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
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
nanoFLeye is equipped with the pioneering ReconFlex camera developed by Surface Concept GmbH making localization microscopy substantially easier and faster.
Localization microscopy in general is time and disk space consuming by recording an image stack comprising tens of thousands of images for post-processing of the data to determine the position of each molecule (reconstruction).
ReconFlex supports a much faster data analysis for localization microscopy.
It offers ultra-high flexibilty in terms of its different modes:
Normal Camera Mode: For Live Imaging, adjusting the Microscope and recording Imaging Stacks
Reconstruction Support Mode: A Fast Detection Mode of Localization Areas by the nanoFLeye System
Both modes can also be applied simultaneously.
Enormous Reduction in Data Transfer, Time and Disk Space
Select Fiducial Markers prior to the Measurement to correct Possible Drifts in Real-Time
The User retains Full Control over the Algorithms used in the Reconstruction Mode
Provided with ReconFlex, nanoFLeye opens up a new door to real-time, easy and user-friendly superresolving localization microscopy.
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.
Superresolved image of the identical sample position recorded by nanoFleye. The scale bar (horizontal line bottom left) corresponds to 1µm.
Linescan (vertical line in the upper image) of the epifluorescent image.
Linescan (vertical line in the upper image) of the superresolved 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
The images show the pf4 distribution inside the platlets
nanoFLeye uncovers the number of the labeled cytokines as well as their formation in the platelets
Conventional epifluorescent microscopy image of Alexa680-labeled HuPLTs. The image size is 5µm x 5µm.
Superresolved image of the identical sample position recorded by nanoFLeye.
Protein-Polymers in a Tissue Sample
Epifluorescence (30µm x 30µm),
scale bar 5µm
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
SPDM of the identical section,
scale bar 1µm
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 ~200 nm from each other. In a confocal setup it is possible to improve the resolution slightly, but not sufficient to detect single molecules.
Spectral features are used to achieve optical isolation
In Conventional Fluorescence Microscopy the FWHM of the Point-Spread-Function (PSF) >200 nm. 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 20 nm
All localizations found in a stack of usually ten thousands of images are displayed in a single reconstructed super-resolved image