Vrchní obrázek

International Society for Advancement of Cytometry

Live Education Delivery Task Force

invites you for

Prague Cytometry Workshop

April 21 – 24, 2016

 

 

 

NEWS


Dear colleagues,

Thank you for participating at the Prague cytometry workshop. I hope it was enjoyable and enriching for you as it was for us. I would be greatfull for any feedback so that we can keep what was valuable for you and improve what needs improvement. ISAC Live Education Task Force has agreed to support such a workshop every year in the Central and Eastern Europe. I would appreciate if you could help me to spread the word and suggest what people, institutions would be able (large lab with 3-4 three-laser cytometers) and willing (together with their staff) to host such event in the next years in our region. Also, watch our websites csac.cz / isac-net.org for future events and consider becoming a member of CSAC or ISAC. 

With best regards,

Tomas Kalina
On behalf of organizers


 

SCIENTIFIC PROGRAM

The preliminary program of the workshop, which consists of lectures and WetLabs is now availaible and can be download here: program.pdf
The participants will be split into two groups for the lectures (A and B) and 16 groups for the WetLabs. 
List of participants divided into specific groups for the lectures as well as the WetLabs can be found here: participants.pdf

LOCATION

The workshop will be held at 2 venues located just across the street.
CLIP: CLIP Laboratories, Department of Paediatric Hematology/Oncology, 2nd Medical School, Charles University Prague
Roentgenova street. More information: http://clip.lf2.cuni.cz/en/contact-us/
Map

HOMOLKA CONFERENCE CENTRE:
Na Homolce Hospital, Roentgenova street 2, Prague 5
Map
Entrance to the conference centre: https://goo.gl/maps/JxVc8ZjfWe12

How to get to HOMOLKA CONFERENCE CENTRE:

- 15 minutes walk from Nemocnice Motol Metro station (Green Line A) | map
- 2 minutes walk from "Sídliště Homolka” bus stop (bus line no. 167) | map

More information about Prague public transport (incl. connection search app) can be found at http://www.dpp.cz/en

Registration starts on Thursday at 12.00 in the foyer of the HOMOLKA CONFERENCE CENTRE.
Scientific program starts on Thursday at 13.00.


 

 

Dear colleagues,

 

ISAC and ČSAC invites you to "Prague Cytometry Workshop"

Workshop will cover 7 modules, 5 practical wet labs, and total duration of 3 days, up to 65 cytometrists from central Europe can particpate.


Workshop venue:
CLIP Laboratories, Department of Paediatric Hematology/Oncology, 2nd Medical School, Charles University Prague
http://clip.lf2.cuni.cz/en/contact-us/

 

Approximate timetable: 
Beginning on Thursday, April 21, 12.00
Conclusion on Sunday, April 24, 12.00

 

Program will include lectures for all participants and wet-lab modules in small groups (click for details):

Signal Transduction in Human Disease. D. Hedley, S.Chow

Cell Cycle, Proliferation and Apoptosis.  A. Krishan, K. Soucek, D. Hedley

Stem Cell Assays.  W. Telford, A. Krishan, K. Soucek

Basics of Flow Cytometry.  Z. Maciorowski, T. Kalina

Build your own cytometer, W. Telford, M. Schiemann

Multicolor module: compensation, staining and spillover P. Jain, Z. Maciorowski

Instrument maintenance, daily checks and modification. Ray Hicks

 

 

Organizers:
Awtar Krishan, Zosia Maciorowski and Tomas Kalina

 

Instruments used in wet-labs will include:
BD FACS Celesta (demo instrument kindly provided by BD), BC CytoFlex (demo instrument kindly provided by BC), Sony SP6800 spectral cytometer, BD FACS Aria III, BD FACS Canto II, BD LSR II

 

Registration:

Early registration fee by 15.3.2016: 4800 CZK (= 178 EUR) (VAT included)
LATE registration fee from 16.3.2016: 5500 CZK (= 204 EUR) (VAT included)

Participation fee includes coffee breaks, lunch box, social dinner and participation to one wet-lab. 

Each participant will be assigned to ONE wet-lab. Enter your preference during registration, assignment will be based on availability (maximum 5 participants per instrument in each wet-lab).

 

REGISTRATION IS CLOSED 

 

SOCIAL DINNER:

Sightseeing tour of Prague + Dinner

More information will be available soon.

 

POSSIBILITIES OF ACCOMMODATION:

Closest (15min walk) is a bed and breakfast Kotlarkawww.usedlost-kotlarka.cz

By 10-15 min bus ride (every 5 minutes):

Hotel Ibis | www.accorhotels.com/gb/hotel-5211-ibis-praha-mala-strana/index.shtml

Hotel Kavalirwww.hotelkavalirprague.cz

 

or any other hotel in the center (close to metro line C - can be reached in 30min),

Hotel DAP | www.daphotel.cz

good value for its price is Hotel DAP

 

Sponsors:

 

 

    

 

 

Faculty (Click for Bio sketch data):

David Hedley

University of Toronto and Princess Margaret Hospital, Toronto, Ontario, Canada

Sue Chow

University of Toronto and Princess Margaret Hospital, Toronto, Ontario, Canada

Bill Telford

Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA

 

Zosia Maciorowski

Curie Institute, Paris, France.

Awtar Krishan

Chair of the Live Education Task Force of

The International Society for Advancement of Cytometry (ISAC).

 

  

Paresh Jain

Scientific Advisor, BD Biosciences, India.

Matthias Schiemann

Institute for Medical Microbiology, Immunology and Hygiene – Technische Universität München, Germany

 

Ray Hicks

Flowcell, UK

 

Karel Souček

Department of Cytokinetics, Institute of Biophysics, Academy of Sciences, Brno, Czech Republic


   

Veronika Kanderová

CLIP Cytometry, Department of Pediatric Hematology and Oncology, Charles University, Prague, Czech Republic

 Tomáš Kalina

CLIP Cytometry, Department of Pediatric Hematology and Oncology, Charles University, Prague, Czech Republic

   

 

                      

Program details:

Build your own cytometer (LM-1B)

(Bill Telford)

Lecture –Build Your Own Cytometer.  One of the best ways to learn flow cytometry is to actually build one!  In this laboratory, we will build a working flow cytometer from individual components, using a laser, a flow cell, fluidics and optics to construct an actual instrument capable of analyzing microspheres and cells.  Students will be able to examine and install all the mechanical, optical and electronic components of a modern flow cytometer and see how they interact to form a working instrument.

Lab module- Stem Cell Assays.   In this lab Drs. Telford and Krishan will discuss different methods for the identification of stem cells by laser flow cytometry.



Intracellular Cytometry: Signaling and Cell Cycle Regulation (LM-15)

(David Hedley, Sue Chow, Veronika Kanderová)

The large majority of proteins involved in the regulation of cell signaling, survival, and growth regulation are intracellular. This lab covers the technical aspects of intracellular antigen staining for flow cytometry. It includes approaches to cell fixation and permeabilization that optimize intracellular labeling while preserving light scatter and phenotypic markers. Although more demanding than cell surface staining, mastery of intracellular cytometry enables the study of fundamental regulatory mechanisms of normal and abnormal cell biology, many of which remain under‐explored. In the second half of the lab we will illustrate this with (1) the activation of signal transduction pathways that regulate the acute inflammatory response via the NFkB transcription factor, and (2) molecular mechanisms that regulate movement through the cell division cycle.


Cell Cycle, Proliferation and Apoptosis.  (LM-2)

(Awtar Krishan, David Hedley, Karel Souček)

Flow cytometry is a rapid method for monitoring of cellular DNA content, cell cycle distribution and detect cells in proliferation and apoptosis.  In this wet lab, we will demonstrate flow cytometric methods for DNA content and cell cycle analysis and for the detection of proliferating and apoptotic cells. In this lab module, we will prepare a variety of cells for flow cytometric determination of DNA content by the propidium iodide/hypotonic citrate method.  We will discuss techniques for the study of proliferation and apoptosis by laser flow cytometry.   


Multicolor module: compensation, staining and spillover (MM)

(Paresh Jain, Zosia Maciorowski)

Lecture: Polychromatic flow cytometry (Jain and Maciorowski)

The last decade has seen a paradigm shift in instrument design of flow cytometers enabling analysis of 6, 8 or more colors simultaneously. Development of new fluorochromes and antibody conjugates provides a new level of choice and control in designing robust assays that can detect and monitor multiple analytes to produce a tremendous amount of data while conserving sample. Selecting the right fluorochromes for one’s experiment requires the knowledge of various characteristics of a fluorochrome such as its excitation/emission spectra, brightness, spectral spillover and stability. In this talk, we will provide an overview of these properties of the fluorochromes and how they can impact the data quality and interpretation. We will also cover fundamental aspects of antigen-antibody interaction.

Lab - Polychromatic Flow Cytometry (Jain and Maciorowski)

In this lab-based module, participants will practice how to rank a variety of routine and novel fluorochromes based on their brightness, perform basic compensation experiments and study the effect of spectral spillover related spread on experimental results. The module also includes a step-by-step demonstration of an antibody titration experiment.

Tutorial – Multicolor Panel design:  (Jain, Maciorowski and Kalina)

Looking for the expression of specific cellular proteins is the most common application of flow cytometry but with the proliferation of fluorochromes, designing a multicolour panel can be challenging. In this tutorial based module we will synthesize the knowledge of fluorochrome properties to create multicolor panels of increasing complexity.


Stem Cell Assays  (LM-10)

(Bill Telford, Awtar Krishan, Karel Soucek)

In this lab we will review recent advances in stem cell analysis by flow cytometry, including cell surface phenotype analysis for tumor stem cell markers and physiological characteristics such as ABC transporter activity using pump-specific efflux probes and aldehyde dehydrogenase activity.

Instrument maintenance, daily checks and modification (IM.)

(Ray Hicks) 

TBA

 

organized in collaboration with CSAC

 

 
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