Chapter Outline: Assessment of prokaryotic
biological activity, at the single cell level, by combining
microautoradiography with fluorescence in
situ hybridization (FISH)
Proposed title: Assessment of prokaryotic
biological activity, at the single cell level, by combining
microautoradiography and fluorescence in
situ hybridization (FISH)
Chapter no.: _________
Author(s): Cleber C. Ouverney (PhD)
Department
of Biological Sciences
San
Jose State University
San
Jose, CA 95192-0100
Phone: +1(408) 924-4806
Fax: +1(408) 924-4840
E-mail:
cleber.ouverney@sjsu.edu
Notes:
· Some materials in this chapter may overlay with those covered in the Molecular Detection of Target Organisms and Sequences as well as chapters covering Physiological Detection of microorganisms.
Proposed topics
1) A culture-independent
method to render function of the 99% microbes in most environmental sites.
2) A brief overview of in situ detection of biological activity
by environmental microbes.
3) How do MAR-FISH,
STARFISH, Micro-FISH methods work to assess microbial metabolic activity in situ at the single cell level.
4) Advantages to the
approach, specifically the ability to simultaneously detect specific microbes
using fluorescently labeled oligonucleotide probes and detect the capacity of
microbes to uptake specific dissolved nutrient.
5) What are the pros and
contras of such approach?
6) How do the
microautoradiography-FISH methods compare to similar approaches?
7) What substrates are used
to render what type of metabolic activity?
8) Suggestions on how to
increase fluorescence signal to detect slow growing organisms.
9) Will such laborious
methods withstand the era of fast genomic sequencing?
10) Broad implications from
studies applying such methods in the understanding of biogeochemical cycling of
dissolved organic and inorganic nutrients in aquatic systems.
11) What other discoveries in
the microbial world have been made from these techniques?
Chapter Highlights
The
following concepts will be conveyed in this chapter:
1.
How metabolic activity can be assessed from uncultured microbes in natural
microbial communities.
2.
Is Autoradiography-FISH technique right for my application? What do I need to
know to make an intelligent decision?
3.
Can genomic sequence data be tested/validated using this approach? How?
