Fluorescence Microscopy

Rhizosphere and the Phyllosphere
Rhizosphere Introduction
Rhizospher Effect
Nitrogen Fixation in the Rhizosphere
Alteration of Rhizosphere Microflora
Associative and Antagonistic Activities in the Rhizosphere
Root Exudates
Organic Compounds Detected in Plant Root Exudates
Fungistasis
Techniques of Rhizosphere
The Soil Dilution and Plate Count Method
The Soil Plate
The Soil Box
The Contact Slide Technique
Artificial Rhizosphere
Model System

Fluorescence Microscopy

Fluorescence Microscopy

Suitable dilutions of soil suspensions are mixed with 1% Difco Bactoagar solutions and appropriate aliquots (0. 01 ml) transferred to microscopic glass slides by means of micropipettes. The glass slides may be etched suitably so that required amounts of the agar suspension get evenly and firmly distributed on the surface.

The slides are dried uniformly and stained for 2 minutes in a very dilute solution of acridine orange, care being taken to remove excess of the stain by dipping in water.

Acridine orange stain reacts with deoxyribonucleic acid of living bacteria and fluoresce red under a fluorescence microscope. Dead bacteria show greenfluorescence upon staining, a characteristic feature also shared by some Gram-negative bacteria which in fact has posed difficulties in differentiating living bacteria from dead ones by the use of this technique. Other factors such as the concentration of the dye, the time of exposure and pH of the medium govern the efficacy of this technique.

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