Biomedical Imaging core facility

TissueFAXSiPlus image cytometry system

The TissueFAXSiPlus inverted imaging systems from Tissue Gnostics is available in our Department (since 2012) for quantitative analysis of histological samples in brightfield (HistoQuest), and of fluorescent live or fixed cell samples (TissueQuest). The automatic control of microscope stage by TissueFAXS Plates/Slides modules allows scanning and reconstitution of entire sample areas for both image analysis and archiving purposes. Samples can be investigated in bright field light (histology staining) using the PixelLink camera or fluorescence using the PCOPixelFly camera. Our services include quantitative cytometry statistical analysis of cell subpopulations based upon specific marker quantitation. The advanced analysis software StrataQuest allows the automatic stratification of analysed tissues sections (i.e. in tumor and stroma regions) based on various cell or nuclei parameters. Dedicated APPs were configured and made available by the TissueGnostics team of software developers in Iasi.

Contact persons: Dr. Livia Sima, Dr. Gabriela Negroiu

The system has benefited of several upgrades that were supported by national grants:

• MiniBioLab: Miniaturized bio-platform for simultaneous melanoma combination therapy and tumorigenicity screening
• Gerontomics: Multi-omics prediction system for prioritization of gerontological interventions 2016-2021
• TG2TARGET:  Role of TG2 in cancer tumor microenvironment for guiding metastasis prevention therapeutic approaches

Technical parameters

HARDWARE

Fully motorized inverted microscopy system for the automated scan and analysis of up to 12 slides or alternatively two cell culture multiwell plates in epifluorescence and brightfield 

Optical Components based on Zeiss AxioObserver.Z1

Inverted optical system – dedicated to cell cultures

Objectives: 

5×, 20×, 40×, 63× Oil

The system is calibrated for each objective, providing fort the calculation of sample area and cell count per mm2. 

Can be equipped with a total of 7 objectives

Filters - 5 filter sets for fluorescence applications:

• DAPI (Ex. 360nm/Em. 462nm)
• Alexa 488/Cy2 (Ex. 488nm/ Em. 520nm)
• Alexa 568/Cy3 (Ex. 568nm/ Em. 603nm)
• Alexa 633/Cy5/Allophycocyanin (APC) (Ex. 633nm/ Em. 651nm)
• Cy5.5 (Ex. 650/45nm; Em. 720/60nm)

Can be equipped with a total of 6 filter sets (simultaneously inserted)

All TissueGnostics fluorescence filters are pixelshiftfree

Ultra-precise motorized stage for 12 slides 

Motorised stage specially adapted to the system, for the automated acquisition and creation of overview images and ROI details. Specific fields of view on the sample can be reacquired.

Equipped for the analysis of: 

• multiwell plates
• paraffin sections
• cryocut sections
• biopsies
• tissue microarrays (TMA)
• cell cultures
• cell smears
• cytospin preparations

Digital high-speed camera for brightfield microscopy (PixelLink), suitable for the acquisition of H&E, IHC, ISH

High-sensitivity digital monochrome (PCO Pixelfly) camera for fluorescence microscopy, suitable for the acquisition of direct and indirect IF, FISH, FRET, FRAP

High performance PC/Workstation for microscope control and analysis - MS Windows 7 

SOFTWARE 

Acquisition module TissueFAXS

• Control software for AxioObserver.Z1
• Microscope stage
• Pixelink color camera
• PCO monochrome camera
• Preview scan (1x to 10x) “Fluorescence”
• Preview scan (1x to 10x) „Brightfield“
• Autofocus for freely selectable magnification and performance in fluorescence mode & brightfield mode
• Generation of stitched overview images with selectable resolution 
• Automatic acquisition of brightfield images and fluorescence images 

Fluorescence analysis module TissueQuest 

Tissue cytometry in immunofluorescence 

• Innovative software for single-cell identification in tissues
• Dynamic data connection “from cells to dotplot” (forward connection) and “from dotplot to cells“ (backward connection)
• Up to 9 analysis parameters available
• Logarithmic and linear data display
• Copy & Paste functionality using Windows clipboard for image- and data export into other applications (e.g. MS PowerPoint)
• Single-site licence 

Brightfield analysis module HistoQuest

Tissue cytometry in immunohistochemistry 

Innovative solutions for single-cell identification in tissues using brightfield microscopy Measurement of characteristic chromogen-mediated cellular staining patterns 

• Results presented in editable scattergrams
• Dynamic data connection “from cells to dotplot” (forward connection) and “from dotplot to cells“ (backward connection)
• Up to 13 analysis parameters available
• Logarithmic and linear data display
• Data evaluation and data processing in dotblots and tables
• Copy & Paste functionality using Windows clipboard for image- and data export into other applications (e.g. MS PowerPoint)
• Single-site licence

Publications

Publications citing our TissueFAXSiPlus system:

• Sima LE, Chen S, Cardenas H, Zhao G, Wang Y, Ivan C, Huang H, Zhang B, Matei D. Loss of host tissue transglutaminase boosts antitumor T cell immunity by altering STAT1/STAT3 phosphorylation in ovarian cancer. Journal for ImmunoTherapy of Cancer, 2021
• AF Bonciu, S Orobeti, LE Sima, M Icriverzi, M Filipescu, A Moldovan, A Popescu, V Dinca, M Dinescu. Pyramidal shaped ceria nano-biointerfaces for studying the early bone cell response. Applied Surface Science (Elsevier), 06 August 2020
• LE Sima, G Chiritoiu, I Negut, V Grumezescu, S Orobeti, CVA Munteanu, F Sima, E Axente. Functionalized graphene oxide thin films for anti-tumor drug delivery to melanoma cells. Frontiers in Chemistry (Frontiers Media S.A.), 25 March 2020
• Raluca Ianchis, Claudia M. Ninciuleanu, Ioana C. Gifu, Elvira Alexandrescu, Raluca Somoghi, Augusta R. Gabor, Silviu Preda, Cristina L. Nistor, Sabina Nitu, Cristian Petcu, Madalina Icriverzi, Paula E. Florian and Anca M. Roseanu. Novel Hydrogel-Advanced Modified Clay Nanocomposites as Possible Vehicles for Drug Delivery and Controlled Release. Nanomaterials (MDPI), 13 December 2017
• M Icriverzi, A Bonciu, L Rusen, LE Sima, S Brajnicov, A Cimpean, RW Evans, V Dinca, A Roseanu. Human Mesenchymal Stem Cell Response to Lactoferrin-based Composite Coatings. Materials (MDPI), 18 October 2019
• Madalina Icriverzi, Laurentiu Rusen, Livia Elena Sima, Antoniu Moldovan, Simona Brajnicov, Anca Bonciu, Natalia Mihailescu, Maria Dinescu, Anisoara Cimpean, Anca Roseanu, Valentina Dinca. In vitro behavior of human mesenchymal stem cells on poly(N-isopropylacrylamide) based biointerfaces obtained by matrix assisted pulsed laser evaporation. Applied Surface Science (Elsevier), 28 February 2018
• A.C.Popescu, P.E.Florian, G.E.Stan, G.Popescu-Pelin, I.Zgura, M.Enculescu, F.N.Oktard, R.Trusca, L.E.Sima, A.Roseanu, L.Duta. Physical-chemical characterization and biological assessment of simple and lithium-doped biological-derived hydroxyapatite thin films for a new generation of metallic implants. Applied Surface Science (Elsevier), 06 January 2018
• Grumezescu, A.M. Holban, L.E. Sima, M.B. Chiritoiu, G.N. Chiritoiu, A.M. Grumezescu, L. Ivan, F. Safciuc, F. Antohe, C. Florica, C.R. Luculescu, M.C. Chifiriuc, G. Socol. Laser deposition of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) − Lysozyme microspheres based coatings with anti-microbial properties. International Journal of Pharmaceutics (Elsevier), 07 February 2017
• Rașoga, L. Sima, M. Chirițoiu, G. Popescu-Pelin, O. Fufă, d, V. Grumezescu, M. Socol, A. Stănculescu, I. Zgură, G. Socol. Biocomposite coatings based on Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/calcium phosphates obtained by MAPLE for bone tissue engineering. Applied Surface Science (Elsevier), 31 January 2017
• Mihaela Uta,Livia E. Sima,Patrik Hoffmann,Valentina Dinca, Norica Branza-Nichita. Development of a DsRed-expressing HepaRG cell line for real-time monitoring of hepatocyte-like cell differentiation by fluorescence imaging, with application in screening of novel geometric microstructured cell growth substrates. Biomedical Microdevices (Springer), 09 January 2017
• D. Scarisoreanu, F. Craciun, V. Ion, R. Birjega, A. Bercea V. Dinc, M. Dinescu, L. E. Sima, M. Icriverzi, A. Roseanu, L. Gruionu, and G. Gruionu. Lead-Free Piezoelectric (Ba,Ca)(Zr,Ti)O3 Thin Films for Biocompatible and Flexible Devices. Applied Materials and Interfaces (ACS Publications), 08 December 2016
• Visan, R. Cristescu, N. Stefan, M. Miroiu, C. Nita, M. Socol, C. Florica, O. Rasoga, I. Zgura, L.E. Sima, M. Chiritoiu, M.C. Chifiriuc, A.M. Holban, I.N. Mihailescu, G. SocoL. Antimicrobial polycaprolactone/ polyethylene glycol embedded lysozyme coatings of Ti implants for osteoblast functional properties in tissue engineering. Applied Surface Science (Elsevier), 19 November 2016 
• Popescu-Pelin, F. Sima, L.E. Sima, C.N. Mihailescu, C. Luculescu, I. Iordache, M. Socol, G. Socol, I.N. Mihailescu. Hydroxyapatite thin films grown by pulsed laser deposition and matrix assisted pulsed laser evaporation: Comparative study. Applied Surface Science (Elsevier), 11 October 2016
• A Honcea, R Iulian, AG Pavel, I Ion, LE Sima. Amniotic Fluid Cells Proliferation in Normal and Down Syndrome Subjects. ARS Medica Tomitana (Sciendo), 11 June 2016
• Popa IL, Milac AL, Sima LE, Alexandru PR, Pastrama F, Munteanu CV, Negroiu Gabriela. Cross-talk between DopaChromeTautomerase and Caveolin-1 is melanoma cell phenotype specific and potentially involved in tumor progression.. Journal of biological chemistry (American Society for Biochemistry and Molecular Biology), 06 April 2016