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Connective Tissue Lab

Connective Tissue: The Matrix!

Connective tissues are some of the easiest tissue to identify, so lets begin with them. They differ from other tissues in that connective tissues have cells that are spread far apart as opposed to epithelial tissues whose cells reside in close proximity to one another. The extracellular space of connective tissue is composed of a matrix, containing fibers and ground substance. The fibers are composed of various types of protein; the ground substance is anything found between the fibers. This will all become clearer as we examine the tissues. It is both fitting and proper that we begin with the five types of connective tissue proper. Part I: Connective Tissue Proper

Slide #1-1: Areolar connective tissue. (page 55, figure 1, Gartner & Hiatt) H & E stain. Note the thick fibers (collagen) and thin fibers (elastin) contained within this tissue. Various cells are found including fibroblasts, lymphocytes, macrophages and mast cells. Fibroblasts are elongated cells with short cytoplasmic appendages projecting from two ends. They are the fiber producing cells of areolar connective tissue. Lymphocytes appear as small dark disks or ovals. They are responsible for producing antibodies or destroying infected cells. Macrophages and mast cells have granular cytoplasm and can be difficult to distinguish form one another. Macrophages are fixed in areolar connective tissue and are phagocytotic. By contrast, mast cells produce histamine and help induce the inflammatory response when injury occurs. Depending upon the quality of the stain, mast cells and macrophages may also appear as large lightly stained cells which are difficult to distinguish. Under ideal conditions, mast cells display a nucleus that resembles a small white bubble in the middle of the cell surrounded by a granular cytoplasm.

Slide #1-3: Adipose tissue. (page 57, figure 1, Gartner & Hiatt). Adipose tissue consists of cells engorged with fat. The nuclei and cytoplasm of this tissue is compressed into a corner of the cell giving this tissue a bubble-like appearance. Areolar connective tissue may become adipose tissue.

Slide #3-2: White fibrous connective tissue (tendon) (page 57, figure 3, Gartner & Hiatt) H & E stain. This tissue is better known as dense fibrous connective tissue, the regular form. Note the ribbon-like appearance. Fibroblasts of this tissue are often spindle shaped. The fibers are made of collagen.

Slide #2-10: Human scalp. (page 227 Gartner & Hiatt) H & E stain. Find the thick pinkish region that makes the bulk of this slide just below the epidermis. Hair follicles and glands are found nestled within it. This is the dermis of the skin and it is composed of dense fibrous connective tissue, the irregular form. Note the haphazard arrangement of collagen fibers. Leather is made of this tissue.

Slide #3-3 &:3-4. Yellow fibrous connective tissue (elastic ligaments) Mallorys trichrome stain (page 59, figures 1 & 3, Gartner & Hiatt). This tissue is better known as elastic connective tissue. Note the noodlelike quality to the blue stained collagen fibers. The elastic fibers themselves are straight, thick reddish fibers in Mallorys trichrome. With other stains they may appear dark blue, yellow, or with special elastic stain, they appear black . This slide represents the regular form of elastic connective tissue that is found in certain ligaments and the larynx.

Slide #: Aorta. (H & E and Verhoeff stains) (page 59, figure 3, Gartner & Hiatt) Note the thick black fibers in this tissue. These are composed of the irregular form of elastic connective tissue. This slide also illustrates the effects of elastic stain. Between these dark fibers are thin collagen and reticular fibers.

Slide #1-4: Lymph node (page 55, figure 4, Gartner & Hiatt) (Mallorys trichrome stain). The bulk of the lymph node is composed of reticular tissue. Note the fine fiber projecting from dendritic cells. The round cells that make the bulk of the nuclei that can be seen are lymphocytes. The blue tissue is dense irregular connective tissue; the blue color is the effect of Mallorys trichrome. Note that reticular tissue stains black with silver stain as illustrated in your text book.

Part 2: Cartilage

Slide #3-6: Trachea. H & E stain. Explore the trachea and find a broad blue band of tissue that contains little white circles. This is hyaline cartilage. The circles are lacunae, a term derived form the Latin word for lake or pit. Within some lacunae are misty dark bodies, the chondrocytes which are the cells that form cartilage. While all lacunae house chondrocytes in living tissue, many chondrocytes are lost during slide preparation. The term hyaline means glass, and indeed the matrix of the cartilage is glassy and smooth.

Slide #3-7 & 3-8: Elastic cartilage. H & E and Verhoeff stain. Elastic cartilage is very similar to hyaline cartilage, but it contains dark elastic fibers. Elastic cartilage is found in the ear and larynx.

Slide #3-9: Fibrocartilage (Picro-Sirius-hemotoxylin stain). Fibrocartilage often has a wispy fibrous appearance. The chondrocytes are always linearly arranged. Fibrocartilage also stains differently from hyaline and elastic cartilages. On your slide, the fibrocartilage is strained bright blue; in your text book it is stained yellow. These are unusual colors for cartilage. Fibrocartilage accepts different stains because it is acidophilic (preferring acid dyes) while the other cartilages basophlic (preferring basic dyes). This is due in part to a reduction in hyaluronic acid which is abundant in the other cartilages. Its collagen is similar to that found in dense fibrous connective tissue and bone. Other cartilages have their own form of collagen.

Part 3: Bone

Slide #4-9 Ground bone. (page 79, figure 4 & page 81, figure 1, Gartner & Hiatt) Examine the ground bone slide. Note the rings surrounding a dark circle. These rings are called lamellae (literally layers) which are formed from bony matrix. The matrix is composed of a ground substance which is largely calcium phosphate crystals and collagen fibers. Note the spidery little ovals contained within the lamellae. These are lacunae housing osteocytes (bone cells). Osteocytes cannot be seen because ground bone slides are made from non-lining tissue. The lacunae appear to be black because light refracts away from them when viewed under the light microscope. The central dark circle is the Haversian canal which contains blood vessels and, to a lesser degree, nerves in living tissue. The Haversian canal and the lamellae associated with it are called an osteon. The tiny spidery legs extending from the lacunae are called canaliculi. These tiny canals are their life-line for osteocytes imbedded in the bony matrix and extend to other osteocytes as well as to the Haversian canal itself.