Gastrointestinal Lab Manual

Histology
dr. Jan Tambayong, PHK

Gastrointestinal Lab Manual

Introduction Students will work individually. Sit anywhere. This lab manual contains instructions for each one of the GI labs. Each student will be supplied with GI microscopic slides at the beginning of each lab session. Wear a white lab coat every time you enter the lab room. Please return your microslides at the end of every lab. This lab manual is for your personal use only.

Gastrointestinal Lab Manual

Important Information on the Lab Exam The Lab Exam will be a bell ringer exam. There will be an integrated lab exam, called OSPE (Objective Structured Practical Exam) for each block, consisting of disciplines involved in the same block (eg Anatomy, Histology, Pathology, etc), usually consisting of 14 or 15 stations. Each station will have one first-order question and one second-order question.
Second-order questions will include:
Specific features Function Your reasoning

LABORATORY 1
Introduction to the Gastrointestinal System
The purpose of this session is to provide you with an overview of the structure of the upper GI tract, ie. Lips, esophagus and stomach. Identify the similarities and differences between gastric mucosa, small intestinal mucosa and colonic mucosa.

The Lips When examining this microslide, you notice a structure with rounded end, lined by stratified squamous epithelium. One side is covered by skin, with integument of skin like hair follicles, sebaceous glands; this part is the outer part of the lips. Adjacent to this is a curved part: red portion of your lips. Beyond that is the inner part of the lips, entirely composed of mucous epithelia.

Teeth development With small magnification, find the part containing tooth buds, showing the development of teeth. Try to locate :
Enamel/email Dentin Predentin Ameloblast Odontoblast : dark magenta colored : pink colored : slightly pink : closer to enamel : closer to dentin

Sexy Lips

Cleft Lips

Deciduous teeth

The Tongue Notice the unique pattern of the muscles, like the pattern of tikar . The orientation of the muscle fibers are in three directions: horizontal, transverse and vertical, in accordance with its function.

Circumvallate Papillae Observe the surface of the tongue. Youll find a large rounded structure, most of which are embedded beneath the tongue surface. This structure, the circumvallate papillae, is surrounded by a deep groove, lined by stratified epithelia which contains small rounded structures, the taste buds, at its periphery wall. Beneath the groove are serous glands, the Ebners gland, which drain their contents into the groove.

Foliate Papillae Observe the surface of the tongue. Youll see a series of parallel ridges and furrows, the foliate papillae. They are located at the dorsolateral surface of the tongue.

Fungiform Papillae Observe the surface of the tongue. Youll find a mushroom-like structure, the fungiform papillae, bulging above the tongue surface. Next to this structure are situated rows of filiform papillae, which have an elongated, conical structure. Its upper surface contains scattered taste buds.

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Lingual Tonsil At the base of the tongue youll see clusters of solitary lymphnodules. It is covered by stratified squamous epithelium.

Palatine Tonsil Located at the lateral walls of the oropharynx, youll find a pair of lymphoid organs, the palatine tonsils. They are lined with a squamous stratified epithelium, that often becomes so densely infiltrated by lymphocyres that it may be difficult to recognize the real feature of this epithelium.

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The Esophagus This photomicrograph is from the lower third of the esophagus. It is lined with stratified squamous epithelium. The muscularis mucosae is very thick. The muscular layer, the muscularis externa, consists of smooth muscle cells. The muscularis externa of the mid portion of the esophagus consists of a mixture of striated and smooth muscle cells. In the upper part of the esophagus, it is striated muscle.

Esophageal-cardiac junction Observe the epithelial lining. At the esophagealcardiac junction, the epithelium from the esophagus, stratified squamous epithelium, is directly transformed into epithelium of the stomach, simple columnar epithelium.

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Layers of GI tract

Stomach

Fundus Gaster

The stomach, a dilated segment of the digestive tract, is divided into 4 regions: cardia, fundus, corpus, and pylorus. The fundus and corpus are identical in microscopic structure. This photomicrograph is of the fundus gaster. The simple columnar surface epithelium is invaginated to form gastric pits (foveola gastrica). These pits extent to 1/5 of the mucosa thickness into the lamina propria. Cells of the surface epithelium secrete an alkaline mucus, forming a thick gel layer, that protects them from the strong acid. At the base of the gastric pits open several branched tubular gastric (fundic) glands. The neck of the gland consists of stem, mucous neck and parietal (oxyntic) cells. The base of the gland contains parietal, zymogenic (chief) and enteroendocrine cells. Note the large acidophilic cells; these are the parietal cells of the fundus gaster, which secrete HCl and an intrinsic factor for the absorption of vit. B12

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Pilorus gaster

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LABORATORY 2
Identify the histological characteristic appearance of the gastrointestinal tract from the stomach to the recto-anal junction, ie. duodenum, jejunum, ileum, colon, appendix, and recto-anal junction. Identify histological features of small and large intestines.

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Pylorus gaster. The surface epithelium form deep gastric pits, reaching of the mucosa thickness. The glands are short and coiled, consisting of mucus secreting cells. Gastrin (G) cells which release gastrin, are intercalated among the mucous cells. Gastrin stimulates the secretion of acid (HCl) by the parietal cells Enteroendocrine (D ) cells secrete somatostatin, which inhibits the release of other hormones, including gastrin.

The Small Intestine The lining of the small intestine shows a series of permanent folds, plicae circulares (Kerckring), consisting of mucosa and submucosa, having a semilunar, circular or spiral form. Besides that there are outgrowth of the mucosa, forming intestinal villi, consisting of epithelium and lamina propria. This photomicrograph shows many cross-sections of intestinal villi; plica is not seen here. These crosssections consist of absorptive, tall columnar cells , interspersed with mucus-secreting goblet cells , and lamina propria.

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The duodenum In the duodenum, the intestinal villi are leafshaped. Plica Kerckring, if any, is very short. In the submucosa are clusters of coiled, tubular glands, Brunners gland. They may extent into the lamina propria. Their cells are of mucous type; their product is alkaline and acts to protect the duodenal mucous membrane against the effects of acid gastric juice.

Jejunum In this segment of the digestive tract, the plicae circulares Kerckring are the most developed, containing many intestinal villi. At the basal portion of the intestinal glands (also called crypts of Lieberkhn), are exocrine cells with large eosinophilic granules in their apical cytoplasm; these are the Paneth cells. The granules contain lysozyme, with antibacterial activity.

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Ileum This segment of the digestive tract contains intestinal villi, sometimes part of a plica Kerckring. In the lamina propria and submucosa are aggregates of lymphoid nodules known as Peyers patches (plaque Peyer). Each patch contains 10 to 200 nodules and is visible to the naked eye as oval areas on the antimesenteric side of the intestine; they are devoid of villi. There are about 30 patches in the human. Instead of absorptive cells, its covering epithelium consists of M cells.

Colon The colon, or large intestine, consists of a mucous membrane with no folds or villi. The intestinal glands are long and characterized by a great abundance of goblet and absorptive cells, and a small number of enteroendocrine cells. The muscularis layer have fibers of the outer longitudinal layer congregated into 3 thick longitudinal bands called teniae coli. In the anal region the mucous membrane forms a series of longitudinal folds, the rectal columns of Morgagni.

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Appendix The appendix, an evagination of the cecum, is characterized by a relatively small, narrow, and irregular lumen, that is caused by the presence of abundant lymphoid follicles in its wall. It contains fewer and shorter intestinal glands and has no villi or teniae coli.

The recto-anal junction About 2 cm above the anal opening, the intestinal mucosa lining from simple columnar (rectum) is gradually replaced by stratified squamous epithelium (anal eepithelium). In this region the lamina propria contains a plexus of large veins that, excessively dilated and varicose, produce hemorrhoid. The muscular layer is not complete, because only the circular layer persists, its end forming the internal sphincter ani muscle. Surrounding the internal sphincter ani muscle is the external sphincter ani muscle, consisting of clusters of striated muscle fibers.
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LABORATORY 3
Identify the essential histological features of the salivary glands and the liver, with special reference to the organization of parenchyma and stroma. Identify the organization of the bile canaliculi, space of Disse, liver sinusoid and Kupffer cells. Identify the histology and vascular relations of a classic hepatic lobule.
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Parotid Gland It is a branched acinar gland; its excretory portion is composed exclusively of serous cells. As in other large salivary glands, the connective tissue contains many plasma cells and lymphocutes. The plasma cells secrete IgA which forms a complex with a secretory component synthesized by the serous acinar, intercalated duct, and striated duct (intralobular duct) cells. This complex constitutes an immunologic defence mechanism against pathogens in the oral cavity.

Submandibular (submaxillary) Gland It is a branched tubuloacinar gland; its secretory portion contains both mucous and serous cells. The serous cells are the main component of this gland (90%). The presence of extensive lateral and basal membrane infoldings toward the vascular bed increases the ion-transporting surface area 60 times. Because of this, the cell boundaries are indistinct. The cells that form the demilunes secrete the enzyme lysozyme, which main activity is to hydrolyze the walls of certain bacteria.
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Sublingual Gland Like the submandibular gland, this gland is a branched tubuloacinar gland formed of serous and mucous cells. Mucous cells predominate in this gland; serous cells are present exclusively on demilunes of mucous tubules. As in the submandibular gland, cells that form the demilunes in this gland secrete lysozyme.

The duct system of the salivary glands are intercalated ducts (lined by squamous to low cuboidal cells), striated (intralobular) ducts (lined by cuboidal cells with a lot of cytoplasm), and larger interlobular ducts (lined by striated or columnar cells around a large lumen). In this photomicrograph is shown interlobular ducts, with its large lumen, surrounded by connective tissue.

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Liver The liver is the organ in which nutrients absorbed in the digestive tract are processed and stored for use. Most of its blood (70-80%) comes from the portal vein; the smaller percentage is supplied by the hepatic artery. In light microscopic sections, structural units called liver lobules can be seen. In certain animals, eg pigs, the lobules are separated from each other by a layer of connective tissue. This is not the case in humans. In the lobule, the hepatocytes are radially disposed in plates, directed from its periphery to its center. The space between those plates contains capillaries, the liver sinusoids. In addition to the endothelial cells, the sinusoids contain macrophages known as Kupffer cells, on the luminal surface of the endothelial cells. Their main functions are to metabolize aged erythrocytes, digest hemoglobin, secrete proteins related to immunologic processes and destroy bacteria that enter blood through the large intestine.

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LABORATORY 4
Identify the microscopic structure of the pancreatic gland, the exocrine and the endocrine portion of the gland.

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Pancreas It is a mixed exocrine-endocrine gland that produces digestive enzymes and hormones. The enzymes are stored and released by cells of the exocrine portion, arranged in acini. The hormones are synthesized in clusters of endocrine epithelial cells, known as islets of Langerhans. The exocrine portion of the pancreas is similar in structure to the parotid gland (serous cells). The distinction between the two glands can be made based on the absence of striated ducts and the presence of islets of Langerhans in pancreas. Another characteristic detail is that in the pancreas the initial portion of intercalated ducts penetrate the lumen of the acini. Nuclei, surrounded by a pale cytoplasm, belong to the so called centroacinar cells, that constitute the intra-acinar portion of the intercalated duct. Intercalated ducts are tributaries of larger interlobular ducts lined by columnar epithelium. There are no striated ducts in the pancreatic duct system.

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