Confocal Microscopy Laboratory

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Confocal Microscopy Laboratory

Laboratory provides research services that are focused primarily on analysis of biological processes in living cells and tissues using fluorescence microscopy. It is equipped with confocal microscopes, which are well suited for 3D analysis of stained specimens and live imaging of cells and tissues in vivo or in culture. Research activities in the laboratory concentrate on how cellular physiology is affected by various conditions and are aimed at discovery and development of new bio-active substances and diagnostic tests.

MAIN SERVICES:

High-resolution fluorescence analysis of cultured cells and tissue sections

morphometry, evaluation of intracellular distribution of structures of interest, 3D reconstruction, co-localization assays

Live microscopic analysis of cells and tissues in vivo and in culture

longitudinal analysis of morphology, migration studies of cells and intracellular structures in real time in controlled environment (temperature, humidity, CO2 and O2), production of reactive oxygen species

Microscopic high-throughput screening

Qualitative and quantitative analysis of microscopic images

SPINNING DISK CONFOCAL MICROSCOPE ZEISS CELL OBSERVER SD

  • Inverted stand
  • Spinning disk module Yokogawa CSU-X1A 5000
  • Lasers: 405, 488, 561, 639 nm
  • Dual camera setup: EMCCD QImaging Rolera EM-C2
  • Objectives: 10x (NA 0.30), 20x (NA 0.4 LD), 25x (NA 0.80 W), 40x (NA 0.6 LD; NA 1.20 W; NA 1.4 Oil), 63x (NA 1.20 W; NA 1.4 Oil), 100x (NA 1.46 Oil)
  • Motorized stage with Definite Focus
  • Incubator XLmulti S1 and heating insert for 96-well plates
  • Temperature and O2/CO2 controller
  • DirectFRAP module for FRAP (Fluorescence Recovery After Photobleaching)

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LASER SCANNING CONFOCAL MICROSCOPE LEICA SP8 MP

  • Upright stand
  • Conventional and resonant (8 kHz) scanner
  • VIS lasers: 488, 552, 638 nm
  • IR laser: femtosecond pulsed Chameleon Vision II, tuning range: 680-1080 nm
  • Detectors: 3 internal PMT, 4 external NDD-PMT
  • Objectives: 10x (NA 0,40), 20x (NA 0,75), 25x (NA 0,95 W), 40x (NA 1,10 W; NA 1,3 Oil), 63x (NA 1,4 Oil)
  • Motorized stage
  • Temperature controller and heating insert for Petri dishes
  • Set-up for intravital imaging of mice

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IMAGE ANALYSIS SOFTWARE

  • Imaris 8 (Bitplane)
  • Huygens Professional (SVI)
  • Image-Pro Premier (Media Cybernetics)

Grzegorz Chodaczek, PhD – Head of the Laboratory
grzegorz.chodaczek@eitplus.pl

Katarzyna Mikołajewicz, M.Sc. – PhD student
katarzyna.mikolajewicz@eitplus.pl

Monika Toporkiewicz, PhD – Post-doc
monika.toporkiewicz@eitplus.pl

  1. Drabik D, Przybyło M, Chodaczek G, Iglič A, Langner M. The modified fluorescence based vesicle fluctuation spectroscopy technique for determination of lipid bilayer bending properties. Biochim Biophys Acta. 2015 Nov 23;1858(2):244-252.
  2. Shaked I, Hanna RN, Shaked H, Chodaczek G, Nowyhed HN, Tweet G, Tacke R, Basat AB, Mikulski Z, Togher S, Miller J, Blatchley A, Salek-Ardakani S, Darvas M, Kaikkonen MU, Thomas GD, Lai-Wing-Sun S, Rezk A, Bar-Or A, Glass CK, Bandukwala H, Hedrick CC. Transcription factor Nr4a1 couples sympathetic and inflammatory cues in CNS-recruited macrophages to limit neuroinflammation. Nat Immunol. 2015; 16(12):1228-34.
  3. Hanna RN, Cekic C, Sag D, Tacke R, Thomas GD, Nowyhed H, Herrley E, Rasquinha N, McArdle S, Wu R, Peluso E, Metzger D, Ichinose H, Shaked I, Chodaczek G, Biswas SK, Hedrick CC. Patrolling monocytes control tumor metastasis to the lung. Science. 2015; 350(6263):985-90.
  4. McArdle S, Chodaczek G, Ray N, Ley K. Intravital live cell triggered imaging system reveals monocyte patrolling and macrophage migration in atherosclerotic arteries. J Biomed Opt. 2015; 20(2):26005.
  5. Mikulski Z, Johnson R, Shaked I, Kim G, Nowyhed H, Goodman W, Chodaczek G, Pizarro TT, Cominelli F, Ley K. SAMP1/YitFc mice develop ileitis via loss of CCL21 and defects in dendritic cell migration. Gastroenterology 2015; 148(4):783-793.e5.
  6. Vicente-Suarez I, Larange A, Reardon C, Matho M, Feau S, Chodaczek G, Park Y, Obata Y, Gold R, Wang-Zhu Y, Lena C, Zajonc DM, Schoenberger SP, Kronenberg M, Cheroutre H. Unique lamina propria stromal cells imprint the functional phenotype of mucosal dendritic cells. Mucosal Immunol. 2015; 8(1):141-51.
  7. Verma S, Wang Q, Chodaczek G, Benedict CA. Lymphoid tissue stromal cells coordinate innate defense to cytomegalovirus. J Virol. 2013; 87(11):6201-10.
  8. Ariotti S, Beltman JB, Chodaczek G, Hoekstra ME, van Beek AE, Gomez-Eerland R, Ritsma L, van Rheenen J, Marée AF, Zal T, de Boer RJ, Haanen JB, Schumacher TN. Tissue-resident memory CD8+ T cells continuously patrol skin epithelia to quickly recognize local antigen. Proc Natl Acad Sci USA. 2012; 109(48):19739-44.
  9. Koltsova EK, Garcia Z, Chodaczek G, Landau M, McArdle S, Scott SR, von Vietinghoff S, Galkina E, Miller YI, Acton ST, Ley K. Dynamic T cell-APC interactions sustain chronic inflammation in atherosclerosis. J Clin Invest. 2012; 122(9):3114-26.
  10. Chodaczek G, Papanna V, Zal MA, Zal T. Body-barrier surveillance by epidermal γδ Nat Immunol. 2012; 13(3):272-82.
  11. Zal T, Chodaczek G. Intravital imaging of anti-tumor immune response and the tumor microenvironment. Semin Immunopathol. 2010; 32(3):305-17.
  12. Chodaczek G, Bacsi A, Dharajiya N, Sur S, Hazra TK, Boldogh I. Ragweed pollen-mediated IgE-independent release of biogenic amines from mast cells via induction of mitochondrial dysfunction. Mol Immunol. 2009; 46(13):2505-14.
  13. Chodaczek G, Zimecki M, Lukasiewicz J, Lugowski C. Lactoferrin-monophosphoryl lipid a complex enhances immunity of mice to Plesiomonas shigelloides CNCTC 138/92. Acta Biochim Pol. 2008; 55(1):91-6.
  14. Bacsi A, Chodaczek G, Hazra TK, Konkel D, Boldogh I. Increased ROS generation in subsets of OGG1 knockout fibroblast cells. Mech Ageing Dev. 2007; 128(11-12):637-49.
  15. Zimecki M, Kocieba M, Chodaczek G, Houszka M, Kruzel M. Lactoferrin Ameliorates Clinical Symptoms of Experimental Encephalomyelitis in Lewis Rats. J Neuroimmunol, 2007; 182(1-2):160-6
  16. Chodaczek G, Zimecki M, Lukasiewicz J, Lugowski C. A complex of lactoferrin with monophosphoryl lipid A is an efficient adjuvant of the humoral and cellular immune response in mice. Med Microbiol Immunol (Berl). 2006; 195(4):207-16.

OPUS 8: Constitutive autoreactivity of gamma-delta T cells as a beneficial mechanism of epidermal barrier surveillance
project leader – Grzegorz Chodaczek, grant from National Science Centre, Poland, funding period 09.2015-09.2018

SONATA BIS 4: Multiparameter characterization of surveillance of reproductive epithelium homeostasis by gamma-delta T cells that includes high resolution intravital microscopy
project leader – Grzegorz Chodaczek, grant from National Science Centre, Poland, funding period 07.2015-07.2020

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Aleksandra Borek

Key Account Manager

Aleksandra Borek

tel: 510 131 925

aleksandra.borek@eitplus.pl

Posted by Agata Kołacz, Posted on 08.10.2015
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