Histology Facility

Tissue preparation

HF has two tissue processors for preparing paraffin blocks of solid tissue samples. Both formalin-fixed and Paxgene-fixed tissues can be prepared.

HF has several rotary and sliding microtomes for cutting paraffin blocks (section thickness adjustable to 2-6um). For cutting frozen sections several Leica and Microm cryomicrotomes are available.

Preparation of cultured cells

Cultured cells in suspension can be prepared as sectionable tissue blocks using the thrombin clotting technique. Alternatively, Sakura Cytocentrifuge can be used to prepare cell slides for IHC and IF staining.

Tissue staining

HF has two Kedee stainers for automated hematoxylin-eosin and other routine stains. Slide coverslipping can be done automatically using DAKO coverslipper.

Immunohistochemistry

HF has two Ventana BenchMark and three LabVision autostainers for automated immunohistochemistry. Antigen retrieval can be done with a PT Module, Retriever 2100, or a microwave oven using ph6 or pH9 buffers.

Immunofluorecence

Immunofluorescence staining can be done using LabVision Autostainer or manually using various staining racks available. Of these, JL-Stainer is well-suited for staining of 1-10 IF slides.

In situ hybridization

For in situ hybridization, HF has a fully automated PathCom stainer, which allows IHC, mRNA in situ and DNA in situ hybridization (both CISH and FISH). PathCom has built-in deparaffinization, antigen retrieval as well as proteolytic digestion. mRNA in situ has been done using the RNAscope protocol and reagents.

Tissue microarrays

Tissue microarray paraffin blocks can be prepared using the 3D Histech TMA Master. The system allows preparation of 0.6-2 mm diameter tissue cores punched and inserted in 1-2 recipient TMA blocks. Sectioning and staining of TMA blocks follows the principles of conventional tissue samples.

Light microscopes

HF has several Olympus BX and Leica DM microscope for slide inspection. Some of these are equipped with digital cameras for snapshot imaging.

Fluorescence microscope

HF has a Zeiss Axioplan microscope for multicolor fluorescence imaging and image capture. The microscope is equipped with ApoTome device, which allows structured illumination principle for generating sharp sections-within-section confocal-type images with all wavelengths. The microscope is operated with AxioVision software.

DNA and RNA extraction from paraffin blocks

HF has a QIAGEN Qiacube device for extraction of DNA or RNA from frozen or paraffin block tissue sections. For paraffin block RNA extraction, PAXgene tissue fixation is recommended.

Laser capture microdissection

Veritas Arcturus Laser Capture system is available for isolation of cell islets or single cells from cell cultures and frozen tissue sections. The dissected material can be studied with molecular biology methods (e.g. PCR-based).

 

Virtual microscopy

Virtual microscopy stands for acquiring and handling digitized versions of microscope glass slides, which are in turn commonly referred to as “virtual slides” or “whole-slide images (WSIs)”. Virtual slides are montage images consisting of up to thousands of high resolution digital micrographs.

See our website http://jvsmicroscope.uta.fi for basic introduction.

Virtual slide scanning systems

HF has two virtual slide scanners, (SlideStrider, Jilab, Finland), which enable scanning of up to 200 microscope slides in one batch. Alternatively, scanning of 2×3 inch macroslides is also possible. Scanning resolutions 0.31 um/pixel (using 10X lens) and 0.16 um/pixel (using 20X lens) can be selected by the user. Scanning with oil immersion lenses is also possible.

Both scanners allow scanning with epifluorescence illumination using blue, green and red emission wavelenghts. The scanning output image format is jpeg2000. The viewing software for virtual slides can be downloaded from our website.

Virtual microscopy scanning system for microwell plates

Microwell plates can be scanned using a scanning system mounted in an inverted microscope (Olympus IX71) or using EVOS FL Auto. These systems allow scanning of cells grown on Petri dishes and 24-96 well microplates. The basic illumination system is phase contrast but fluorescent illumination (with motorized filter changer) is also possible.