Spectral Characterization of Images
White Light Laser Confocal Microscope Leica TCS SP8 X
Optimization of multi-color image (1)
Primary culture of rat cortical neurons, excitation and detection settings were adjusted to optimal image contrast.
| Structure | Label | Excitation [nm] | Emission[nm] |
| Nucleus | DAPI | 405 | 424-465 |
| Nestin (neuronal stem cells) | cy2 | 495 | 504-545 |
| DCX (immature neurons) | cy3 | 560 | 559-624 |
| Beta III-tubulin | cy5 | 652 | 653-716 |
Optimization of multi-color image (2)
Paramecium stained with 7 dyes excitable in the visible range. Good separation and high image contrast for all 7 colors are possible due to the tunable excitation wavelengths.
Courtesy of Paolo Bianchini & Alberto Diaspro, Istituto Italiano di Tecnologia, Genoa, Italy
Removal of back scattered light by LightGate
LightGate completely removes back scattered light.
Sample: fixed HeLa cells, tubulin stained with BD Horizon V-500, nucleus stained with Chromeo 505. Images are recorded directly under excitation wavelength.
Full spectral characterization using LightGate
The rectangular lambda square plot displays full spectral information. This information is useful to investigate samples with complex or unknown fluorescence and to characterize new developed fluorophores.
The grey line within the graphical definition window indicates excitation = detection.