METHODOLOGIES FOR THE STUDY OF FILAMENTOUS CYANOBACTERIA BY FLOW CYTOMETRY

• Type: Article in International Scientific Journal
• Date: 2012

Authors

Azevedo, R (Author) Other; Rodriguez, E (Author) Other; Figueiredo, D (Author) Other; Peixoto, F (Author) Other; Santos, C (Author) Other

Journal

Title: Fresenius Environmental Bulletin

Vol. 21 No. 8

Pages: 679-684

ISSN: 1018-4619

Publisher: Technische Universitat Munchen

Indexing

ISI Web of Knowledge - 2 Citations

Scopus - 4 Citations

Other information

Authenticus ID: P-002-GKF

Abstract (EN): Flow cytometry (FCM) is largely used in phytoplankton analyses, but its potential for analysing filamentous algae is limited by the difficulty to obtain individualised cells. To overcome this, we developed a rapid and low-cost method for isolating high yields of cells from filamentous cyanobacteria (Anabaena cylindrica, Anabaena cf. solitaria., Nostoc sp., Aphanizomenon gracile, Aphanizomenon sp. and Aphanizomenon flos-aquae) allowing maximum efficiency in cell isolation while minimizing degradation. In order to achieve an optimal methodology, six isolation solutions [Otto, Sgorbati and WPB buffers and FCB1 (0.1 % Triton-X100), FCB2 (0.2 % Triton-X100) and FCB5 (0.5 % Triton-X100) solutions], different incubation time and sonication periods were tested. Results indicate that cell isolation yield was mostly dependent on the isolation solution used. Among the buffers assayed, fluorescence microscopy observation revealed that WPB and FCB5 provided the highest yield of isolated cells. However, A. cylindrica and Nostoc sp. were more resistant and required sonication prior to incubation in WPB and FCB5 in order to obtain acceptable yields. In all species, FCB5 buffer provided best global results for cell integrity. The volume, granularity and autofluorescence of isolated cells were then characterized by FCM. In conclusion, this is the first report of an efficient, easy and low cost methodology for individualization of filamentous cyanobacteria cells, which enables their analysis by FCM, and will open a vast set of possibilities in this area of research.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 6