Avoiding autofluorescence problems: time-resolved fluorescence microscopy with plant and fungal cells in ectomycorrhiza
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Abstract
Sections of Pinus sylvestris-Suillus bovinus ectomycorrhiza were used to test the applicability of time-resolved fluorescence microscopy for studying plant and fungal material. The autofluorescence problems encountered with standard indirect immunofluorescence microscopy when studying specialized plant material were eliminated by the technique. The europium-labeled structures of both plant and fungal cells were the only visible areas in the sections. However, fine details were lost due to the coarse pixel size of the digital camera used to transfer the image on screen. Even with current resolution, time-resolved imaging can be an efficient method for detection of targets in both plant and fungal tissues with autofluorescence problems.
Key words
Autofluorescence Digital imaging Europium chelates Immunofluorescence microscopyPreview
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References
- Finlay RD, Read DJ (1986) The structure and function of the vegetative mycelium of ectomycorrhizal plants. II. The uptake and distribution of phosphorus by mycelial strands interconnecting host plants. New Phytol 103:157–165Google Scholar
- Mukkala V-M, Mikola H, Hemmilä I (1989) The synthesis and use of activated N-benzyl derivatives of diethylenetriaminetetraacetic acids: alternative reagents for labeling of antibodies with metal ions. Anal Biochem 176:319–325Google Scholar
- Sevéus L, Väisälä M, Syrjänen S, Sandberg M, Kuusisto A, Harju R, Salo J, Hemmilä I, Kojola H, Soini E (1992) Time-resolved fluorescence imaging of europium chelate label in immunohistochemistry and in situ hybridization. Cytometry 13:329–338Google Scholar
- Timonen S, Finlay RD, Söderström B, Raudaskoski M (1993) Identification of cytoskeletal components in pine ectomycorrhizas. New Phytol 124:83–92Google Scholar
- Väisälä M, Sevéus L, Kuusisto A, Harju R, Soini E (1990) Time-resolved fluorescence microscopy: elimination of autofluorescence in tissue specimens for image cytdmetry with fluorescent labeled probes. Micron Microscop Acta 21:178Google Scholar