Primer: fluorescence imaging under the diffraction limit

2008 ◽  
Vol 6 (1) ◽  
pp. 19-20 ◽  
Author(s):  
Daniel Evanko
Keyword(s):  
Fluorescence Imaging ◽  
Diffraction Limit

scholarly journals Efficiency-enhanced and sidelobe-suppressed super-oscillatory lenses for sub-diffraction-limit fluorescence imaging with ultralong working distance

Nanoscale ◽  
2020 ◽  
Vol 12 (13) ◽  
pp. 7063-7071 ◽  
Author(s):  
Wenli Li ◽  
Pei He ◽  
Weizheng Yuan ◽  
Yiting Yu
Keyword(s):  
Fluorescence Imaging ◽  
Diffraction Limit ◽  
Working Distance

Customized efficiency-enhanced and sidelobe-suppressed super-oscillatory lenses for sub-diffraction-limit fluorescence imaging with ultralong working distances without photobleaching and a pinhole filter.

scholarly journals SOFIevaluator: a strategy for the quantitative quality assessment of SOFI data

2019 ◽  
Author(s):  
Benjamien Moeyaert ◽  
Wim Vandenberg ◽  
Peter Dedecker
Keyword(s):  
Fluorescence Imaging ◽  
Computational Analysis ◽  
Fluorescent Proteins ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Imaging Data ◽  
Sample Structure ◽  
Imaging Conditions

AbstractSuper-resolution fluorescence imaging techniques allow optical imaging of specimens beyond the diffraction limit of light. Super-resolution optical fluctuation imaging (SOFI) relies on computational analysis of stochastic blinking events to obtain a super-resolved image. As with some other super-resolution methods, this strong dependency on computational analysis can make it difficult to gauge how well the resulting images reflect the underlying sample structure. We herein report SOFIevaluator, an unbiased and parameter-free algorithm for calculating a set of metrics that describes the quality of super-resolution fluorescence imaging data for SOFI. We additionally demonstrate how SOFIevaluator can be used to identify fluorescent proteins that perform well for SOFI imaging under different imaging conditions.

scholarly journals Superaufgelöste Mikroskopie: Pilze unter Beobachtung

BIOspektrum ◽  
2021 ◽  
Vol 27 (4) ◽  
pp. 380-382
Author(s):  
Sebastian Sputh ◽  
Sabine Panzer ◽  
Christian Stigloher ◽  
Ulrich Terpitz
Keyword(s):  
Electron Microscopy ◽  
Membrane Protein ◽  
Fluorescence Imaging ◽  
Filamentous Fungus ◽  
Light And Electron Microscopy ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Fusarium Fujikuroi ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

AbstractThe diffraction limit of light confines fluorescence imaging of subcellular structures in fungi. Different super-resolution methods are available for the analysis of fungi that we briefly discuss. We exploit the filamentous fungus Fusarium fujikuroi expressing a YFP-labeled membrane protein showing the benefit of correlative light- and electron microscopy (CLEM), that combines structured illumination microscopy (SIM) and scanning election microscopy (SEM).

scholarly journals Adapting the 3D-printed Openflexure microscope enables computational super-resolution imaging

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 2003
Author(s):  
Stephen D. Grant ◽  
Gemma S. Cairns ◽  
Jordan Wistuba ◽  
Brian R. Patton
Keyword(s):  
Fluorescence Imaging ◽  
Low Cost ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Entire System ◽  
3D Printed ◽  
Resolution Imaging ◽  
Cost Components ◽  
Better Than

We report on a 3D printed microscope, based on a design by the Openflexure project, that uses low cost components to perform fluorescence imaging. The system is sufficiently sensitive and mechanically stable to allow the use of the Super Resolution Radial Fluctuations algorithm to obtain images with resolution better than the diffraction limit. Due to the low-cost components, the entire system can be built for approximately $1200.

Confocal Raman mapping

1992 ◽  
Vol 50 (2) ◽  
pp. 1514-1515
Author(s):  
J. Barbillat ◽  
M. Delhaye ◽  
P. Dhamelincourt
Keyword(s):  
Chemical Species ◽  
Diffraction Limit ◽  
Microscopic Level ◽  
Raman Mapping ◽  
Complete Spectrum ◽  
Laser Illumination ◽  
Ccd Detector ◽  
Raman Microprobe ◽  
Photodiode Array ◽  
Raman Image

Raman mapping, with a spatial resolution close to the diffraction limit, can help to reveal the distribution of chemical species at the surface of an heterogeneous sample.As early as 1975,three methods of sample laser illumination and detector configuration have been proposed to perform Raman mapping at the microscopic level (Fig. 1),:- Point illumination:The basic design of the instrument is a classical Raman microprobe equipped with a PM tube or either a linear photodiode array or a two-dimensional CCD detector. A laser beam is focused on a very small area ,close to the diffraction limit.In order to explore the whole surface of the sample,the specimen is moved sequentially beneath the microscope by means of a motorized XY stage. For each point analyzed, a complete spectrum is obtained from which spectral information of interest is extracted for Raman image reconstruction.- Line illuminationA narrow laser line is focused onto the sample either by a cylindrical lens or by a scanning device and is optically conjugated with the entrance slit of the stigmatic spectrograph.

The influence of confocal microscope design on imaging performance

1993 ◽  
Vol 51 ◽  
pp. 144-145
Author(s):  
C J R Sheppard
Keyword(s):  
Image Quality ◽  
Fluorescence Imaging ◽  
Confocal Microscope ◽  
Industrial Applications ◽  
Three Dimensions ◽  
Design Parameters ◽  
Weak Signals ◽  
Size And Shape ◽  
Imaging Performance ◽  
Fundamental Limitation

The confocal microscope is now widely used in both biomedical and industrial applications for imaging, in three dimensions, objects with appreciable depth. There are now a range of different microscopes on the market, which have adopted a variety of different designs. The aim of this paper is to explore the effects on imaging performance of design parameters including the method of scanning, the type of detector, and the size and shape of the confocal aperture.It is becoming apparent that there is no such thing as an ideal confocal microscope: all systems have limitations and the best compromise depends on what the microscope is used for and how it is used. The most important compromise at present is between image quality and speed of scanning, which is particularly apparent when imaging with very weak signals. If great speed is not of importance, then the fundamental limitation for fluorescence imaging is the detection of sufficient numbers of photons before the fluorochrome bleaches.

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