Confocal laserscanning microscopy laboratory
Documenting, quantifying and describing morphological characters of species represent the indispensable basis for any taxonomic, systematic and morphological research. A wide variety of imaging techniques for the study of marine protists, plants and animals are available today.
Confocal laserscanning microscopy (CLSM) is an ideal tool for the study of microscopic organisms from the marine benthic and pelagic realms. In the CLSM-laboratory of the German Centre for Marine Biodiversity Research (DZMB) we frequently use the system for imaging the external morphology of micro-crustaceans and reconstructing internal organ systems of soft-bodied meiofaunal taxa.
For this purpose, different histochemical and immune-histochemical staining procedures are applied. We also aim to develop and improve further areas of application and preparation techniques such as the amplification of cuticular autofluorescence, or the imaging and examination of micro-bioeroding organisms.The CLSM laboratory of our department is equipped with a Leica® TCS SP5 system coupled to a DM5000B microscope basic unit. Six visible laser wavelengths (458, 476, 488, 514, 561 and 633 nm), two fluorescence detectors and one further detector for the transmitted light channel are available for our experiments. The fluorescent objects get excited with a diffraction-limited focused laser beam that is scanned line-by-line over the specimen. The confocal collocation of two variable apertures allows the elimination of so-called ‘out-of-focus’ signals. A stepwise shift of the microscopic stage finally enables the compilation of three-dimensional image stacks. Such datasets can be processed manifold ranging from generating projection images with an extended depth of focus to three-dimensional reconstructions of the external morphology and internal anatomy of the examined organisms, including quantifications such as the measuring of the biovolume of organ systems.