LABORATORY EXERCISE 1 - Wofford College



LABORATORY EXERCISE 1

EPITHELIAL TISSUE

indexed to Ross &Pawlina Histology: A Text and Atlas (6th edition, 2011)

R & P (page number) Slides numbers from histology slide boxes

plate #, & figures

1. Simple Cuboidal (733)

Plate 76, Fig. 1-2 Slide 2, Kidney

Notes: You need not be able to distinguish the various segments of the nephron (proximal and distal tubules, collecting ducts, etc.) for this lab. We’ll learn the difference in those structures when we investigate the kidney in Lab # 10. For now, we examine the renal tubules as representative of a simple cuboidal epithelium

2. Simple Columnar (661)

Plate 67, Fig. 2 Slide 57, Gall bladder

3. Simple Squamous

Plate 1, Fig. 2. (153) Slide 1, Squamous epithelium (look for cobble-stone appearance)

Note: Most of our slides are lightly stained with silver and are whole mounts which means the tissue has not been sectioned and you’re looking through two layers of simple squamous epithelium above and below a very thin middle layer of connective tissue.

4. Pseudostratified columnar

Plate 2, Fig. 4 (155)

Fig. 5. Slide 4, Trachea with cilia

Slide 72, Epididymis with stereocila

5. Stratified squamous (515)

Plate 42, Fig. 1&3 Slide 5, Tongue

Slide 6, Skin, sole

Slide 73, Vagina

6. Stratified cuboidal & Stratified Squamous

Plate 3, Fig. 2 (157) Slide 7, Scalp

Plate 42, Fig. 1 (515)

Note: The only stratified cuboidal tissue in skin is the duct of sweat glands.

7. Transitional

Plate 3, Fig. 4 (157) Slide 8, Bladder (urinary)

LABORATORY EXERCISE 2

CONNECTIVE TISSUE and ADIPOSE TISSUE

1. Loose Connective tissue (193)

Plate 4, Figs. 1-3 Slide 65, Inactive mammary gland (also note adipose cells)

Slide 73, Vagina

2. Dense Regular Connective Tissue (195) No need to deal with endo-, peri-,

Plate 5, Fig. 1-2 Slide 13, Tendon and epi-tendineum terms.

3. Dense Irregular Connective Tissue (517)

Plate 43, Fig. 3 Slide 6, Skin, sole

4. Elastic fibers (197)

Plate 6, Fig. 2 Slide 10, Areolar tissue spread

Fig. 3. Slide 39, Aorta (H&E does not show elastic lamellae well... look at silver-stained slides if available.)

5. Reticular fibers

Text, p. 171, Fig 6.12 Slide 9, Lymph node Best seen in silver stained slides.

Not visible in H&E stained slides.

Reticular fibers are not easily seen with H&E. Most boxes have slides of lymph nodes stained with H&E. A few boxes have silver-stained lymph nodes that do reveal reticular fibers. Look at the silver-stained lymph nodes from Boxes 1, 7, and 14.

6. Hyaline cartilage (211)

Plate 7, Fig. 1-3 Slide 15, Developing endochondral bone. Ignore bone formation;

Study the lightly basophilic hyaline cartilage.

7. Elastic cartilage (215)

Plate 9, Fig. 1-2 Slide 16, Elastic cartilage

8. Fibrocartilage (217)

Plate 10, Figs. 1 & 2 Slide 17, Fibrous cartilage (our slides don’t look like text figures)

Look for isogenic groups of chondrocytes embedded in dense connective tissue.

Box 9 has a very nice slide with a Mallory stain.

9. Adipose Tissue

Text, p. 258, Fig. 9.2 Slide 6, Skin, sole

Text, p. 505, Fig.15.13B Slide 7, Scalp

LABORATORY EXERCISE 3

BONE and BONE DEVELOPMENT

Because endochondral ossification involved the replacement of hyaline cartilage with bone, it may be useful to review hyaline cartilage (see 6-8 of Laboratory Exercise 2.)

An excellent overview of long bone development is presented Fig. 8.17 p. 236. It is prudent to review this figure as a prelude to understanding endochondral ossification.

1. Compact bone (245)

Plate 11, Figs. 1-3 Slide 18, Bone ground

2. Spongy bone and Compact Bone (247)

Plate 12, Fig. 1-2 Slide 19, Developing intramembranous bone

Correction: Bottom half of upper figure is labeled “C”, should be “CB” for compact bone.

3. Intramembranous ossification (253)

Plate 15, Figs. 1-3 Slide 19, Developing intramembranous bone

Some of our Slide 19s are of developing skull or sternum bones from a rat or mouse. These slides look nothing like Plate 15. If you have such a slide, consult Fig. 8.16 on p. 235.

Other Slide 19s are a coronal section through the head of a fetal mouse or rat. Orient by locating the tongue (a very eosinophilic midline structure, as in Plate 15) and the bilaterally symmetrical nasal cavities with deeply basophilic epithelium separated by a midline plate of hyaline cartilage (not included in Plate 15 Fig.1.) Once oriented, ignore the deeply staining developing teeth in the jaws and the roots of whiskers in the skin which are large ovoid structures.

4. Endochondral ossification (249 and 251)

Plate 13, Figs. 1 & 2 Sl

ide 15, Developing endochondral bone

Plate 14, Figs. 1 & 2 Slide 15, Developing endochondral bone

See also Fig. 8.19 p. 238 for zones of cartilage

LABORATORY EXERCISE 4

Avoid getting immersion oil on microscope lenses other than the 100X objective! It is easy to unintentionally spread immersion oil to the 40X objective which renders that lens useless until it is thoroughly cleaned.

At the beginning of lab, students will be shown the proper technique for using oil immersion.

Each student will examine a blood smear slide and classify 100 leukocytes to gain practice identifying lymphocytes, monocytes, basophils, eosinophils, and neutrophils. Data from all students will be compiled to determine if the percentages of leukocytes counted in our slides agree with percentages stated in the text.

BLOOD

1. Blood

Plate 17, all figures (303) Slide 20, Blood smear (examine with oil @ 100x)

Plate 18, top & middle figures (305) Slide 20, Blood smear

MUSCLE

1. Skeletal muscle

Plate 21, Figs. 1-4 (341) Slide 24, Skeletal muscle (ls) (examine with oil @ 100x)

Plate 22, Figs. 1-2 (343) Slide 25, Skeletal muscle (xs)

2. Cardiac muscle

Plate 24, Figs, 1-4 (347) Slide 26, Cardiac muscle

3. Smooth muscle

Plate 26, Figs. 1-4 (351) Slide 22, Smooth muscle;

Slide 23, Uterus

4. Motor nerve ending

Fig. 11.9 (322) Slide 34, Motor nerve ending

| |Lymphocytes |Monocytes |Neutrophils |Eosinophils |Basophils |

|total | | | | | |

| | | | | | |

| | | | | | |

| | | | | | |

| | | | | | |

|percentage | | | | | |

|observed | | | | | |

|percentage | | | | | |

|expected | | | | | |

|(from text p.302) | | | | | |

LABORATORY EXERCISE 6

BLOOD VESSELS (Consult "Aids for Identification of Blood Vessels" on next page.)

Due to the conflicts arising from the differences in vessel classification between our textbook and the barcode guide to the laserdisk images, we will follow the textbook's criteria for vessel type identification. Use our text/atlas’s classification scheme (summarized on the back of this sheet) and not the naming system on the Downing disc.

Note: The fundic stomach slide is used to search for arterioles, capillaries, and venules. Other types of blood vessels may be present as well. Look in the lamina propria beneath the epithelium for vessels of the microvasculature and even deeper in the layer of dense irregular tissue (called the submucosa) for larger blood vessels.

You can determine whether you’re seeing the vessel in cross section or longitudinal section by examining the nuclear profile of smooth muscle cells. Hint: Think “circular” or “cigar.”

1. Arterioles and venules

Plate 35, Fig. 1-4 (439) Slide 37, Fundic stomach

2. Capillaries

Plate 57, Fig. 2 (613) Slide 37, Fundic stomach

Legend for Figure 2 does not mention capillaries. Look for capillaries which should be numerous in the connective tissue just deep to the gastric glands, and should be just large enough for erythrocytes to squeeze through.

3. Venule

Plate 57, Fig. 2 (613) Slide 37, Fundic stomach

Venules not mentioned in figure legend. Be able to distinguish postcapillary venules from muscular venules based on diameter and number of layers of smooth muscles.

4. Muscular arteries and Veins

Plate 34, Fig. 1-3 (437) Slide 38, Artery and vein

Fig 1 is confusing because it shows a cross-section through two vessels running in parallel that share a common wall. The tunics of the muscular artery are to the left of center and merge with the tunics of the vein right.

5. Elastic artery

Plate 33, Figs. 1-3 (435) Slide 39, Aorta

6. Lymphatic vessels

Plate 35, Fig. 3-4 (439) Slide 51, Ileum (look for lacteals in villus; difficult to see.)

Plate 60, Fig. 2 (619)

AIDS FOR INDENTIFICATION OF BLOOD VESSEL TYPES

Due to the confusion arising from the differences in vessel classification between our textbook and the barcode guide to the laserdisk images, we will follow the textbook's criteria for vessel type identification. Remember: Endothelial nuclei have long axis parallel to vessel lumen.

ELASTIC ARTERIES

massive layer of elastic tissues in the tunica media

internal elastic membrane (IEM) present but may merge with elastic lamellae

MUSCULAR ARTERIES

massive (more than 8) layers of smooth muscle in the tunica media

Internal elastic membrane prominent

External elastic membrane present

SMALL ARTERIES

3-8 layers of circularly arranged smooth muscles in the tunica media

Internal elastic membrane present

wall thickness approximately same as lumen diameter

ARTERIOLES

1-2 layers of circularly arranged smooth muscles in the tunica media

Internal elastic membrane may be present or not

Much smaller lumen than muscular venules

CAPILLARY

wall consists of a simple squamous epithelium (endothelium)

no other tunics present

the nucleus of a pericyte (= histiocyte = Rouget cell) can sometimes be seen.

internal diameter is about 10 micrometers or less.

POSTCAPILLARY VENULES

larger diameter than capillary

wall consists of endothelium plus pericytes with crescent-shaped nucleus

MUSCULAR VENULES

like postcapillary venules but with at most 2 layers of smooth muscle

larger lumen than arteriole

no IEM

VEINS (no attempt to distinguish between large and medium veins)

tunica media much thinner than comparable artery

frequently the lumen is partially collapsed, irregularly shaped and larger than artery

no internal elastic membrane

wall thickness much less than diameter of the lumen

LABORATORY EXERCISE 5

NERVOUS TISSUE

1. Nerve cells of ganglia

Plate 27, Fig. 3 & 4 (391) Slide 29, Spinal ganglion (DRG) (most are silver stained)

We do not have slides of autonomic ganglia.

We will distinguish autonomic ganglia and sensory ganglia during laserdisc review.

Read Figure legends for 1 & 2 to learn about Autonomic Ganglia.

2. Structure of peripheral nerve

Plate 28, Figs. 1-4 (393) Slide 30, Human nerve

Slide 31, Nerve fibers

3. Cerebrum

Plate 29, Fig. 1 (395) Slide 76, Cerebrum (silver stain)

Correction: Substitute “neurons” for “nerve” in text at top of page in yellow box.

4. Cerebellum

Plate 30, Fig. 1-4 (397) Slide 75, Cerebellum

5. Spinal cord

Plate 31, Figs. 1-3 (399) Slide 28, Spinal cord

Slide 27, Nerve cells, ox (a smear prep, not sectioned)

6. Motor nerve ending

Fig. 11.9 (322) Slide 34, Motor nerve ending

7. Neuroglia

see text p. 367-373 Slide 35, Protoplasmic astrocytes

Fig. 12.17 - 12.20 p. 372 Slide 36, Fibrous astrocytes

No slide for microglia

Be sure to identify Schmidt-Lanterman clefts (p. 368 Fig. 12.13), Purkinje cells, pyramidal cells, and molecular layer and granular layer of cerebellum in videodisc images.

Know the differences between silver, Nissl, osmium, and Golgi stains.

Other terms to know: neuropil, white matter, gray matter, dorsal horn, ventral horn, dorsal root, ventral root, ependymal cell, endo- , peri-, and epineurium, neurilema, sheath of Schwann.

LABORATORY EXERCISE 7

LYMPHOID ORGANS and SKIN

1. Lymph node

Plate 37, Figs. 1 & 2 (479) Slide 42, Lymph node

Slide 9, Lymph node, reticular

Plate 38, Figs. 1-3 (481) Slide 42, Lymph node

Slide 9, Lymph node, reticular

2. Spleen

Plate 39, Figs. 1-3 (483) Slide 43, Spleen

Plate 40, Figs. 1-4 (438) Slide 43, Spleen

Note: Unlike the text photographs our spleen slides are not well-perfused which means the small lumens visible in Plate 39 will be full of erythrocytes in our slides.

3. Thymus

Plate 41, Figs. 1-3 (487) Slide 44, Thymus

SKIN

1. Thick skin

Plate 42, Fig. 1 (515) Slide 6, Skin

2. Thin skin

Plate 42, Figs. 2 & 3 (515) Slide 7, Scalp

Plate 43, Fig. 1-3 (517) Slide 7, Scalp

Plate 45, Figs. 1-3 (521) Slide 7, Scalp

3. Sensory nerve endings

Plate 46, Fig. 1-4 (523) Slide 32, Meissner's corpuscle (not obvious on all slides)

Slide 33, Pacinian corpuscle (find with low power)

Lymph nodes

subcapsular sinus trabecular sinus afferent lymphatic vessel

efferent lymphatic vessel lymphatic nodules germinal center

cortex medullary cords medullary sinus

reticular cells post-capillary venules hilus

Spleen

red pulp white pulp central artery

germinal center PALS myofibroblasts

capsule trabecular vein venous sinus

stave cell cords of Billroth

Thymus

epithelioreticular cells cortex medulla Hassall's corpuscles

LABORATORY EXERCISE 8

ORAL CAVITY and DIGESTIVE SYSTEM I (Esophagus, Stomach and Salivary Glands)

ORAL CAVITY

1. Tongue : Review Fig. 16.4 (530) for 4 types of papillae

Plate 49, Figs. 1&2 (559) Slide 5, Tongue

Plate 50, Figs. 1&2 (561) Slide 5, Tongue

2. Teeth

Page 536, Fig. 16.7 Demonstration, Tooth

Page 540, Fig. 16.10 Slide 78, Developing Tooth

Page 541, Fig. 16.11 Demonstration, Tooth

Page 542, Fig. 16.13 Slide 45, Tooth in situ

Page 544, Fig. 16.16 Slide 45, Tooth in situ

Slide 78, Developing tooth

DIGESTIVE SYSTEM

1. Esophagus

Plate 54, Figs. 1 & 2 (607) Slide 46, Esophagus

2. Esophogastric junction

Plate 55, Fig. 1 (609) Slide 47, Esophagus and stomach

3. Stomach: Fig 17.8 (576) for review of gastric mucosa and cell types.

Plate 55, Figs. 2-4 (609) Slide 47, Esophagus and stomach

Plate 56, Figs. 1-3 (611) Slide 37, Fundic stomach

Plate 57, Figs. 1-4 (613) Slide 37, Fundic stomach

Plate 58, Figs. 2&3 (615) Slide 48, Pyloric stomach

Look at tooth slide without microscope: identify gingiva, gingival sulcus, alveolar bone, and alveolus.

LABORATORY EXERCISE 9

DIGESTIVE SYSTEM CONTINUED:

Intestines (large and small), Liver, Pancreas, Gall Bladder

1. Small intestine

Plate 59, Figs. 1&2 (617) Slide 49, Duodenum

Slide 3, Duodenum

Plate 60, Figs. 1-3 (619) Slide 49, Jejunum (Fig 1 is ls, our slides are cs)

Slide 50, Jejunum

Plate 61, Figs. 1-3 (621) Slide 49, Ileum

Slide 51, Ileum

2. Large Intestine

Plate 62, Figs. 1,2,4 (623) Slide 52, Colon

ACCESSORY ORGANS OF THE DIGESTIVE TRACT

1. Liver

Plate 65, Figs. 1&2 (657) Slide 56, Liver Warning: It is difficult to distinguish

Plate 66, Figs. 1&2 (659) Slide 56, Liver hepatocytes in poorly perfused liver tissue.

2. Gallbladder

Plate 67, Figs. 1-4 (661) Slide 57, Gallbladder

3. Pancreas

Plate 68, Figs. 1&2 (663) Slide 55, Pancreas

4. Salivary glands

Plate 51, Fig. 2 (563) Slide 54, Submandibular gland

Plate 52, Fig. 2 (565) Slide 53, Parotid gland

Slide 77, 3 types of salivary glands

LABORATORY EXERCISE 10

RESPIRATORY SYSTEM and URINARY SYSTEM

RESPIRATORY SYSTEM

1. Trachea

Plate 71, Figs. 1-3 (693) Slide 4, Trachea

2. Lung

Plate 72, Figs. 1-3 (695) Slide 58 & 81, Lung

Plate 73, Figs. 1-4 (697) Slide 58 & 81, Lung

Slide 80, Emphysema (note size of alveoli)

Slide 79, Smoker’s lung (look for carbon particles)

URINARY SYSTEM

1. Kidney

Plate 75, Figs. 1-2 (731) Slide 2, Kidney

Plate 76, Figs. 1-4 (733) Slide 2, Kidney

Plate 77, Figs. 1&2 (735) Slide 2, Kidney

2. Ureter

Plate 78, Figs. 1-2 (737) Slide 59, Ureter

3. Urinary bladder

Plate 79, Figs. 1-3 (739) Slide 8, Bladder

LABORATORY EXERCISE 11

MALE REPRODUCTIVE SYSTEM

1. Seminiferous tubules

Plate 86, Figs. 1&2 (819) Slide 41, Testis (oil immersion***)

Page 791, Text figure 22.6

2. Efferent Ductules and Epididymis

Plate 88, Figs. 1&2 (823) Slide 72, Epididymis

3. Ductus (vas) deferens

Plate 89, Figs. 1&2 (825) Slide 60, Ductus deferens

4. Prostate gland

Plate 90, Figs. 1-2 (827) Slide 62, Prostate

5. Seminal vesicles

Plate 91, Figs. 1&2 (829) Slide 61, Seminal vesicle

FEMALE REPRODUCTIVE SYSTEM

1. Ovary

Plate 92, Figs. 1-4 (873) Slide 63, Ovary @@@

Plate 93, Figs. 1-5 (875) Slide 63, Ovary

2. Corpus luteum

Plate 94, Figs. 1-4 (877) Slide 64, Corpus luteum

3. Uterus

Plate 96, Figs. 1&2 (881) Slide 23, Uterus

Plate 97, Figs. 1&2, (883) Slide 23, Uterus

4. Mammary gland

Plate 102, Figs. 1&2 (893) Slide 65, Mammary gland, inactive

Plate 103, Figs. 1&2 (895) Slide 66, Mammary gland, active

*** find spermatogonia, primary spermatocytes, early spermatids, late spermatids, spermatozoa, Sertoli cells, Leydig cells.

@@@ find primordial follicles, primary follicles, secondary follicles, Graafian follicles (may not be present in your slide), and atretic follicles.

Be able to correlate stages in the ovary (follicular and luteal) to stages in the uterus (proliferative, secretory, and menstrual) and the hormones responsible (p. 846.)

LABORATORY EXERCISE 12

ENDOCRINE GLANDS and ORGANS OF SPECIAL SENSE

ENDOCRINE GLANDS

1. Pituitary

Plate 80, Fig. 1-3 (773) Slide 67, Pituitary (hypophysis)

2. Thyroid

Plate 83, Fig. 2 (779) Slide 69, Thyroid gland

3. Adrenal gland

Plate 84, Figs. 1-4 (781) Slide 68, Adrenal gland

The clearest representation of the adrenal gland is in Figure 12.23 on page 768.

ORGANS OF SPECIAL SENSE

1. Eye

Plate 104, Fig. 1 (921) Slide 70, Eye

Plate 105, Figs. 1-2 (923) Slide 70, Eye

Plate 106, Figs. 1-2 (925) Slide 70, Eye

Plate 107, Figs. 1-7 (927) Slide 70, Eye

2. Cochlea

Plate 109, Figs. 1&2 (949) Slide 71, Cochlea

Know the cell types, percentages, and general staining patterns of adenohypophysis as presented in Table 21.2 on p.748.

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