Post-congress Workshop on Practical Flow Cytometry

SCOPE:

The last two decades have seen a significant increase in the use of flow cytometry for estimation of genome size in plants (either in absolute terms or in relative units, as an indicator of ploidy level), with the data being successfully utilized in various fields of plant sciences, including biosystematics, ecology, evolutionary biology and biotechnology. The aim of the workshop is to present the best practice methodology for the genome size estimation and to alert to potential problems accompanying the analysis of plant samples. The theoretical part will focus on the biological significance of genome size variation in plants and on the methodological approaches used to analyse the nuclear DNA content of cells. A set of best practice rules will be presented, and special attention will be given to the use of flow cytometric results in plant biosystematics, ecology, and population biology. The practical session will include different protocols used for isolating and staining intact plant nuclei (both easy and challenging plant samples will be analyzed). The workshop is aimed at newcomers to plant flow cytometry as well as at more experienced users interested to absorb new knowledge.

SYLLABUS:

Theoretical part

• advantages and limitations of flow cytometry
• specific features of plant material
• DNA-selective fluorochromes, isolation and staining buffers
• standardization and DNA standards
• results quality
• secondary metabolites and their impact on quality of FCM measurements
• overview of best practice rules for genome size estimation
• basic references + web databases
• the use of flow cytometric data in plant biosystematics, ecology, and population biology (resolving complex low-level taxonomies, heteroploid and homoploid hybridization, cytogeography, breeding mode detection)

Practical part

• adjustment of flow cytometers
• estimation of genome size in leaf tissue using one-step protocol
• estimation of genome size in leaf tissue using two-step protocol
• estimation of genome size in mature seeds
• estimation of genome size in recalcitrant plant species
• interpretation of challenging histograms

LOCATION:

Laboratory of Flow Cytometry, Institute of Botany, Průhonice (lab web page)

TUTORS:

The team of the Laboratory of Flow Cytometry:

Jan Suda (personal web page)
Pavel Trávníček (personal web page)
Filip Kolář
Magdalena Lučanová
Jana Krejčíková
Tomáš Urfus
Petr Vít

Special guest: Matthias Steinberg (Partec GmbH)

INSTRUMENTS:

Partec PA II and Partec CyFlow SL flow cytometers (for details see Partec's web pages)

DATE(S):

The first course is scheduled for Friday, May 11, 2012.
Other courses can follow on Saturday 12.5. / Sunday 13.5., depending on the participant's interest.
The number of participants is limited to 10 per course. The minimum number of participants is 4 per course.

WORKSHOP FEE: 1 200 Kč or 50 EURO (payable on the spot)

Handout material will be provided to each participant.

A copy of the book Flow Cytometry with Plant Cells - Analysis of Genes, chromosomes and Genomes (eds. J. Doležel, J, Greilhuber & J. Suda - for more details see publisher's annotation) can be provided on request at reduced price (125 €) (please indicate your interest in the registration e-mail)

Seeds of suitable reference standards can also be provided on request (please indicate your interest in the registration e-mail)

For workshop registration, please, send message to Jan Suda (e-mail).