树突分支显著增加，这与从心室周围空间向远端延伸的分支呈负相关。据我们所知，这是第一份强调光片肠系镜在完整清除器官细胞水平上表征小鼠大脑对寄生虫感染的炎症反应的潜力的报告，为开发用于研究宿主-病原体相互作用的新型中尺度成像技术开辟了新的途径。
Human African Trypanosomiasis (HAT) is a disease caused by the extracellular parasite Trypanosoma brucei that affects the central nervous system (CNS) during the chronic stage of the infection, inducing neuroinflammation, coma, and death if left untreated. However, little is known about the structural change happening in the brain as result of the infection. So far, infection-induced neuroinflammation has been observed with conventional methods, such as immunohistochemistry, electron microscopy, and 2-photon microscopy only in small portions of the brain, which may not be representative of the disease. In this paper, we have used a newly-developed light-sheet illuminator to image the level of neuroinflammation in chronically infected mice and compared it to naïve controls. This system was developed for imaging in combination with the Mesolens objective lens, providing fast sub-cellular resolution for tens of mm3-large imaging volumes. The mouse brain specimens were cleared using CUBIC+, followed by antibody staining to locate Glial Fibrillary Acid Protein (GFAP) expressing cells, primarily astrocytes and ependymocytes, used here as a proxy for cell reactivity and gliosis. The large capture volume allowed us to detect GFAP+ cells and spatially resolve the response to T. brucei infection. Based on morphometric analyses and spatial distribution of GFAP+ cells, our data demonstrates a significant increase in cell dendrite branching around the lateral ventricle, as well as dorsal and ventral third ventricles, that are negatively correlated with the branch extension in distal sites from the circumventricular spaces. To our knowledge, this is the first report highlighting the potential of light-sheet mesoscopy to characterise the inflammatory responses of the mouse brain to parasitic infection at the cellular level in intact cleared organs, opening new avenues for the development of new mesoscale imaging techniques for the study of host-pathogen interactions.
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影响因子：
3.8
作者：
Laperchia, Claudia;Palomba, Maria;Bentivoglio, Marina
通讯作者：
Bentivoglio, Marina
影响因子：
2.1
作者：
HERBERT, WJ;LUMSDEN, WHR
通讯作者：
LUMSDEN, WHR
影响因子：
8.8
作者：
Niedworok, Christian J.;Schwarz, Inna;Schwarz, Martin K.
通讯作者：
Schwarz, Martin K.
DOI：
10.1126/science.1257998
发表时间：
2014-10-24
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen BC;Legant WR;Wang K;Shao L;Milkie DE;Davidson MW;Janetopoulos C;Wu XS;Hammer JA 3rd;Liu Z;English BP;Mimori-Kiyosue Y;Romero DP;Ritter AT;Lippincott-Schwartz J;Fritz-Laylin L;Mullins RD;Mitchell DM;Bembenek JN;Reymann AC;Böhme R;Grill SW;Wang JT;Seydoux G;Tulu US;Kiehart DP;Betzig E
通讯作者：
Betzig E
影响因子：
48
作者：
Planchon, Thomas A.;Gao, Liang;Milkie, Daniel E.;Davidson, Michael W.;Galbraith, James A.;Galbraith, Catherine G.;Betzig, Eric
通讯作者：
Betzig, Eric