nanoFLeye™

The Way to a Deeper Insight into the Cell

nanoFLeye™ (nanoFluorescenceEye) is the innovative reply on demands and needs in the field of superresolving optical imaging based on the localization microscopy technique SPDM (spectral precision distance microscopy).

nanoFLeye™ excels with

3D Multicolour Imaging based on SPDM

High Stability

High Flexibility

Choose up to Four Different Excitation Wavelengths Suitable to your Desired Dyes

Choose your Favoured Microscope Objective

Clear and User-Friendly Interface

Possibility of Remote Control

Opportunity of High Level Automatization such as Autofocus, Easily Programmable Measurement Sequences and Automated Data Analysis

APPLICATION EXAMPLES

nanoFLeye™ allows to reveal structures well below the Abbe limit being of interest for biomedical as well as material science applications.

The inverted setup of nanoFLeye™ and its flexible sample holder allows for the investigation of a huge variety of sample configurations.

 

Protein-Polymers in a Cell Culture Specimen

The images show a segment of fluorescence-labeled microtubules in a cell culture (HeLa-cells)

nanoFLeye™ uncovers sub-structures within the polymer configuration

The corresponding linescans show the intensity distribution along the implied line in the images

Conventional epifluorescent microscopy image of Alexa647-labeled microtubuli of HeLa-cells; scale bar 1µm.

Super-resolved image of the identical sample position recorded by nanoFleye™; scale bar 1µm.

Linescan (vertical line in the upper image) of the epifluorescent image.

Linescan (vertical line in the upper image) of the super-resolved image.

Alexa680-labeled human platelets (HuPLTs, PF4, A680, native)

sample preparation courtesy of: Dr. M. Schmitt, LMU München

left: conventional epifluorescent microscopy image

right: superresolved image recorded by nanoFLeye™

Human Thrombocytes

The images show the PF4 distribution inside the platelets

nanoFLeye™ uncovers the number of the labeled cytokines as well as their formation in the platelets

Conventional epifluorescent microscopy image of Alexa680-labeled HuPLTs; scale bar 1µm.

Super-resolved image of the identical sample position recorded by nanoFLeye™; scale bar 1µm.

Protein-Polymers in a Tissue Sample

Epifluorescence (30µm x 30µm),

scale bar 5µm

Immunofluorescence micrographs

of a 30µm thick rat brain tissue

ICV 10 Hippocampus (DG)

Staining: microtubules (Alexa647)

The images show a segment of fluorescence-labeled microtubules in 30µm thick rat brain tissue

The histogram shows the distribution of the localization accuracy of the detected events.

A mean localzation accuracy of 18nm could be achieved

Mean localization accuracy: 18nm

Sample preparation courtesy of:

Dr. Sebastian Bauer

Leiter AG Translationale Epileptologie

Epilepsiezentrum Frankfurt Rhein-Main

Klinik für Neurologie

Goethe Universität Frankfurt

Epifluorecence; scale bar 1µm.

Super-resolved image of the identical section; scale bar 1µm.

METHOD

Convential fluorescence microscopy is a versatile tool to perform functional cell biology analysis. Fluorophores are being coupled to antibodies which bind to their corresponding proteins in the cell. By analyzing the fluorescence signals in the microscopy image, one can get insight to  the distribution of the chosen proteins inside the cell.

 

However, in terms of nano-science and detailed insight into biological processes on a molecular level, convential fluorescence microscopy is stretched to its limit.

 

In an epifluorescence microscope the lateral resolution is determined by the diffraction limit, i.e. you cannot distinguish two molecules having a distance less than ~200nm from each other. In a confocal setup it is possible to improve the resolution slightly, but not sufficient to detect single molecules.

Localization Microscopy

Spectral features are used to achieve optical isolation

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In Conventional Fluorescence Microscopy the Full-Width-at-Half-Maximum (FWHM) of the Point-Spread-Function (PSF) >200nm. Signals of adjacent dyes overlap, therefore single molecules cannot be resolved

 

Using SPDM, randomly activated dyes are „optically isolated“, i.e. no overlap of the signal of adjacent molecules can occur

 

The locations of the optically isolated fluorophores are determined by the localization algorithm with a precision down to 20nm

 

All localizations found in a stack of usually ten thousands of images are displayed in a single reconstructed super-resolved image

SPECIAL FEATURE

nanoFLeye™ is equipped with the pioneering ReconFlex™ camera developed by Surface Concept GmbH making localization microscopy substantially easier and faster.

 

For more information, see here.

 

CONTACT

Surface Concept GmbH

 

Am Sägewerk 23a

55124 Mainz

Germany

 

phone: +49 6131 62716 0

fax:        +49 6131 62716 29

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3D Multicolour Imaging based on SPDM

High Stability

High Flexibility

Clear and User-Friendly Interface

Possibility of Remote Control

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