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 has up to now been characterized by recording an image stack comprising tens of thousands of images and time-consuming post-processing of the data to determine the position of each molecule (reconstruction).
ReconFlex facilitates an on-the-fly data analysis for localization microscopy providing superresolved images in real-time.
It offers ultra-high flexibilty in terms of its two different modes:
Normal Camera Mode: For Live Imaging, adjusting the Microscope and recording Imaging Stacks
Reconstruction Mode: The Localization of the Molecules is being determined by the Camera itself
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 as there is the Possibility of implementing User-Developed Algorithms
Provided with ReconFlex, nanoFLeye opens up a new door to real-time, easy and user-friendly superresolving localization microscopy.
Biomedical as well as material science applications due to flexible sample holder as well as bottom-up setup
Easy sample preparation due to usage of conventional fluorescent dyes
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
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.
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