Laboratory of Flow Cytometry (CP)

Personnel:
Head of the laboratory: Janusz S. Skierski, M.D., Ph.D.
phone 851-43-79,    841-06-42 ext. 175, fax: 841-25-09, room 412, e-mail:
Secretary: Danuta Makarewicz
phone 851-43-79, 841-06-42 ext. 420, fax: 841-25-09, room 412, e-mail:

Deputy Head: Miros³awa Koronkiewicz; M. Sc., phone 851-43-79, 841-06-42 ext. 450, room 450, e-mail: Sylwia Szafraniec, M.D.; phone. 851-43-79, 841-06-42 ext. 450, room 450; e-mail: Renata Krzysztofik, technician; phone 851-43-79, 841-06-42 ext. 420, room 420

Activities:

Quality control of medicinal products:
Those activities concentrate on determination of cytotoxicity of various chemical compounds in vitro and also on human immunology (determination of lymphocyte subpopulation in the blood). Determination of various cell constituents (esp. DNA), labeled with fluorescent dyes, using flow cytometry is performed as well as the determination of apoptotic cell fraction, especially in the case of the neoplastic cells and evaluation of cell cycle parameters and the viability tests on cells exposed to various cytotoxic and cytostatic chemicals.

Quality Assurance:
Quality assurance system according to PN EN ISO/IEC 17025:2001 is being implemented. Tests are performed according to procedures and instructions. Results are monitored by certified reference materials and the Lab actively participates in international and domestic QA programs. All equipment is metrologically controlled. Biological tests are performed in  controlled conditions. Premises are subjected to microbiological purity tests.

Research:

Research activity concentrates on biology of cancer cell. Mechanisms of action of anticancer substances (existing as well as potential drugs) as well as the influence of these substances on the cell cycle parameters and the induction of apoptosis are tested.
The following techniques are used:

  • flow cytometry,
  • microscopy -  fluorescence, phase contrast and Nomarsky contrast,
  • UV- Vis spectrophotometry,
  • molecular biology techniques.

Various neoplastic and non-neoplastic cell lines are used to perform in vitro tests. No tests on animals are performed.

Research Projects:
We are involved in grants from the Ministry of Science and Information Society Technologies.
4 P05F 014 19 "Elaboration of strategy of designing drugs active against multi-drug resistant cancers"
3 P05F 017 22 "Evaluation of activity of  new anthracycline derivatives as potential drugs active against multi-drug resistant cancers". This grant is realized in collaboration with professor W. Priebe from MD Anderson Cancer Center (Houston, TX, USA).
3 P05F 010 23 "Genistein derivatives - new drugs with anticancer activity".

Educational activity:
Organization and conducting courses in the field of flow cytometry and fluorescent microscopy.

Cooperation:

National:
Flow Cytometry Lab works in a close cooperation with many of academic schools and research institutes:

  • Medical University of Warsaw,
  • Warsaw Agricultural University (SGGW),
  • NENCKI Institute of Experimental Biology PAS,
  • Medical Research Center, PAS,
  • Institute of Psychiatry and Neurology in Warsaw,
  • Institute of Oncology in Warsaw,
  • Institute of Hematology  in Warsaw and others.

International:

  • Cancer Research Institute at The New York Medical College, Hawthorne, NY, USA,
  • MD Anderson Cancer Center, Houston, TX, USA.

The Lab is a leading laboratory in Poland in the field of flow cytometry. All persons are the active members of Polish Society of Cytometry.
Dr J. Skierski is active also in the IV Local Ethical Committee for the Experiments on Animals located in NIPH.

Services:

  • Determination of DNA content in the cells, together with the determination of cell cycle parameters in clinical and experimental samples.
  • Determination of apoptotic cell fraction, using the Annexin V/Pi, PARP cleavage and TUNEL methods.
  • Various viability tests.
  • Determination of LD50 value using cell lines in culture.
  • Immunophenotyping of human blood lymphocytes.
  • Cell morphology using fluorescence microscopy, phase-contrast and Nomarsky contrast techniques.
  • High resolution digital microphotography (5 Mpixel).

 

Equipment:

  • FACS Vantage flow cytometer (sorter and analyzer). Excitation wavelength: UV (351 nm), blue (488 nm), red (633 nm). Analysis channels: blue (424 nm), green (530 nm), orange (575 nm), red (630 nm), far red (>660 nm).
  • FACS Calibur flow cytometer (analyzer). Blue argon laser (488 nm) as an excitation light source, fluorescence channels green, orange and red.
  • Olympus BX60 fluorescence microscope with full optics and DP50 digital camera (cooled CCD, 5 Mixels).
  • Shimadzu UV-1700 PharmaSpec spectrophotometer (190 - 1100 nm).
  • PowerWave XS Biotek microplate reader.
  • Tissue culture and cell bank equipment (laminar flow, CO2 incubator, liquid nitrogen tanks (dewars), inverted microscope (Olympus) etc).