Histopathology and Haematology Unit

Responsible person: Prof. Aleš Hampl, D.V.M., CSc.

Cell and tissue examination will be a key part of the Preclinical Centre’s activities. The purpose of the histopathology and haematology unit will be to provide highly specialized methodological and scientific support for other research units and laboratories.

It will primarily serve for the dissection of experimental animals and the processing of samples from the Preclinical Centre’s workplaces (breeding facility, organoid unit, tissue culture unit, experimental surgery unit, pharmacological-toxicological unit, etc.) and their analysis using classical and advanced histological methods. The output will be a detailed morphological description of samples ranging from the microscopic anatomy of organs, through the histological structure of tissue structures, to the level of cell morphology and ultrastructure, including the visualization of specific antigens. The unit will be logistically organized to ensure the receipt and rapid processing of samples without damage by autolysis. The basic laboratory methods routinely provided by the unit will include automated preparation of fixed tissue blocks, preparation of tissue sections, and their standardized staining using histochemical and immunohistochemical methods. In connection with the organoid unit, the unit will also enable advanced preparation of three-dimensional samples in hydrogels and their fixation and clearing for 3D immunofluorescence visualization.

For blood cell analysis, the unit will be equipped with a veterinary haematology analyser enabling blood count determination, differential leukocyte count, and other specialized haematological examinations of mice and rats, including analysis of body fluids and coagulation parameters. The unit will also enable morphological examinations of cells in peripheral blood smears, bone marrow or trepanobiopsy samples, or cytocentrifuged samples. In addition to basic methods, specific (immuno)cytochemical methods will also be available. The unit will include space for long-term archiving of blocks and slides.

What services/products will the unit provide?

Support with sample collection and fixation

assistance with experiment design
dissection of experimental animals, sample trimming, digital macroscopic recording (MacroPATH, Milestone Medical)
sample fixation (animal and human tissues, engineered tissues)

Histopathological processing of tissues and organs

processing of native or fixed tissue – dehydration, clarification, embedding in blocks
preparation of sections from blocks or frozen tissue
overview and special histochemical and immunocytochemical staining of sections
cooperation in the preparation of samples for fluorescence microscopy
to a limited extent, archiving of material (blocks, slides – e.g. during an ongoing project)

Scanning slides on a fully automatic scanner (Axioscan 7; Zeiss)

Blood cell analysis in laboratory rodents (Haematology Analyzer URIT Smart V5)

haematological analysis of whole blood (20 µl sufficient)
blood count (haemoglobin, haematocrit, MCV, MCH, MCHC, RDW) and cell abnormalities, if applicable
determination of differential count and, if applicable, cell abnormalities
evaluation of haematopoiesis (detection of mature and immature reticulocytes)
platelet examination (count, size, distribution width, etc.)

Determination of biochemical parameters of laboratory rodent blood (Biochemical Immunoassay Analyzer Skyla Solution)

determination of electrolyte levels, lipid spectrum, and sugars
examination of renal functions (ions, urea, creatinine)
examination of liver functions (albumin, AST, ALT, GGT, ALP, bilirubin, ammonia)
examination of pancreatic enzymes (amylase, lipase)
determination of hormone levels (TSH, TT4)
determination of inflammatory marker levels (CRP)

(please discuss specific details in advance with Prof. Hampl; email: ahampl@med.muni.cz)

Multiplex labeling and imaging of up to 18 selected protein targets in human tissue.

Orion (Rarecyte) imaging platform utilizes simultaneous signal detection from 20 channels in a single run
The service includes an initial consultation, design of a multiplex antibody panel, sample staining, and scanning
Detection of selected protein targets is performed using primary fluorescently labeled antibodies
Detection is also possible using a secondary fluorescently labeled antibody
Compatibility with FFPE and “fresh-frozen” tissue sections
Optional- custom conjugation of IgG antibodies suitable for IHC
Basic image analysis—cell segmentation, percentage representation

(please discuss specific details in advance with Dr. Procházková; email: jirina.prochazkova@sci.muni.cz)