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[compost_tea] Re: Observing Microbial Biofilms on Leaf Surfaces + Isolation of Culturable Micr
Hi Bob -
SFI uses fluorescein diacetate (FDA) in combination with
epifluorescent microscopy to measure active microbial
biomass, and a range of microscopy methods for all the
different parameters they measure. Elaine can provide exact
Elaine's methodologies were developd through graduate and
post-graduate research going back to early 1980s.
See a bibliography of her papers at:
Sunburst Nominees with SFI | Australia
Ingham, E. R., and D. A. Klein. 1984. Soil fungi: relationships
between hyphal activity and staining with fluorescein
diacetate. Soil Biology and Biochemistry 16:273-278.
Ingham, E.R. and D.A. Klein. (1984). Soil fungi: Measurement
of hyphal length. Soil Biol. Biochem. 16:279-280.
Stamatiadis, S., J.W. Doran and E.R. Ingham. (1990). Use of
staining and inhibitors to separate fungal and bacterial
activity in soil. Soil Biol. Biochem. 22:81-88.
Mary Appelhof published an article in WormEzine (August 2003)
that summarized the SFI lab techniques:
"Another capability that Dr. Ingham has is to determine which
cells are active at the time the lab receives the samples.
Many bacteria, protozoa, and fungi would be in a dormant
or inactive state. Others are actively metabolizing at the
time they make the preparation. By adding fluorescein diacetate
(FDA) to the preparations, any cell whose mitochondria are
active take up the FDA. When the technicians look at the sample
using ultraviolet light with the microscope, the active cells
show up a bright fluorescent yellow/green. By using a dark
field on the microscope the bright yellow-green cells show up
very readily, enabling the viewer to ascertain the number
of active bacteria, fungi, or protozoa as compared with the
total numbers or hyphal length."
Etc... keep reading, this article provides an excellent
summary of SFI procedures.
WormEzine Vol. 2, No. 7
FDA and acridine orange (used in the French study) are
fluorochromes. There's a whole bunch of the fluorochromes.
Nikon Microscopy: Yellow Fluorescent Protein (YFP) Filter Set
Fluorochromes and recommended filter pairs
--- In firstname.lastname@example.org, "bobkacalek" <bob@s...> wrote:
> Hi Steve,
> Thanks for sharing, very interesting.
> The advantage of quality ACT is the abundance of diversity in Micro-
> organisms delivered vs one or two specific cultures.
> Are the methods described (Epifluorescence microscopy, scanning
> electron microscopy, and confocal laser scanning microscopy were
> to observe microbial biofilms directly on leaf surfaces)similar to
> methods used by SFI? Maybe Dr. E will respond?
> Thanks Again,
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