BioMediTech Research Infrastructure

Optical 3D-Imaging Microscopy

The facility offers real 3D imaging of transparent materials and tissues in micrometer resolution. In addition to both bright-field and fluorescence microscopes, the facility has state-of-the-art data processing and analysis workstation available for visualization and 3D-image quantification.

Introduction

Optical 3D-Imaging Microscopy facility offers real 3D-imaging of transparent materials and tissues in micrometer resolution. It is situated at Computational Biophysics and Imaging Group (CBIG) in BioMediTech Arvo 2 building. It offers two different optical 3D-imaging systems: a Selective Plane Illumination Microscope (SPIM) and an Optical Projection Tomography (OPT) microscopes.

The systems are being designed and used for imaging transparent biomaterials such as hydrogels with or without cells but usual applications include visualization of anatomy (phenotyping), gene expression (in situ hybridization), protein distribution (immunohistochemistry or GFP expression), transgenic visualization (LacZ).

Typical specimens include mouse and chicken embryos, mouse/rat tissue and organs, zebrafish, drosophila, plants etc.

In addition to the imaging systems the facility has state-of-the-art data processing and analysis workstation available for visualization and 3D-image quantification.

Equipment

OPT

OPT is the optical equivalent of Micro-CT that produces high-resolution (~5 µm) 3-Dimensional image reconstructions of both fluorescently labeled and non-fluorescent (transmission) specimens ranging in size from 1 mm to 15 mm. In OPT a suspended specimen in an index-matching liquid is rotated through a series of angular positions, and an image is captured at each orientation with a camera. Images collected in each orientation are reconstructed in a similar way as in X-ray CT and the 3D-structure or functionality of the sample is recovered.

The OPT was built in-house by our researchers so it is very adaptable concerning the light sources, filters and objective. Thus, it can be easily modified for imaging different types of samples depending on the area of interest.

  • Light source (bright-field mode):
    Telecentric white LED
  • Light source (fluorescent mode):
    Different LED sources: 365, 470, 530, 590, and 660 nm
  • Detector:
    Optical objectives (5x, 10x)
  • Camera:
    Hamamatsu Orca-Flash 4.0
  • Max. field of view:
    about 3 mm
  • Resolution varies depending on the field of view:
    approximately 5 µm

SPIM

Selective plane illumination microscope (SPIM) is a 3D optical imaging method intended for fluorescent samples. High resolution (<1 µm) combined with selective illumination of the focal plane of the objective makes SPIM a very powerful method for imaging living samples. The illumination path of SPIM is orthogonal to the detection path creating a possibility for imaging very deep into the sample (>1 mm) compared to the confocal microscopes (<200 µm) limited depth capability. This combined with very low phototoxicity caused to the imaged sample makes SPIM a viable option for all long term imaging needs of biological, optically translucent samples. The SPIM system has been designed and built in-house by our researchers. The structure of SPIM microscope is not as flexible to modifications as OPT because the illumination and the detection are strictly tied to each other. Our system has been built with multiple laser lines to provide more options when deciding on the applied fluorescence. Our SPIM system also has an OPT system built-in. The multimodal system is more flexible than the normal SPIM by providing structural transmission images for fluorescence localization. Thus, the system offers unprecedented possibilities in high resolution optical imaging.

  • Light source (bright-field mode):
    Telecentric White LED
  • Light source (fluorescent mode):
    Lasers: 488, 561, and 638 nm
  • Detector:
    Optical objective (20x)
  • Camera:
    Hamamatsu Orca-Flash 4.0
  • Max. field of view:
    about 0,7 mm
  • Resolution:
    approximately 1 µm

Prices

Prices will be defined based on customer needs. Please inquire!

Contacts

Facility Director:
Professor Jari Hyttinen
jari.hyttinen(at)tut.fi
Tel: +358 40 849 0020

Publications

Koskela O, Pursiainen S, Belay B, Montonen T, Figueiras E, Hyttinen J. Computational model for multifocal imaging in optical projection tomography and numerical analysis of all-in-focus fusion in tomographic image reconstruction. EMBEC & NBC 2017, Joint Conference of the European Medical and Biological Engineering Conference (EMBEC) and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics (NBC), Tampere, Finland, June 2017.

Koivisto JT, Koskela O, Montonen T, Parraga JE, Joki T, Ylä-Outinen L, Narkilahti S, Figueiras E, Hyttinen J, Kellomäki M. Texture-property relations of bioamine crosslinked gellan gum hydrogels. EMBEC & NBC 2017, Joint Conference of the European Medical and Biological Engineering Conference (EMBEC) and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics (NBC), Tampere, Finland, June 2017.

Belay B, Koivisto JT, Vuornos K, Montonen T, Koskela O, Lehti-Polojärvi M, Miettinen S, Kellomäki M, Figueiras E, Hyttinen J. Optical Projection Tomography Imaging of Single Cells in 3D Gellan Gum Hydrogel. EMBEC & NBC 2017, Joint Conference of the European Medical and Biological Engineering Conference (EMBEC) and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics (NBC), Tampere, Finland, June 2017.

Soto AM, Koivisto JT, Parraga JE, Silva-Correia J, Oliveira JM, Reis RL, Kellomaki M, Hyttinen J, Figueiras E. Optical Projection Tomography Technique for Image Texture and Mass Transport Studies in Hydrogels Based on Gellan Gum. Langmuir, 32 ( 20 ), 5173 – 82, 2016.

Figueiras E, Soto AM, Jesus D, Lehti M, Koivisto J, Parraga JE, Silva-Correia J, Oliveira JM, Reis RL, Kellomäki M, Hyttinen J. Optical Projection Tomography as a tool for 3 D imaging of hydrogels. Biomedical Optics Express, Vol. 5, Issue 10, pp. 3443-3449, 2014.

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