The confocal laserscanning microscope Leica (R) TCS SP5 we use is ideally suited for the examination of very small marine organisms

Confocal laserscanning microscopy

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 division 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.

Articles of Confocal laserscanning microscopy

50. MATHISKE, A., THISTLE, D., GHEERARDYN, H. & VEIT-KÖHLER, G. (accepted): Deep sea without limits – four new closely related species of Emertonia Wilson, 1932 (Copepoda, Harpacticoida, Paramesochridae) show characters with a world-wide distribution. Zootaxa xxxx: yyy-zzz.

49. MALYUTINA, M.V., KIHARA, T.C. & BRIX, S. (accepted): A new genus of Munnopsidae Lilljeborg, 1864 (Crustacea, Isopoda), with description of two new species from the Clarion Clipperton Fracture zone, north-eastern tropical Pacific. Marine Biodiversity xxx: yyy-zzz.

48. KHODAMI, S., MERCADO-SALAS, N.F. & MARTINEZ ARBIZU, P. (accepted): Genus level molecular phylogeny of Aegisthidae Giesbrecht, 1893 (Copepoda: Harpacticoida) reveals morphological adaptation to deep sea and pelagic habitats. BMC Evolutionary Biology xx: yy.

47. SCHNIER, J., AHLRICHS, W.H., GRUHL, A., SCHULBERT, C., TEICHERT. S. & KIENEKE, A. (2019): Ultrastructure of the epidermal gland system of Tetranchyroderma suecicum Boaden, 1960 (Gastrotricha: Macrodasyida) indicates a defensive function of its exudate. Zoomorphology 138: 443-462.

46. GEORGE, K.H. et al. (2020): Copepoda. In: Schmidt-Rhaesa, A. (ed.): Guide to the Identification of Marine Meiofauna, pp. 465-533, Verlag Dr. Friedrich Pfeil, München.

45. KIENEKE, A., MÜNTER, L. & RIEMANN, O. (2020): Gastrotricha. In: Schmidt-Rhaesa, A. (ed.): Guide to the Identification of Marine Meiofauna, pp. 104-163, Verlag Dr. Friedrich Pfeil, München.

44. ROSSEL, S. & MARTÍNEZ ARBIZU, P. (2019): Revealing higher than expected diversity of Harpacticoida (Crustacea: Copepoda) in the North Sea using MALDI-TOF MS and molecular barcoding. Nature Scientific Reports 9: 9182 DOI:

43. GOMEZ, S. (accepted): 21. Class Copepoda. Harpacticoida. In: Damborenea, C., Rogers, D.C., Thorp, J.H. (eds.): Thorp and Covich’s Freshwater Invertebrates. Fourth Edition. Volume V. Keys to Neotropical and Antarctic Fauna, pp. xxx-yyy, Academic Press, New York.

42. SEPAHVAND, V., KIHARA, T.C. & BOXSHALL, G. (2019): A new species of Clausidium Kossmann, 1874 (Copepoda: Cyclopoida) associated with ghost shrimps from the Persian Gulf, including female-male interlocking mechanisms and remarks on host specificity. Sytematic Parasitology 96: 171-189 DOI: 10.1007/s11230-019-09839-x

41. MERCADO-SALAS, N. F., KHODAMI, S. KIHARA, T.C., ELÍAS-GUTIÉRREZ, M. & MARTÍNEZ ARBIZU, P. (2018): Genetic structure and distributional patterns of the genus Mastigodiaptomus (Copepoda) in Mexico, with the description of a new species from the Yucatan Peninsula. Arthropod Systematics & Phylogeny 76: 487-507.

40. MERCADO-SALAS, N.F., KHODAMI, S. & MARTÍNEZ ARBIZU, P. (2019): Convergent evolution of mouthparts morphology between Siphonostomatoida and a new genus of deep-sea Aegisthidae Giesbrecht, 1893 (Copepoda: Harpacticoida). Marine Biodiversity 49: 1635-1655. DOI: 10.1007/s12526-018-0932-3

39. CORGOSINHO, P.H., KIHARA, T.C., NIKOLAOS V. SCHIZAS, N.V., ALEXANDRA OSTMANN, A., MARTÍNEZ ARBIZU, P. & IVANENKO, V. (2018): Traditional and confocal description of a new genus and two new species of deep water Cerviniinae Sars, 1903 (Copepoda, Harpacticoida, Aegisthidae) from the Southern Atlantic and the Norwegian Sea. Zookeys 766: 1-38.

38. BRIX, S., BOBER, S., TSCHESCHE, C., KIHARA, T.C., DRISKELL, A. & JENNINGS, R.M. (2018): Molecular species delimitation and its implications for species descriptions using desmosomatid and nannoniscid isopods from the VEMA fracture zone as example taxa. Deep Sea Research Part II: Topical Studies in Oceanography 148: 180-207. DOI: 10.1016/j.dsr2.2018.02.004

37. BRANDT, A., SCHOLZ, J., ALLSPACH, A., BRENKE, N., BRIX, S., GEORGE, K.H., HÖRNSCHEMEYER, T., HOLST, S., HOPPENRATH, M., IWAN, F., JANSSEN, A., JANSSEN, R., JANUSSEN, D., JESKULKE, K., FIEGE, D., KAISER, S., KIENEKE, A., KIHARA, T.C., KRÖNCKE, I., KRUPP, F., MARTHA, S.O., MARTÍNEZ ARBIZU, P.M., MEIßNER, K., MILJUTINA, M., MILJUTIN, D., RENZ, J., RIEHL, T., SAEEDI, H., SIEGLER, V., SONNEWALD, M., STUCKAS, H. & VEIT-KÖHLER, G. (2018): 200 years of marine research at Senckenberg: selected highlights. Marine Biodiversity 48: 159-178.

36. SEPAHVAND, V., RASTEGAR-POUYANI, N. & KIHARA, T.C. (2018): Two new species of Clausidium copepods (Crustacea, Poecilostomatoida) associated with ghost shrimps from Iran. Journal of the Marine Biological Association of the United Kingdom 98(6): 1401-1409. DOI: 10.1017/S0025315417000303

35. SEPAHVAND, V., RASTEGAR-POUYANI, N., KIHARA, T.C. & MOMTAZI, F. (2017): A new species of Clausidium Kossmann, 1874 (Crustacea, Copepoda,Cyclopoida, Clausidiidae) associated with ghost shrimps from Iran. Nauplius 25: e2017018. DOI: 10.1590/2358-2936e2017018

34. MEIßNER, K., BICK, A. & GÖTTING, M. (2017): Arctic Pholoe (Polychaeta, Pholoidae): when integrative taxonomy helps to sort out barcodes. Zoological Journal of the Linnean Society 179: 237-262. Doi: 10.1111/zoj.12468

33. KAISER, S., BRIX, S., KIHARA, T.C., JANSSEN, A. & JENNINGS, R.M. (2018): Integrative species delimitation in the deep-sea genus Thaumastosoma Hessler, 1970 (Isopoda, Asellota, Nannoniscidae) reveals a new genus and species from the Atlantic and central Pacific abyss. Deep-Sea Research Part II: Topical Studies in Oceanography 148: 151-179.

32. FRICKE, A., KIHARA, T.C. & HOPPENRATH, M. (2017): Studying mesoalgal structures: a non-destructive approach based on confocal laser scanning microscopy. Botanica Marina 60(2): 181-195. DOI 10.1515/bot-2016-0057

31. MÜNTER, L. & KIENEKE, A. (2017): Novel myo-anatomical insights to the Xenotrichula intermedia species complex (Gastrotricha: Paucitubulatina): Implications for a pan-European species and reconsideration of muscle homology among Paucitubulatina. Proceedings of the Biological Society of Washington 130: 165-185.

30. KIENEKE, A. & NIKOUKAR, H. (2017): Integrative morphological and molecular investigation of Turbanella hyalina Schultze, 1853.(Gastrotricha: Macrodasyida), including a redescription of the species. Zoologischer Anzeiger 267: 168-186.

29. KAMANLI, S.A., KIHARA, T.C., BALL, A.D., MORRIT, D. & CLARK, P.F. (2017): A 3D imaging and visualization workflow, using confocal microscopy and advanced image processing for brachyuran crab larvae. Journal of Microscopy 266(3): 307–323. DOI: 10.1111/jmi.12540

28. FRICKE, A., KIHARA, T.C., KOPPRIO, G.A. & HOPPENRATH, M. (2017): Anthropogenically driven habitat formation by a tube dwelling diatom on the Northern Patagonian Atlantic coast. Ecological Indicators 77: 8–13.

27. CORGOSINHO, P.H.C., MERCADO-SALAS, N.F., MARTÍNEZ ARBIZU, P., SILVA, E.N.S. & KIHARA, T.C. (2017): Revision of the Remaneicaris argentina-group (Copepoda, Harpacticoida, Parastenocarididae): supplementary description of species, and description of the first semi-terrestrial Remaneicaris from the tropical forest of Southeast Mexico. Zootaxa 4238 (4): 499–530.

26. BRUCE, N.L., BRIX, S., BALFOUR, N., KIHARA, T.C., WEIGAND, A.M., MEHTERIAN, S. & ILIFFE, T.M. (2017): A new genus for Cirolana troglexuma Botosaneanu & Iliffe, 1997, an anchialine cave dwelling cirolanid isopod (Crustacea, Isopoda, Cirolanidae) from the Bahamas. Subterranean Biology 21: 57–92.

25. VAKATI, V., KIHARA, T.C. & LEE, W. (2016): A new species of the genus Nannopus (Copepoda, Harpacticoida, Nannopodidae) from the mudflat of Ganghwa Island, Korea. Proceedings of the Biological Society of Washington. 129:212–233. DOI: 10.2988/0006-324X-129.Q3.212

24. GOLLNER, S., STUCKAS, H., KIHARA, T.C., LAURENT, S., KODAMI, S. & MARTINEZ ARBIZU, P. (2016): Mitochondrial DNA Analyses Indicate High Diversity, Expansive Population Growth and High Genetic Connectivity of Vent Copepods (Dirivultidae) across Different Oceans. PLoS ONE 11(10): e0163776. doi:10.1371/journal pone.0163776

23. KIENEKE, A. & SCHMIDT-RHAESA, A. (2015): Gastrotricha. In: Schmidt-Rhaesa, A. (ed.) Handbook of Zoology. Gastrotricha, Cycloneuralia and Gnathifera. Volume 3: Gastrotricha and Gnathifera, pp. 1-134. De Gruyter, Berlin.

22. GUGGOLZ, T., HENNE, S., POLITI, Y., SCHÜTZ, R., MASIC, A., MÜLLER, C.H.G. & MEIßNER, K. (2015): Histochemical evidence of β-chitin in parapodial glandular organs and tubes of Spiophanes (Annelida, Sedenteria: Spionidae), and first studies on selected Annelida. Journal of Morphology 276, 1433–1447.

21. BRIX, S., LEESE, F., RIEHL, T. & KIHARA, T.C. (2014): A new genus and new species of Desmosomatidae Sars, 1897 (Isopoda) from the east South-Atlantic abyss described by means of integrative taxonomy. Marine Biodiversity. Marine Biodiversity. 45 (1), 7–61. DOI 10.1007/s12526-014-0218-3

20. BRANDT, A., BRIX, S., HELD, C. & KIHARA, T.C. (2014): Molecular differentiation in sympatry despite morphological stasis: deep-sea Atlantoserolis Wägele, 1994 and Glabroserolis Menzies, 1962 from the south-west Atlantic (Crustacea: Isopoda: Serolidae). Zoological Journal of the Linnean Society. 172, 318–359. DOI: 10.1111/zoj.12178

19. POINTNER, K., KIHARA, T.C., GLATZEL, T. & VEIT-KÖHLER, G. (2013): Two new closely related deep-sea species of Paramesochridae (Copepoda, Harpacticoida) with extremely differing geographical range sizes. Marine Biodiversity 43: 293–319. doi: 10.1007/s12526-013-0158-3

18. KOTTMANN, J., KIHARA, T.C., GLATZEL, T. & VEIT-KÖHLER, G. (2013): A new species of Wellsopsyllus (Copepoda, Harpacticoida, Paramesochridae) from the deep Southern Ocean and remarks on its biogeography. Helgoland Marine Research 67: 33–48. doi: 10.1007/s10152-012-0302-7

17. KIHARA, T.C. & ROCHA, C.E.F. (2013): First record of Clausidium (Copepoda, Clausidiidae) from Brazil: a new species associated with ghost shrimps Neocallichirus grandimana (Gibbes, 1850) (Decapoda, Callianassidae). ZooKeys 335: 47–67. DOI: 10.3897/zookeys.335.5490

16. BJÖRNBERG, T. & KIHARA, T.C. (2013): On Tetragonicipitidae (Crustacea, Copepoda) from the Channel of São Sebastião, Brazil, with description of their nauplii and two new species of Phyllopodopsyllus. Zootaxa 3718 (6): 501–529. DOI:10.11646/zootaxa.3718.6.1

15. WILTS, E.F., D. WULFKEN, W.H. AHLRICHS, & P. MARTÍNEZ ARBIZU. (2012): The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology. Acta Zoologica 93(1): 14–27.

14. MILJUTINA M.A. & MILJUTIN D.M. (2012): Seven new and four known species of the genus Acantholaimus (Nematoda: Chromadoridae) from the abyssal manganese nodule field (Clarion-Clipperton Fracture Zone, North-Eastern Tropical Pacific). Helgoland Marine Research 66: 413–462. DOI:10.1007/s10152-011-0282-z.

13. KIHARA, T.C. & MARTÍNEZ ARBIZU, P. (2012): Three new species of Cerviniella Smirnov, 1946 (Copepoda: Harpacticoida) from the Arctic. Zootaxa 3345: 1-33. DOI: 10.5281/zenodo.281475

12. KIENEKE, A. & OSTMANN, A. (2012): Structure, function and evolution of somatic musculature in Dasydytidae (Paucitubulatina, Gastrotricha). Zoomorphology 131: 95-114.

11. MENZEL, L. (2011): First descriptions of copepodid stages, sexual dimorphism and intraspecific variability of Mesocletodes Sars, 1909 (Copepoda, Harpacticoida, Argestidae), including the description of a new species with broad abyssal distribution. Zookeys 96: 39-80. doi:doi:10.3897/zookeys.96.1496

10. VEIT-KÖHLER, G., GUILINI, K., PEEKEN, I., SACHS, O., SAUTER, E.J. & WÜRZBERG, L. (2011): Antarctic deep-sea meiofauna and bacteria react to the deposition of particulate organic matter after a phytoplankton bloom. Deep-Sea Research II 58: 1983–1995. doi: 10.1016/j.dsr2.2011.05.008

09. WULFKEN, D., E.F. WILTS, P. MARTÍNEZ ARBIZU, & W.H. AHRLICHS. (2010): Comparative analysis of the mastax musculature of the rotifer species Pleurotrocha petromyzon (Notommatidae) and Proales tillyensis (Proalidae) with notes on the virgate mastax type. Zoologischer Anzeiger 249(3–4): 181–194.

08. WILTS, E.F. & W.H. AHLRICHS. (2010): Proales tillyensis sp.n. (Monogononta: Proalidae), a new rotifer species from North-West Germany, with reconstruction of its somatic musculature. Invertebrate Zoology 7(1): 29–46.

07. WILTS, E.F., WULFKEN, D. & AHLRICHS, W.H. (2010): Combining confocal laser scanning and transmission electron microscopy for revealing the mastax musculature in Bryceella stylata (Milne, 1886) (Rotifera: Monogononta). Zoologischer Anzeiger 248(4): 285–298.

06. SCHULZ, M. & GEORGE, K.H. (2010): Ancorabolus chironi sp. nov., the first record of a member of the Ancorabolus-group (Copepoda: Harpacticoida: Ancorabolidae) from the Mediterranean. Marine Biodiversity 40: 79-93

05. MICHELS, J. & BÜNTZOW, M. (2010): Assessment of Congo red as a fluorescence marker for the exoskeleton of small crustaceans and the cuticle of polychaetes. Journal of Microscopy 238: 95-101.

04. WILTS, E.F., AHLRICHS ,W.H. & MARTÍNEZ ARBIZU, P. (2009): The somatic musculature of Bryceella stylata (Milne, 1886) (Rotifera: Proalidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology. Zoologischer Anzeiger 248(3): 161–175.

03. RIEMANN, O., WILTS, E.F., AHLRICHS, W.H. & KIENEKE, A. (2009): Body musculature of Beauchampiella eudactylota (Gosse, 1886) (Rotifera: Euchlanidae) with additional new data on its trophi and overall morphology. Acta Zoologica 90: 265-274.

02. RIEMANN, O., MARTÍNEZ ARBIZU, P. & KIENEKE, A. (2008): Organisation of body musculature in Encentrum mucronatum Wulfert, 1936, Dicranophorus forcipatus (O.F. Müller, 1786) and in the ground pattern of Ploima (Rotifera: Monogononta). Zoologischer Anzeiger 247: 133-145.

01. KIENEKE, A., MARTÍNEZ ARBIZU, P. & RIEMANN, O. (2008): Body Musculature of Stylochaeta scirtetica Brunson, 1950 and Dasydytes (Setodytes) tongiorgii (Balsamo, 1982) (Gastrotricha: Dasydytidae): A Functional Approach. Zoologischer Anzeiger 247: 147-158.