[PDF]     http://dx.doi.org/10.3952/lithjphys.52304

Open access article / Atviros prieigos straipsnis

Lith. J. Phys. 52, 269275 (2012)

A. Dementjeva, J. Kiškisa, and E. Žurauskasb,c
aCenter for Physical Sciences and Technology, A. Goštauto 11, LT– 01108 Vilnius, Lithuania
bFaculty of Medicine, Vilnius University, M. K. Čiurlionio 21, LT– 03101 Vilnius, Lithuania
cPublic Institution National Center of Pathology, P. Baublio 5, LT – 08406 Vilnius, Lithuania
E-mail: andrej@ar.fi.lt

Received 18 November 2011; revised 29 March 2012; accepted 20 September 2012

We demonstrate the capabilities of the coherent anti-Stokes Raman scattering (CARS) microscope and its multimodal operation to image a histological section of human intestinal tissue. The imaging of unstained and stained sections using various nonlinear optical contrasts was performed. The CARS configuration of our microscope allows probing which does not require a preliminary staining of tissue saving the treatment time and providing label-free investigation of original matter. Particular attention was paid to visualisation of unstained tissue. CARS images were recorded in the high wave number Raman spectroscopy region and the spectra of the most distinguished features of images are provided and discussed. Additionally, the two photon excitation fluorescence (TPEF) and second harmonic generation (SHG) contrast mechanisms were used for structural visualisation of both unstained and stained sections of human intestinal tissue. Visualisation of a histological section using all contrast mechanisms mentioned above is analysed and discussed. The research is aimed to draw attention to a potential of CARS/nonlinear microscopy in routine healthcare.
Keywords: coherent anti-Stokes Raman scattering (CARS) microscopy, nonlinear microscopy, medical imaging, histology
PACS: 42.65.Dr, 87.64.M, 78.47.jh

A. Dementjeva, J. Kiškisa, E. Žurauskasb,c
aFizinių ir technologijos mokslų centro Fizikos institutas, Vilnius, Lietuva
bVilniaus universiteto Medicinos fakultetas, Vilnius, Lietuva
cValstybinis patologijos centras, P. Baublio 5, LT – 08406, Vilnius, Lietuva

Darbe pristatome skenuojančio koherentinės antistoksinės Ramano sklaidos (KARS) mikroskopo galimybes vaizdinti histologinius audinių pjūvius. Pateikiame dažytų ir nedažytų žmogaus žarnyno audinių histologinių pjūvių įvairaus netiesinės optikos kontrasto vaizdus. KARS kontrasto vaizdai leidžia atlikti histologinius tyrimus be bandinio dažymo. Ypatingas dėmesys skirtas nedažyto audinio vaizdinimui. KARS vaizdai registruoti didelių Stokso poslinkių srityje. Pateikti ir aptariami ryškiausių struktūrų KARS spektrai. Taip pat pateikti antros harmonikos generacijos bei daugiafotonio žadinimo fluorescencijos vaizdai. Aptarti žmogaus žarnyno audinių dažytų ir nedažytų histologinių pjūvių vaizdai. Tyrimų tikslas – atkreipti dėmesį į KARS mikroskopijos ir netiesinės optinės mikroskopijos potencialą taikant ją įprastos sveikatos priežiūros laboratorinėje praktikoje.

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