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Applied and Environmental Microbiology, November 2003, p. 6856-6863, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6856-6863.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Methodology of Protistan Discovery: from rRNA Detection to Quality Scanning Electron Microscope Images{dagger}

Thorsten Stoeck,1 William H. Fowle,2 and Slava S. Epstein1,2*

Marine Science Center, Northeastern University, East Point, Nahant, Massachusetts 01908,1 Department of Biology, Northeastern University, Boston, Massachusetts 021152

Received 5 May 2003/ Accepted 1 August 2003

Each year, thousands of new protistan 18S rRNA sequences are detected in environmental samples. Many of these sequences are molecular signatures of new protistan species, classes, and/or kingdoms that have never been seen before. The main goal of this study was to enable visualization of these novel organisms and to conduct quality ultrastructural examination. We achieved this goal by modifying standard procedures for cell fixation, fluorescence in situ hybridization, and scanning electron microscopy (SEM) and by making these methodologies work in concert. As a result, the same individual cell can now be detected by 18S rRNA-targeted fluorochrome-labeled probes and then viewed by SEM to reveal its diagnostic morphological characteristics. The method was successfully tested on a wide range of protists (alveolates, stramenopiles, kinetoplastids, and cryptomonads). The new methodology thus opens a way for fine microscopy studies of many organisms previously known exclusively by their 18S rRNA sequences.

* Corresponding author. Mailing address: Northeastern University, Marine Science Center, East Point, Nahant, MA 01908. Phone: (617) 373-4048. Fax: (617) 373-3724. E-mail: s.epstein{at}neu.edu.

{dagger} This is contribution 248 of the Marine Science Center of Northeastern University, Nahant, Mass.

Applied and Environmental Microbiology, November 2003, p. 6856-6863, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6856-6863.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kolodziej, K., Stoeck, T. (2007). Cellular Identification of a Novel Uncultured Marine Stramenopile (MAST-12 Clade) Small-Subunit rRNA Gene Sequence from a Norwegian Estuary by Use of Fluorescence In Situ Hybridization-Scanning Electron Microscopy. Appl. Environ. Microbiol. 73: 2718-2726 [Abstract] [Full Text]  
  • Stoeck, T., Schwarz, M. V. J., Boenigk, J., Schweikert, M., von der Heyden, S., Behnke, A. (2005). Cellular identity of an 18S rRNA gene sequence clade within the class Kinetoplastea: the novel genus Actuariola gen. nov. (Neobodonida) with description of the type species Actuariola framvarensis sp. nov.. Int. J. Syst. Evol. Microbiol. 55: 2623-2635 [Abstract] [Full Text]  


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