scholarly journals Theoretical study on Wide-field Fluorescence Microscopy with Dynamic Speckle Illumination

2021 ◽  
Vol 0 (0) ◽  
pp. 0-0
Author(s):  
Keyword(s):  
Fluorescence Microscopy ◽  
Theoretical Study ◽  
Wide Field ◽  
Dynamic Speckle

scholarly journals Recurrent neural network-based volumetric fluorescence microscopy

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Luzhe Huang ◽  
Hanlong Chen ◽  
Yilin Luo ◽  
Yair Rivenson ◽  
Aydogan Ozcan
Keyword(s):  
Neural Network ◽  
Image Reconstruction ◽  
Fluorescence Microscopy ◽  
Recurrent Network ◽  
Sample Volume ◽  
Depth Of Field ◽  
Objective Lens ◽  
Wide Field ◽  
Volumetric Imaging ◽  
3D Image Reconstruction

AbstractVolumetric imaging of samples using fluorescence microscopy plays an important role in various fields including physical, medical and life sciences. Here we report a deep learning-based volumetric image inference framework that uses 2D images that are sparsely captured by a standard wide-field fluorescence microscope at arbitrary axial positions within the sample volume. Through a recurrent convolutional neural network, which we term as Recurrent-MZ, 2D fluorescence information from a few axial planes within the sample is explicitly incorporated to digitally reconstruct the sample volume over an extended depth-of-field. Using experiments on C. elegans and nanobead samples, Recurrent-MZ is demonstrated to significantly increase the depth-of-field of a 63×/1.4NA objective lens, also providing a 30-fold reduction in the number of axial scans required to image the same sample volume. We further illustrated the generalization of this recurrent network for 3D imaging by showing its resilience to varying imaging conditions, including e.g., different sequences of input images, covering various axial permutations and unknown axial positioning errors. We also demonstrated wide-field to confocal cross-modality image transformations using Recurrent-MZ framework and performed 3D image reconstruction of a sample using a few wide-field 2D fluorescence images as input, matching confocal microscopy images of the same sample volume. Recurrent-MZ demonstrates the first application of recurrent neural networks in microscopic image reconstruction and provides a flexible and rapid volumetric imaging framework, overcoming the limitations of current 3D scanning microscopy tools.

scholarly journals Low-Light Photodetectors for Fluorescence Microscopy

2021 ◽  
Vol 11 (6) ◽  
pp. 2773
Author(s):  
Hiroaki Yokota ◽  
Atsuhito Fukasawa ◽  
Minako Hirano ◽  
Toru Ide
Keyword(s):  
Fluorescence Microscopy ◽  
Fluorescence Imaging ◽  
Single Molecule ◽  
Science Studies ◽  
Single Photon ◽  
Laser Scanning Microscopy ◽  
Oxide Semiconductor ◽  
Wide Field ◽  
Single Molecule Fluorescence ◽  
Low Light

Over the years, fluorescence microscopy has evolved and has become a necessary element of life science studies. Microscopy has elucidated biological processes in live cells and organisms, and also enabled tracking of biomolecules in real time. Development of highly sensitive photodetectors and light sources, in addition to the evolution of various illumination methods and fluorophores, has helped microscopy acquire single-molecule fluorescence sensitivity, enabling single-molecule fluorescence imaging and detection. Low-light photodetectors used in microscopy are classified into two categories: point photodetectors and wide-field photodetectors. Although point photodetectors, notably photomultiplier tubes (PMTs), have been commonly used in laser scanning microscopy (LSM) with a confocal illumination setup, wide-field photodetectors, such as electron-multiplying charge-coupled devices (EMCCDs) and scientific complementary metal-oxide-semiconductor (sCMOS) cameras have been used in fluorescence imaging. This review focuses on the former low-light point photodetectors and presents their fluorescence microscopy applications and recent progress. These photodetectors include conventional PMTs, single photon avalanche diodes (SPADs), hybrid photodetectors (HPDs), in addition to newly emerging photodetectors, such as silicon photomultipliers (SiPMs) (also known as multi-pixel photon counters (MPPCs)) and superconducting nanowire single photon detectors (SSPDs). In particular, this review shows distinctive features of HPD and application of HPD to wide-field single-molecule fluorescence detection.

A Single-Molecule Homogeneous Immunoassay by Counting Spatially “Overlapping” Two-Color Quantum Dots with Wide-Field Fluorescence Microscopy

ACS Sensors ◽  
2018 ◽  
Vol 3 (12) ◽  
pp. 2644-2650 ◽  
Author(s):  
Xiaojun Liu ◽  
Conghui Huang ◽  
Chenghua Zong ◽  
Aiye Liang ◽  
Zhangjian Wu ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Wide Field ◽  
Homogeneous Immunoassay

Mapping Pixel Dissimilarity in Wide-Field Super-Resolution Fluorescence Microscopy

2015 ◽  
Vol 87 (9) ◽  
pp. 4675-4682 ◽  
Author(s):  
Cyril Ruckebusch ◽  
Romain Bernex ◽  
Franco Allegrini ◽  
Michel Sliwa ◽  
Johan Hofkens ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Super Resolution ◽  
Wide Field

scholarly journals Wide Field-of-View On-Chip Talbot Fluorescence Microscopy for Longitudinal Cell Culture Monitoring from within the Incubator

2013 ◽  
Vol 85 (4) ◽  
pp. 2356-2360 ◽  
Author(s):  
Chao Han ◽  
Shuo Pang ◽  
Danielle V. Bower ◽  
Patrick Yiu ◽  
Changhuei Yang
Keyword(s):  
Cell Culture ◽  
Fluorescence Microscopy ◽  
Field Of View ◽  
Wide Field ◽  
Wide Field Of View ◽  
On Chip ◽  
Longitudinal Cell ◽  
Culture Monitoring

scholarly journals Mitochondrial transmission during mating in Saccharomyces cerevisiae is determined by mitochondrial fusion and fission and the intramitochondrial segregation of mitochondrial DNA.

1997 ◽  
Vol 8 (7) ◽  
pp. 1233-1242 ◽  
Author(s):  
J Nunnari ◽  
W F Marshall ◽  
A Straight ◽  
A Murray ◽  
J W Sedat ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Fluorescence Microscopy ◽  
Fluorescent Dyes ◽  
Mitochondrial Protein ◽  
Mitochondrial Proteins ◽  
Dimensional Structure ◽  
Wide Field ◽  
Protein Markers ◽  
Protein Marker ◽  
Mitochondrial Transmission

To gain insight into the process of mitochondrial transmission in yeast, we directly labeled mitochondrial proteins and mitochondrial DNA (mtDNA) and observed their fate after the fusion of two cells. To this end, mitochondrial proteins in haploid cells of opposite mating type were labeled with different fluorescent dyes and observed by fluorescence microscopy after mating of the cells. Parental mitochondrial protein markers rapidly redistributed and colocalized throughout zygotes, indicating that during mating, parental mitochondria fuse and their protein contents intermix, consistent with results previously obtained with a single parentally derived protein marker. Analysis of the three-dimensional structure and dynamics of mitochondria in living cells with wide-field fluorescence microscopy indicated that mitochondria form a single dynamic network, whose continuity is maintained by a balanced frequency of fission and fusion events. Thus, the complete mixing of mitochondrial proteins can be explained by the formation of one continuous mitochondrial compartment after mating. In marked contrast to the mixing of parental mitochondrial proteins after fusion, mtDNA (labeled with the thymidine analogue 5-bromodeoxyuridine) remained distinctly localized to one half of the zygotic cell. This observation provides a direct explanation for the genetically observed nonrandom patterns of mtDNA transmission. We propose that anchoring of mtDNA within the organelle is linked to an active segregation mechanism that ensures accurate inheritance of mtDNA along with the organelle.

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