university of california, san diego histology shared resources

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Tissues and cell types screened during histopathologic analysis: I. Brain: gray and white matter, endothelium, choroid plexus, pituitary, ventricles olfactory lobes, corpus callosum, hippocampus, midbrain, cerebellum, choroid plexus, pia mater II. Heart and circulatory system: cardiomyocytes endothelium and vessels, pericardium of right ventricle, interventricular septum, cardiac muscle, valves, endothelium. III. Lungs: bronchiolar epithelium, vessels and endothelium, stroma, mesothelial layer IV. GastroIntestinal Tract: A. Liver: hepatocytes, Kupffer cells, bile ducts, portal triads and central veins B. Pancreas: acini, Islets (endocrine), ducts, blood vessels and lobules C. Salivary glands, nerves, ganglions, stroma D. Intestines: all areas: mucosal epithelial cells, smooth muscle layers, ganglion cells, blood vessels, a. Esophagus: in addition : squamous and glandular epithelial mucosal layers b. Stomach: in addition :mucosal epithelial cell types c. Duodenum: in addition : Brunners glands d. Ileum: in addition : villi, goblet cells, Peyer’s Patches e. Colon: in addition : goblet cells, Peyer’s Patches V. GenitoUrinaryTract A. Kidney: Cortex/medulla---Glomeruli, tubules— proximal/distal, blood vessels B. Bladder: epithelium, muscle, blood vessels C. Ovaries/Uterus/Fallopian tubes D. Placenta E. Prostate: tubulo-alveolar glands fibromuscular stroma F. Testis, seminiferous tubules, Sertoli cells and germ cells with mature sperm, Leydig cells, vessels, epididymis, VI. Hematopoietic system: A. Spleen: lymphoid nodules, red pulp, macrophages, megakaryocytes, vessels B. Thymus: Cortex, medulla, blood vessels C. Lymph nodes: cortex, medulla, blood vessels, HEVs D. Bone Marrow VII. Endocrine: Adrenal, islets in Pancreas, Parathyroid, Thyroid VIII. Skin and Integument A. Epidermis, dermis with sweat glands, sebaceous glands and hair follicles B. Breast: lobules and ducts C. Bone/ cartilage and fat and skeletal muscle
Tissue Fixation Procedure for Paraffin Processing The following protocol is to be used for tissue sample processed for light microscopy. Tissue fixation 1. The tissue removed from the animal should be placed into marked cassettes and them immediately placed into a container of 10% neutral buffered formalin. 2. Cassettes are to be specially marked with Secureline marker or pencil exclusively as all other marking can be removed from solvents during processing. 3. The volume of fixative must be 15-20 times over the sample volume and the tissue must be completely immersed in the solution. 4. Tissue can remain in fixative at ambient temperature up to 48 hours. No agitation is required 5. Check for clarity of formalin solution day to day. If solution is cloudy, change out the old solution with fresh, especially if tissues will not be processed immediately. 6. The recommended duration of fixation for general mouse tissue that is no larger than 1.5 cm thick is 24- 48 hours. Larger tissue sections should be monitored and fixed for longer periods of time. 7. If glycogen studies are to be made on liver specimens, the fixation time are to be closely monitored or cytoplasmic vacuolation may be conspicuous. 8. Prior to cassetting or prior to processing, tissue samples are to be trimmed for optimal processing and embedding. This size restriction is confined to a maximum height of 3mm and tissue must be in the tissue cassette loosely for ample penetration of paraffin during the processing schedule. 9. Up to one day prior to processing, the tissues can be immersed in 70% alcohol. This step will halt formalin fixation and acclimatize tissue to the first stage of the dehydrating process. However, prolonged exposure to 70% alcohol may have a drying effect on small specimens. 10. Spleen and pancreas should be enclosed between cassette sponges, in cassettes, before fixation to ensure even fixation and sectioning 11. Embryos of differing sizes require specific fixation protocols. Please contact the histology personnel for help prior to harvesting embryos for microscopic examination Recommended Materials: Securline Marker—Fisher Cat. No. 14-905-30 Buffered Formaldefresh—Fisher Cat. No. SF93-4 Freshly prepared 4% paraformaldehyde Bouin’s solution: Sigma Cat. No. H10-1-32 should be used to fix opened embryos and other small slices of organs that need good morphologic analysis. Fix for 6- 8 hours and then transfer to 70% alcohol to “wash out” the yellow before submitting to histology for automated dehydration, paraffin embedding, sectioning and staining. ZincFormaldehyde fixative: to fix organs that will be used for immunohistochemistry stains. Fix thin slices in this fixative obtained from American Master Tech Phone: 209-368-4031. Cat. No. FXZFO
Page 1: Nissi M. Varki M.D., Director. Emai
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Page 11: Date: Immunohistochemistry checks A

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