Imaging of human differentiated 3D neural aggregates using light sheet fluorescence microscopy

被引:32
作者
Gualda, Emilio J. [1 ]
Simao, Daniel [2 ,3 ]
Pinto, Catarina [2 ,3 ]
Alves, Paula M. [2 ,3 ]
Brito, Catarina [2 ,3 ]
机构
[1] Inst Gulbenkian Ciencias, Cell Imaging Unit, P-2780156 Oeiras, Portugal
[2] Inst Biol Expt & Tecnol, Adv Cell Models Lab, Oeiras, Portugal
[3] Univ Nova Lisboa, Inst Tecnol Quim & Biol, Anim Cell Technol Unit, Oeiras, Portugal
来源
FRONTIERS IN CELLULAR NEUROSCIENCE | 2014年 / 8卷
关键词
fluorescence microscopy; light-sheet fluorescence microscopy; 3D neural aggregates; calcium imaging; live/dead assays; PLANE ILLUMINATION MICROSCOPY; PLURIPOTENT STEM-CELLS; CULTURE; DEEP;
D O I
10.3389/fncel.2014.00221
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The development of three dimensional (3D) cell cultures represents a big step for the better understanding of cell behavior and disease in a more natural like environment, providing not only single but multiple cell type interactions in a complex 3D matrix, highly resembling physiological conditions. Light sheet fluorescence microscopy (LSFM) is be coming an excellent tool for fast imaging of such 3D biological structures. We demonstrate the potential of this technique for the imaging of human differentiated 3D neural aggregates in fixed and live samples, namely calcium imaging and cell death processes, showing the power of imaging modality compared with traditional microscopy. The combination of light sheet microscopy and 3D neural cultures will open the door to more challenging experiments involving drug testing at large scale as well as a better understanding of relevant biological processes in a more realistic environment.
引用
收藏
页数:10
相关论文
共 39 条
[1]  
Ansari N., 2013, FOCUS MICROSCOPY
[2]   Gene Therapy for Parkinson's Disease [J].
Bjorklund, Tomas ;
Kordower, Jeffrey H. .
MOVEMENT DISORDERS, 2010, 25 (03) :S161-S173
[3]   Three-dimensional cell culture: the missing link in drug discovery [J].
Breslin, Susan ;
O'Driscoll, Lorraine .
DRUG DISCOVERY TODAY, 2013, 18 (5-6) :240-249
[4]   Generation and genetic modification of 3D cultures of human dopaminergic neurons derived from neural progenitor cells [J].
Brito, Catarina ;
Simao, Daniel ;
Costa, Ines ;
Malpique, Rita ;
Pereira, Cristina I. ;
Fernandes, Paulo ;
Serra, Margarida ;
Schwarz, Sigrid C. ;
Schwarz, Johannes ;
Kremer, Eric J. ;
Alves, Paula M. .
METHODS, 2012, 56 (03) :452-460
[5]   An Update on Canine Adenovirus Type 2 and Its Vectors [J].
Bru, Thierry ;
Salinas, Sara ;
Kremer, Eric J. .
VIRUSES-BASEL, 2010, 2 (09) :2134-2153
[6]   Preparation strategy and illumination of three-dimensional cell cultures in light sheet-based fluorescence microscopy [J].
Bruns, Thomas ;
Schickinger, Sarah ;
Wittig, Rainer ;
Schneckenburger, Herbert .
JOURNAL OF BIOMEDICAL OPTICS, 2012, 17 (10)
[7]  
Edelstein Arthur, 2010, Curr Protoc Mol Biol, VChapter 14, DOI 10.1002/0471142727.mb1420s92
[8]   Rapid 3D light-sheet microscopy with a tunable lens [J].
Fahrbach, Florian O. ;
Voigt, Fabian F. ;
Schmid, Benjamin ;
Helmchen, Fritjof ;
Huisken, Jan .
OPTICS EXPRESS, 2013, 21 (18) :21010-21026
[9]   Variational Algorithms to Remove Stationary Noise: Applications to Microscopy Imaging [J].
Fehrenbach, Jerome ;
Weiss, Pierre ;
Lorenzo, Corinne .
IEEE TRANSACTIONS ON IMAGE PROCESSING, 2012, 21 (10) :4420-4430
[10]   Basic building units and properties of a fluorescence single plane illumination microscope [J].
Greger, K. ;
Swoger, J. ;
Stelzer, E. H. K. .
REVIEW OF SCIENTIFIC INSTRUMENTS, 2007, 78 (02)