Bioimaging Facility

 

 

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LSM 980 – with Airyscan 2

Purchased: 2018
Latest upgrade: January 2021 – Airy Scan, additional lasers
 

Responsible person: Dr. Martin Mistrik (martin.mistrik@upol.cz)

Technology: Laser scanning confocal microscope Zeiss LSM980

Special features: 

  • Airy scan (super-resolution, high sensitivity)
  • High power UV laser (405nm, 200mW)
  • Live cells imaging support (incubator, 40x water objective with automatic dispenser, hardware autofocus)
  • Incubator with cooling (special in-house developed upgrade of microscopic incubator allowing microscopic procedures between 16-45°C)

Established techniques: 

  • Standard confocal microscopy of fixed and live specimens (high resolution, high sensitivity)
  • Long-term live cells imaging (high resolution, high sensitivity)
  • Laser micro-irradiation (DNA damage induction, FRAP)
  • High content microscopy of fixed and live specimens (multiwell plates scanning and signal analysis)

Selected publications using the technology:

Aging (Albany NY). 2021 Dec 6;13(23):25004-25024. doi: 10.18632/aging.203744. Epub 2021 Dec 6.
Mol Oncol. 2021 Oct 10. doi: 10.1002/1878-0261.13114. Online ahead of print.
Nat Commun. 2021 Jan 29;12(1):713. doi: 10.1038/s41467-021-20989-9.

 

LSM 780 – with Elyra PS.1 module and 355nm UV laser

Purchased: 2018
Latest upgrade: January 2021 – Airy Scan, additional lasers 

Responsible person: Dr. Martin Mistrik (martin.mistrik@upol.cz)

Technology: Laser scanning confocal microscope Zeiss LSM780

Special features: 

  • Super-resolution modules (structured illumination - SIM, and photoactivated localization microscopy - PALM) 
  • UV laser (355nm, 60mW)
  • Live cells imaging support (incubator, hardware autofocus)

Established techniques: 

  • Standard confocal microscopy of fixed and live specimens (high resolution, super-resolution)
  • Long-term live cells imaging (high resolution, super-resolution)
  • Laser micro-irradiation (DNA damage induction, FRAP)

Seelected publications using the technology:

Nat Commun. 2021 Jan 29;12(1):713. doi: 10.1038/s41467-021-20989-9.
Cells. 2020 Feb 18;9(2):469. doi: 10.3390/cells9020469.
Oncogene. 2019 Oct;38(40):6711-6722. doi: 10.1038/s41388-019-0915-2. Epub 2019 Aug 7.
DNA Repair (Amst). 2019 Jun;78:114-127. doi: 10.1016/j.dnarep.2019.04.001. Epub 2019 Apr 6. Prostate. 2019 Mar;79(4):352-362. doi: 10.1002/pros.23741. Epub 2018 Nov 29.
Nature. 2017 Dec 14;552(7684):194-199. doi: 10.1038/nature25016. Epub 2017 Dec 6.
Nature. 2017 Dec 14;552(7684):194-199. doi: 10.1038/nature25016. Epub 2017 Dec 6.
Oncogene. 2017 Apr 27;36(17):2405-2422. doi: 10.1038/onc.2016.392. Epub 2016 Nov 14.
J Proteome Res. 2016 Dec 2;15(12):4505-4517. doi: 10.1021/acs.jproteome.6b00622. Epub 2016 Nov 9
Sci Rep. 2016 Jan 18;6:19567. doi: 10.1038/srep19567.
Nat Protoc. 2015 Aug;10(8):1248-63. doi: 10.1038/nprot.2015.083. Epub 2015 Jul 23.
Cell. 2014 Jul 31;158(3):633-46. doi: 10.1016/j.cell.2014.05.046.
Biol Reprod. 2014 Jun;90(6):124. doi: 10.1095/biolreprod.114.118497. Epub 2014 Apr 30.
Plant Physiol. 2014 May;165(1):129-48. doi: 10.1104/pp.114.238477. Epub 2014 Mar 31. 

 

Zeiss Cell Observer - with spinning disk confocal unit 

Purchased: 2011
Latest upgrade: January 2018 – Super-sensitive camera 

Responsible person: Dr. Martin Mistrik (martin.mistrik@upol.cz)

Technology: Zeiss laser-based fluorescent microscope with Yokogawa spinning disk confocal unit 

Special features: 

  • Fluorescent wide field and confocal imaging modes
  • Super-sensitive camera (95% QE)
  • High speed imaging 
  • Live cells imaging support (incubator with CO2)

Established techniques: 

  • Standard wide field and confocal microscopy of fixed and live specimens in four fluorescent channel (high resolution, high sensitivity, high speed)
  • Long-term live cells imaging (high resolution, high sensitivity, high speed)
  • High content microscopy of fixed and live specimens (multiwell plates scanning and signal analysis)

Selected publications using the technology:

Aging (Albany NY). 2021 Dec 6;13(23):25004-25024. doi: 10.18632/aging.203744. Epub 2021 Dec 6.
DNA Repair (Amst). 2019 Jun;78:114-127. doi: 10.1016/j.dnarep.2019.04.001. Epub 2019 Apr 6.
J Proteome Res. 2016 Dec 2;15(12):4505-4517. doi: 10.1021/acs.jproteome.6b00622. Epub 2016 Nov 9. 

 

ScanR automated microscopic system 

Purchased: 2008
Latest upgrade: 2016 – new software 

Responsible person: Dr. Martin Mistrik (martin.mistrik@upol.cz)

Technology: Olympus fluorescent microscope designed for high content and high throughput microscopy  

Special features: 

  • Fluorescent and transmission light modes
  • Live cells imaging support (incubator)
  • Hardware autofocus

Established techniques: 

  • Standard wide field microscopy of fixed and live specimens in six fluorescent channel
  • Long-term live cells imaging
  • High content and high throughput microscopy of fixed and live specimens (multiwell plates scanning, advanced signal analysis)

Selected publications using the technology:

Cell Death Dis. 2022 Mar 4;13(3):203. doi: 10.1038/s41419-022-04651-w.
Cell Death Differ. 2022 Mar;29(3):687-696. doi: 10.1038/s41418-021-00882-0. Epub 2021 Oct 5.
Nature. 2018 Jul;559(7713):279-284. doi: 10.1038/s41586-018-0261-5. Epub 2018 Jun 27.
Nature. 2017 Dec 14;552(7684):194-199. doi: 10.1038/nature25016. Epub 2017 Dec 6.
Cell Cycle. 2018;17(1):92-101. doi: 10.1080/15384101.2017.1403685. Epub 2017 Dec 10.
Oncogene. 2017 Apr 27;36(17):2405-2422. doi: 10.1038/onc.2016.392. Epub 2016 Nov 14.
Nature. 2015 May 28;521(7553):541-544. doi: 10.1038/nature14328. Epub 2015 Mar 23.
Cell Rep. 2015 Mar 17;10(10):1749-1757. doi: 10.1016/j.celrep.2015.02.028. Epub 2015 Mar 12.
Nat Commun. 2014 Nov 5;5:5348. doi: 10.1038/ncomms6348.
Cancer Res. 2011 Mar 15;71(6):2129-39. doi: 10.1158/0008-5472.CAN-10-2684.
J Cell Biol. 2009 Sep 7;186(5):655-63. doi: 10.1083/jcb.200812138. 

 

AFM Bruker Bioscope Catalyst 

Purchased: 2011
Latest upgrade: 

Responsible person: Dr. Hana Kolářová (hana.kolarova@upol.cz )

Technology: Olympus Atomic force microscope

Special features: 

  • High resolution imaging (nm)
  • Young's modulus (elastic modulus) determination
  • Measurement both in air and fluid
  • In combination with brightfield microscopy

Established techniques: 

  • AFM of biocomatible materials, fixed cells and tissues, bacteria, fibrils, nanoparticles
  • 2D and 3D mapping of the surface and determination of elastic properties

Selected publications using the technology:

Photodiagnosis and Photodynamic Therapy; 2021 34:102224. doi: 10.1016/j.pdpdt.2021.102224
Scientific Reports; 2021 11:6786. doi: 10.1038/s41598-021-85828-9
Cells Tissues Organs 2020 209(4-6):177-188. doi: 10.1159/000512807. Epub 2021 Feb 15.
Appl Immunohistochem Mol Morphol. 2020 Feb;28(2):166-174. doi: 10.1097/PAI.0000000000000689
Microscopy and Microanalysis 2020 26:139–147. doi:10.1017/S1431927619015113
Lekar a technika – Clinician and Technology 2020 50(3):114–121. doi: 10.14311/CTJ.2020.3.05
Lekar a technika – Clinician and Technology 2019 49(2):41-45. doi: 10.14311/CTJ.2019.2.01
Cells Tissues Organs 2019;208:37–47. doi: 10.1159/000506735

 

Confocal Raman Microscope 

Purchased: 2012
Latest upgrade: 2014 – TrueSurface module

Responsible person: Dr. Václav Ranc  (vaclav.ranc@upol.cz), since 06.2021

Technology: Olympus LSM microcope with WITec Raman detector module

Special features: 

  • Excitations: 405, 532, 633 and 785 nm.

Established techniques: 

  • Raman mapping, TrueSurface, RayShield.

Selected publications using the technology:

Medvedikova, M., Ranc, V., Vanco, J., Travnicek, Z., Anzenbacher, P., Effect of copper(II) mixed-ligand quinolinonato complexes on the activity of selected cytochromes P450, Journal of biological inorganic chemistry, submitted.
Ranc V., Kalita, O., Pavlacka, O., Vaverka, M., Discrimination of resected glioma tissue using surface enhanced Raman spectroscopy and Au@ZrO2 nanocomposite, Frontiers in chemistry, in preparation. 

 

NanoScan PET/MRI 3T (Mediso) 

Purchased: 2022
Latest upgrade: 

Responsible person: Dr. Miloš Petřík (milos.Petrik@seznam.cz), 

Technology: Hybrid PET/MRI small animal imaging systems by Mediso

Special features: 

  • Full-scale, quantitative PET combined with a robust, cryogen-free MRI (3 Tesla magnet)
  • In-line PET subsystem with dynamic scanning
  • Quantitative imaging of mice, rats and even larger animals. 

Established techniques: in vivo cancer studies for mouse and rat involving:

  • Pharmacokinetics
  • Pharmacodynamics
  • ADME
  • receptor binding
  • radiolabelled tracers
  • infection imaging

Selected publications using the technology:

 

Fluorescence, confocal and super-resolution microscopy

Responsible person: Martin Mistrik Ph.D.

Equipment:
  • Cell Observer - Spinning disk confocal microscope from Zeiss with live cells support
  • LSM780 + Elyra PS.1 + 355nm laser – Confocal microscope by Zeiss together with Elyra PS.1 super-resolution module allowing TIRF, SR-SIM and PALM techniques with live cells support. Also equipped by 355nm UV-A laser for DNA damage induction in live cells
  • ScanR - High throughput screening fluorescent confocal microscope by Olympus


 

 

Raman and infrared microscopy

Responsible person: Dr. Václav Ranc  (vaclav.ranc@upol.cz), since 06.2021

Equipment:
  • Alpha300 R - Confocal Raman Microscope and Raman Imaging System by Witec equipped by three excitation lasers 405nm, 532nm and 785nm
  • Nicolet iN10 FT-IR – Infrared microscope by ThermoScientific, equipped with external module iZ10