Fluorescence

Fluorescence itself is a form of luminescence that results from matter emitting light of a certain wavelength after absorbing electromagnetic radiation. Fluorescence Imaging help visualize biological processes taking place in a living organism

Intelligent Acquisition System

With Nikon microscope and software NIS-Elements, our high content imaging streamlines high-speed imaging and simple operations.

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Content

Related Products

Microscopy

Nikon Super Resolution N-SIM
Nikon Super Resolution N-STORM
Nikon Confocal Microscope System
AX / AX R
Nikon Spinning Disk Confocal
Nikon Inverted Microscope
Ti2-E
MIGHTEX Calcium Imaging and Optogenetics
OASIS Polygon
Tomocube Holotomographic Microscopy
HT Series

Cameras

PCO. Fluoresence Lifetime Imaging Cameras
FLIM
Andor Scientific Fluorescence Camera
SONA
Nikon Scientific Monochrome Cameras
DS-Qi2
TELEDYNE PHOTOMETRICS Scientific Monochrome Cameras
KINETIX

Software

Nikon NIS-Elements HC
Nikon NIS-Elements AR

Application Examples

Cell and Molecular Biology  

Automatic morphometric extraction of the rat osteocytic lacunae using Segment.ai

Osteocytes form an intercellular network similar to a neuronal network in bone. The research team visualizes and measures this network structure, the functional significance of which remains to be elucidated. Using Nikon NIS-Elements' Segment.ai, osteocytic lacunae are automatically segmented in order to facilitate the measurement of their numbers and morphology.

Network of osteocytic lacunae-canalicular system

Auto-fluorescence imaging of Villanueva-stained rabbit tibia. This image was taken as a Z stack with a resolution of 0.04 um/pixel using a 100X objective, subjected to enhanced resolution processing, and displayed as a maximum intensity projection (MIP). Scale bar: 20 um

Villanueva-stained un-decalcified bone section of a rabbit 

Results of segmentation by the conventional binarization method (blue). Structures other than osteocytic lacunae, such as osteocytic canaliculi and some parts of bone marrow, were detected (arrows). Manual mask modification or removal is required for accurate measurement.

Villanueva-stained un-decalcified bone section of a rabbit 

Results of segmentation using Segment.ai (orange). Only the osteocytic lacunae were fully detected by Segment.ai, which learned how to identify osteocytic lacuna areas.

AI Tool Involved

Some images are nearly impossible to segment by traditional intensity thresholding methods. A neural network can be trained by human classification of structures of interest that cannot easily be defined by classic thresholding and image processing by using Segment.ai.

By tracing features of interest and training these compared to the underlying image, the neural network can learn and apply segmentation to similar images, recognizing features previously only identifiable by painstaking manual tracing approaches. Learn More>>

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Microscope Involved

Since a resonant scanner can perform confocal imaging with higher temporal resolution than a Galvano scanner, it is used in many cases to acquire life phenomena occurring at high speeds. In contrast, because the resonant scanner of the new generation AX R confocal microscope system supports up to 2K x 2K acquisition, it can be used for a wide range of purposes, from high-speed imaging to high-resolution imaging. Learn More>>

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Renal Biopsy

Quantitative analysis of lesions of renal foot processes and mitochondria by Structured Illumination Super-Resolution Microscope

In this study, Dr. Matsumoto et al. established a method for quantifying observed microstructural changes using the Fourier transform. When human renal biopsy samples were evaluated using this method, the degree of disruption of glomerular epithelial cell foot processes was correlated with the amount of urinary protein 

Obstacle

Although an electron microscope is usually required to visualize these microstructural changes, most renal biopsy tissue is prepared as an optical microscope specimen, and the specimens observed with an electron microscope are so small that important lesions could be overlooked. Establishing a method for evaluating these lesions over the entire collected tissue using an optical microscope specimen has therefore been anticipated.

Visualization and quantification of human glomerular epithelial cell structure disruption in EMT-stained sections

In patients with lgA nephropathy, it was observed that the higher the amount of urinary protein, the higher the degree of structural disruption of the foot process.

Visualization of glomerular epithelial cell structure disruption and mitochondrial damage in animal models

In a mouse model of tubular stromal disorders due to LPS administration or ischemia-reperfusion injury, mitochondrial fragmentation and swelling in tubular epithelial cells werw observed.

Microscope Involved

The N-SIM S utilizes Structured Illumination Microscopy (SIM) technology to capture the minute structures within a specimen at twice the resolution of conventional light microscopes.

- Lateral resolution: 115 nm (3D-SIM mode),
86 nm (TIRF-SIM mode)
- Axial resolution: 269 nm (3D-SIM mode)
- Field of view: Up to 66 um x 66 um (with a
100X objective)

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*Photos courtesy of Nikon official website

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