Quantification, identification and analysis of fluorescent signals within single cells are a gold standard in biological cell research and numerous clinical applications. Flow cytometry can be used to detect the presence or even the distribution of these molecules at very high speed. Our method of real-time deformability cytometry (RT-DC), for label-free analysis of cell functions reaches similar measurement speeds. Thus, we combined the two methods in RT-FDC, allowing to discover biological and biophysical phenomena in an unprecedented manner.
The module is attached to the side port of the microscope and can be equipped with 3 excitation lasers and 3 detection channels. A confined excitation (light sheet across the channel) allows for 1D fluorescence imaging. In addition to all RT-DC parameters, the fluorescent signals are analyzed (intensity, width, area) in real-time, at a rate of more than 100 cells per second.
Identify blood cells based on established surface markers:
The FluorescenceModule enables the detection and identification of up to three different fluorophore species in one sample. Cell identification and simultaneous evaluation of mechanical and morphological properties becomes feasable with the help of fluorescently labeled surface proteins. The plots above show cell populations within a G-CSF mobilized peripheral blood sample. The detection of the fluorescence intensity of the cell surface markers CD3-FITC (indicating T-cells), CD34-PE (indicating hematopoietic stem cells), and CD14-APC (indicating monocytes) reveals distinct mechanical phenotypes of each cell type.
One-dimensional fluorescence imaging:
The excitation light path in the FluorescenceModule is designed to produce a confined light sheet across the channel, so that cells travel through a narrow curtain of excitation light. This allows for 1D fluorescence imaging, i.e., the lateral distribution of fluorescently labeled structures can be resolved in flow direction. The shape of the detected fluorescence peak carries important information. Fluorescently labeled intracellular structures like the nucleus will appear as narrow peak as compared to peaks resulting from fluorescently labeled cytoplasm. Labeled histones inside a dividing cell, for instance, yield a distinct peak pattern depending on cell cycle stage as shown in the plots above.
The package includes:
The FluorescenceModule in the highest technical configuration (3 Lasers, 3 Detection channels) is 130 000 €.
(only for business customers from academia, excl. VAT)