• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

使用胶原膜的活体成像友好型切片培养方法。

A live imaging-friendly slice culture method using collagen membranes.

机构信息

Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.

Isehara Research Laboratory, Technology and Development Division, Kanto Chemical Co., Inc, Suzukawa, Isehara, Kanagawa, Japan.

出版信息

Neuropsychopharmacol Rep. 2020 Sep;40(3):307-313. doi: 10.1002/npr2.12128. Epub 2020 Aug 5.

DOI:10.1002/npr2.12128
PMID:32757372
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7722644/
Abstract

AIM

Organotypic brain slice culture preserves the geographical position of neurons and neuronal circuits. The slice cultures also maintain both non-neuronal cell types and the surrounding extracellular matrix. The interface method has been widely used for slice cultures, in which brain slices are placed on semiporous polytetrafluoroethylene (PTFE) membranes. However, a low optical transparency of PTFE membrane makes it difficult to perform live imaging of deep regions of slice cultures using an inverted microscope. To overcome the issue, we evaluated the suitability of using collagen membranes for slice cultures, especially focusing on live imaging of the cellular dynamics of green fluorescent protein (GFP)-expressing microglia.

METHODS

Entorhinohippocampal slices were cultured on either collagen or PTFE membranes. The influence of membrane type on the ability to observe deep regions of slice cultures was examined by live imaging using an inverted microscope.

RESULTS

Collagen membranes were thinner and had better optical transparency compared with PTFE membranes. There were no differences in cell viability, density of neurons or microglia. The densify of visible short branches of microglia in live imaging was higher in collagen membranes than PTFE membranes.

CONCLUSION

Collagen membranes are suitable for live imaging of cellular dynamics in slice cultures using an inverted microscope.

摘要

目的

器官型脑片培养保留了神经元和神经元回路的地理位置。切片培养还维持了非神经元细胞类型和周围的细胞外基质。接口方法已广泛应用于切片培养，其中脑片置于半多孔聚四氟乙烯（PTFE）膜上。然而，PTFE 膜的低光学透明度使得使用倒置显微镜对切片培养的深部区域进行活细胞成像变得困难。为了克服这个问题，我们评估了胶原膜用于切片培养的适用性，特别是特别关注表达绿色荧光蛋白（GFP）的小胶质细胞的细胞动力学的活细胞成像。

方法

在胶原或 PTFE 膜上培养内嗅-海马脑片。通过使用倒置显微镜进行活细胞成像，检查膜类型对观察切片培养深部区域的能力的影响。

结果

胶原膜比 PTFE 膜更薄，光学透明度更好。细胞活力、神经元或小胶质细胞的密度没有差异。在活细胞成像中，胶原膜中小胶质细胞可见短分支的密集度高于 PTFE 膜。

结论

胶原膜适用于使用倒置显微镜对切片培养中的细胞动力学进行活细胞成像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/28a2961c049e/NPR2-40-307-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/068e0cd00f04/NPR2-40-307-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/a7987a9c05c2/NPR2-40-307-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/af8fa28f9ccb/NPR2-40-307-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/28a2961c049e/NPR2-40-307-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/068e0cd00f04/NPR2-40-307-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/a7987a9c05c2/NPR2-40-307-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/af8fa28f9ccb/NPR2-40-307-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cba/7722644/28a2961c049e/NPR2-40-307-g004.jpg

相似文献

1
A live imaging-friendly slice culture method using collagen membranes.使用胶原膜的活体成像友好型切片培养方法。
Neuropsychopharmacol Rep. 2020 Sep;40(3):307-313. doi: 10.1002/npr2.12128. Epub 2020 Aug 5.
2
Culturing thick brain slices: an interstitial 3D microperfusion system for enhanced viability.培养厚脑切片：一种用于提高存活率的间质三维微灌注系统。
J Neurosci Methods. 2009 Jun 15;180(2):243-54. doi: 10.1016/j.jneumeth.2009.03.016. Epub 2009 Mar 28.
3
Coupling of organotypic brain slice cultures to silicon-based arrays of electrodes.器官型脑片培养物与硅基电极阵列的耦合。
Methods. 1999 Jun;18(2):160-72. doi: 10.1006/meth.1999.0769.
4
Confocal imaging of microglial cell dynamics in hippocampal slice cultures.海马切片培养中小胶质细胞动态的共聚焦成像。
Methods. 1999 Jun;18(2):222-30, 177. doi: 10.1006/meth.1999.0775.
5
Preparation of Organotypic Slice Cultures for the Study of Glutamate Receptor Function.用于研究谷氨酸受体功能的器官型切片培养物的制备
Methods Mol Biol. 2019;1941:57-64. doi: 10.1007/978-1-4939-9077-1_5.
6
Preparation of Rat Organotypic Hippocampal Slice Cultures Using the Membrane-Interface Method.采用膜界面法制备大鼠器官型海马脑片培养物。
Methods Mol Biol. 2021;2188:243-257. doi: 10.1007/978-1-0716-0818-0_12.
7
The developmental expression of fluorescent proteins in organotypic hippocampal slice cultures from transgenic mice and its use in the determination of excitotoxic neurodegeneration.转基因小鼠海马脑片培养物中荧光蛋白的发育表达及其在兴奋性毒性神经退行性变测定中的应用。
Altern Lab Anim. 2007 Mar;35(1):61-70. doi: 10.1177/026119290703500121.
8
Characterization of cortical neuronal and glial alterations during culture of organotypic whole brain slices from neonatal and mature mice.原代培养新生和成年小鼠全脑脑片过程中皮质神经元和神经胶质改变的特征。
PLoS One. 2011;6(7):e22040. doi: 10.1371/journal.pone.0022040. Epub 2011 Jul 15.
9
Replenishment of Organotypic Hippocampal Slice Cultures with Neonatal or Adult Microglia.用新生或成年小胶质细胞补充器官型海马切片培养物。
Methods Mol Biol. 2019;2034:127-147. doi: 10.1007/978-1-4939-9658-2_10.
10
Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures.与器官型海马脑片培养物耦合的硅基电极阵列的生物相容性。
Brain Res. 2001 Mar 30;896(1-2):1-17. doi: 10.1016/s0006-8993(00)03304-7.

引用本文的文献

1
Ultra-thin and ultra-porous nanofiber networks as a basement-membrane mimic.超薄多孔纳米纤维网络作为基底膜模拟物。
Lab Chip. 2023 Oct 10;23(20):4565-4578. doi: 10.1039/d3lc00304c.
2
Replacement of Mouse Microglia With Human Induced Pluripotent Stem Cell (hiPSC)-Derived Microglia in Mouse Organotypic Slice Cultures.在小鼠器官型切片培养物中用人诱导多能干细胞（hiPSC）衍生的小胶质细胞替代小鼠小胶质细胞。
Front Cell Neurosci. 2022 Jul 13;16:918442. doi: 10.3389/fncel.2022.918442. eCollection 2022.
3
Assessing Microglial Dynamics by Live Imaging.

本文引用的文献

1
Visualization and Live Imaging of Oligodendrocyte Organelles in Organotypic Brain Slices Using Adeno-associated Virus and Confocal Microscopy.利用腺相关病毒和共聚焦显微镜对器官型脑片中少突胶质细胞细胞器进行可视化和实时成像
J Vis Exp. 2017 Oct 23(128):56237. doi: 10.3791/56237.
2
Depth and time-dependent heterogeneity of microglia in mouse hippocampal slice cultures.小鼠海马切片培养物中微胶质细胞的深度和时间依赖性异质性
Neurosci Res. 2016 Oct;111:64-9. doi: 10.1016/j.neures.2016.05.001. Epub 2016 May 17.
3
A simple method for multiday imaging of slice cultures.
通过实时成像评估小胶质细胞动力学。
Front Immunol. 2021 Mar 8;12:617564. doi: 10.3389/fimmu.2021.617564. eCollection 2021.
一种用于切片培养多日成像的简单方法。
Microsc Res Tech. 2010 Jan;73(1):37-44. doi: 10.1002/jemt.20750.
4
Bone regeneration using collagen type I vitrigel with bone morphogenetic protein-2.使用含骨形态发生蛋白-2的I型胶原蛋白玻璃凝胶进行骨再生。
J Biosci Bioeng. 2009 Mar;107(3):318-23. doi: 10.1016/j.jbiosc.2008.10.007.
5
Regressive events in rat corticospinal axons during development in in vitro slice cocultures: retraction, amputation, and degeneration.体外切片共培养中大鼠皮质脊髓轴突发育过程中的退行性事件：回缩、截断和变性。
J Comp Neurol. 2009 Mar 10;513(2):164-72. doi: 10.1002/cne.21950.
6
A low-cost method for brain slice cultures.一种用于脑片培养的低成本方法。
J Pharmacol Sci. 2007 Jun;104(2):191-4. doi: 10.1254/jphs.sc0070119. Epub 2007 Jun 8.
7
The growth of cells on a transparent gel of reconstituted rat-tail collagen.细胞在重组大鼠尾胶原透明凝胶上的生长。
J Natl Cancer Inst. 1956 Jun;16(6):1375-403.
8
Organotypic slice cultures: a technique has come of age.器官型切片培养：一项技术已走向成熟。
Trends Neurosci. 1997 Oct;20(10):471-7. doi: 10.1016/s0166-2236(97)01122-3.
9
Stop and branch behaviors of geniculocortical axons: a time-lapse study in organotypic cocultures.膝状皮质轴突的终止和分支行为：器官型共培养的延时研究
J Neurosci. 1997 May 15;17(10):3653-63. doi: 10.1523/JNEUROSCI.17-10-03653.1997.
10
Dynamics of target recognition by interstitial axon branching along developing cortical axons.沿发育中的皮质轴突的间质轴突分支进行目标识别的动力学
J Neurosci. 1996 Feb 15;16(4):1450-9. doi: 10.1523/JNEUROSCI.16-04-01450.1996.