CONFOCAL MICROSCOPY LABORATORY (MK)

Personnel
Head of Laboratory: Teresa Kasprzycka-Guttman, Professor.
tel.: 851 52 41, 851 43 81 w. 319,
Irena Misiewicz; M.Sc., tel. 851 52 41, 851 43 81 w. 319, 
Katarzyna Skupińska; Skupiska.Sc. tel. 851 52 41, 851 43 81 w. 319,

Activities:
Quality Control of medicinal products:

  • Microscopic determination of insulin crystal shape. 

Quality assurance:

  • Introducing Quality System according to  PN-EN ISO/IEC 17025:2001;
  • Elaborating of the documents required to obtain accreditation of the Laboratory

Research:

  • Synthesis and research on isothiocyanates (sulforaphane and its analogs). These compounds exhibit chemopreventive properties and are found mainly in broccoli and all vegetables of Brassica family. These compounds are extracting from natural substances and analogs are syntethised. The studies are conducted on cancer cell lines and limphoblastoids derived from healthy humans and from humans of increased cancer risk.
  • Study on policycylic aromatic hydrocarbons (PAH). These compounds are considered carcinogenic. Our aim is to investigate mechanism and to find if sulforaphane can block the carcinogenesis induced by PAHs. In particular inhibitory effect of sulforaphane on creation of adduct of PAHs metabolites and protein or DNA. 
  • Study on anticarcinogenic properties of Selol. The aim of study is to evaluate  antitumor activity of SELOL by in vitro screening studies using various tumor cell lines. Also mechanism of antitumor activity of SELOL are studied by means of confocal microscopy.
  • Confocal microscopic investigation of cell entering and intracellular distribution and of aminoacid derivatives of porfirynes used in dynamic phototherapy.

Educational activity:

  • Training in the fields of: cell maintenance, preparation of microscopic slides especially for purpose of confocal microscopy ( selection of dyes, microscopic slides), observation with confocal microscopy and image analysis
  • Conducting and supervising over the M.Sc. thesis

Cooperation

  • Our laboratory obtains study objects from Pomeranian Medical University, professor J. Lubinski: cells derived from humans with increased cancer risk. The effect of sulforaphane is investigated on human limfoblastoid derived from lymphocytes B immortalized with Epstein-Barr virus obtained from patients with hereditary BRCA-1 gene mutation. Sulforaphane cytotoxity, induction of apoptosis, induction of chemoprevention markers are studied as well as quinine reductase activity - the enzyme rised as a marker of xobiotics II phase metabolism and the level of glutathione. Research is conducted and the results are published in cooperation.
  • The measurements are done for the NIL's Departments and Laboratories, e.g. Biochemistry and Biopharmaceuticals Department.
  • Department of Chemistry of Warsaw University and Medical University of Warsaw - students training (also Master Thesis)
  • Military University of Technology - investigation of aminoacid derivatives of porfirynes entering into cells.

Services:

  • Sample preparation
  • Sample staining
  • Microscopic image recording
  • Image analysis

In particular:

  • Cytotoxic and chemopreventive properties of natural and synthetic substances.
  • Study on drugs of crystal shape.
  • Influence of shampoo, conditioners and hair dyes on hair quality.
  • Cytotoxity of natural and synthetic substances on multi-drug resistant cells.
  • Tooth surface analysis (enamel quality, the structure of tooth surface and quality of filling).

Equipment:

  • Invert microscope Olympus IX70

(with epifluorescence, phase contrast, Nomarski contrast)

  • Scanning unit Fluoview 500:
  • 4 channels of fluorescence detection
  • Transmitted light
    • Lasers:
      • Ar 351 nm
      • Ar 488 nm 
      • He-Ne 543 nm
      • He-Ne 633 nm
  • Scanning in xy xz surfaces
  • 3D reconstruction
  • Time measurements: xyt, xyzt
  • Laminar Flow Hood
  • Incubator CO2
  • incubator, pH-meter, centrifuge, analytical balance, steam autoclave, WesternBlott set

Publications since 2002

  • Misiewicz I, Kozar A, Skupinska K, Kowalska E, Lubinski J and Kasprzycka -   Guttman T: Inhibition of cell cycle and induction of apoptosis by sulforaphane in cell lines carrying various inherited BRCA1 mutations. Oncol Rep, 2005; 13(4): 659-65
  • K. Skupińska, I. Misiewicz, T. Kasprzycka-Guttman: "Polycyclic aromatic hydrocarbons: physicochemical properties, environmental appearance and impact on living organisms" Acta Poloniae Pharmaceutica -Drug Research 61: 233-240, 2004           
  • Misiewicz I., Skupińska K. and Kasprzycka-Guttman T .: "Sulforaphane-mediated induction of phase 2 detoxifying enzyme NAD(P)H: quinone reductase and apoptosis in human lymphoblastoid cells" Acta Biochim P ol, 2004; 51(3): 711-721
  • Misiewicz I., Skupińska K. and Kasprzycka-Guttman T.: "Sulforaphane and 2-oxohexyl isothiocyanate induce cell growth arrest and apoptosis in L-1210 leukemia and ME-18 melanoma cells" Oncology Reports, 2003 Nov-Dec; 10(6):2045-50.
  • Misiewicz I., Skupinska K. And Kasprzycka-Guttman T.: "Effect of sulforaphane on human lymphoblastoid cells", Amino Acids 25 (2) p. 148, 2003
  • Skupińska K, Bojarska E,  Misiewicz I, Kasprzycka-Guttman T. "Determination of the Affinity of Polycyclic Aromatic Hydrocarbons toward Serum Albumin.", Amino Acids 25 (2) p. 148, 2003
  • Fijałek Z., Sarna K. and Kasprzycka-Guttman T.: "Voltammetric and electrochemical quartz crystal microbalance studies of sulforaphane and its Zn(II) complexes" J. Pharm. Biomed. Anal. 32 (2003) 967-974
  • Treszczanowicz T., Kasprzycka-Guttman T. and Treszczanowicz AJ: "Solubility of b-Carotene in Binary Solvents Formed by some Hydrocarbons with Dibutyl Ether and 1,2-Dimethoxyethane" J Chem Eng Data; 2003; 48(6) 1517-1520

4 i 5 -presentation at 8th International Congress on Amino Acids and Protein, Roma 5-9 September 2003.

Polish Patent application:
1. New compounds, analogues of sulforaphane and pharmaceutical agent.