Three-Dimensional Live Imaging of Filamentous Fungi with Light Sheet-Based Fluorescence Microscopy (LSFM)

Author(s):  
Francesco Pampaloni ◽  
Laura Knuppertz ◽  
Andrea Hamann ◽  
Heinz D. Osiewacz ◽  
Ernst H. K. Stelzer
Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1255
Author(s):  
Norio Yamashita ◽  
Masahiko Morita ◽  
Hideo Yokota ◽  
Yuko Mimori-Kiyosue

From cells to organisms, every living system is three-dimensional (3D), but the performance of fluorescence microscopy has been largely limited when attempting to obtain an overview of systems’ dynamic processes in three dimensions. Recently, advanced light-sheet illumination technologies, allowing drastic improvement in spatial discrimination, volumetric imaging times, and phototoxicity/photobleaching, have been making live imaging to collect precise and reliable 3D information increasingly feasible. In particular, lattice light-sheet microscopy (LLSM), using an ultrathin light-sheet, enables whole-cell 3D live imaging of cellular processes, including mitosis, at unprecedented spatiotemporal resolution for extended periods of time. This technology produces immense and complex data, including a significant amount of information, raising new challenges for big image data analysis and new possibilities for data utilization. Once the data are digitally archived in a computer, the data can be reused for various purposes by anyone at any time. Such an information science approach has the potential to revolutionize the use of bioimage data, and provides an alternative method for cell biology research in a data-driven manner. In this article, we introduce examples of analyzing digital mitotic spindles and discuss future perspectives in cell biology.


2014 ◽  
Vol 6 (10) ◽  
pp. 988-998 ◽  
Author(s):  
Francesco Pampaloni ◽  
Ulrich Berge ◽  
Anastasios Marmaras ◽  
Peter Horvath ◽  
Ruth Kroschewski ◽  
...  

This novel system for the long-term fluorescence imaging of live three-dimensional cultures provides minimal photodamage, control of temperature, CO2, pH, and media flow.


2020 ◽  
Vol 8 ◽  
Author(s):  
Xiaoyan Li ◽  
Yubing Han ◽  
Wenjie Liu ◽  
Cuifang Kuang ◽  
Xu Liu ◽  
...  

Three-dimensional (3D) imaging using dual-lens fluorescence microscopies is popular in observing fluorescently labeled biological samples, such as mammalian/model animal cells, tissues, and embryos. Specifically, dual-lens super-resolution fluorescence microscopy methods using two opposing objective lenses allow significantly higher axial resolution and better signal to noise ratio than traditional single-lens counterparts, and thus distinguish more details in 3D images of fine intracellular structures. For 3D imaging of thick tissues and entire embryos, dual-lens light-sheet fluorescence microscopy methods using two objective lenses, either orthogonal or non-orthogonal, to achieve selective plane illumination, can meet the requirements, and thus can be used to observe embryo development and structures of interest in thick tissues. This review summarizes both dual-lens fluorescence microscopy methods, including their principles, configurations, and 3D imaging applications, providing a guideline for biological laboratories with different 3D imaging needs.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hosein Kafian ◽  
Meelad Lalenejad ◽  
Sahar Moradi-Mehr ◽  
Shiva Akbari Birgani ◽  
Daryoush Abdollahpour

Abstract Light-sheet fluorescence microscopy (LSFM) has now become a unique tool in different fields ranging from three-dimensional (3D) tissue imaging to real-time functional imaging of neuronal activities. Nevertheless, obtaining high-quality artifact-free images from large, dense and inhomogeneous samples is the main challenge of the method that still needs to be adequately addressed. Here, we demonstrate significant enhancement of LSFM image qualities by using scanning non-diffracting illuminating beams, both through experimental and numerical investigations. The effect of static and scanning illumination with several beams are analyzed and compared, and it is shown that scanning 2D Airy light-sheet is minimally affected by the inhomogeneities in the samples, and provides higher contrasts and uniform resolution over a wide field-of-view, due to its reduced spatial coherence, self-healing feature and longer penetration depth. Further, the capabilities of the illumination scheme is utilized for both single-and double-wavelength 3D imaging of large and dense mammospheres of cancer tumor cells as complex inhomogeneous biological samples.


Lab on a Chip ◽  
2021 ◽  
Author(s):  
Erick Vargas Ordaz ◽  
Sergey Gorelick ◽  
Harrison York ◽  
Bonan Liu ◽  
Michelle L. Halls ◽  
...  

Volumetric, sub-micron to micron level resolution imaging is necessary to assay phenotypes or characteristics at the sub-cellular/organelle scale. However, three-dimensional fluorescence imaging of cells is typically low throughput or compromises...


2015 ◽  
Vol 107 (26) ◽  
pp. 263701 ◽  
Author(s):  
C. K. Rasmi ◽  
Kavya Mohan ◽  
M. Madhangi ◽  
K. Rajan ◽  
U. Nongthomba ◽  
...  

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