Connective CH 8 Flashcards
Connective and Muscle Tissue Stains
- Connective tissue is ¼ of the basic tissues and functions to provide metabolic support for other tissues and organs
- Connective tissue neoplasm malignant tumors are termed “scarcomas”
- Consists of 3 different components: fibers, cells, and amorphous ground substances
- Connective tissue proper
- Cartilage
- Bone
- Blood
Connective tissue proper: Collagen:
- Collagen: provide strength, very eosiniophilic, found in tendons organ capsules, and the dermis.
- Is birefringent upon polarization, demonstrated using Masson and Gomori trichrome, and van Gieson stain.
Connective tissue proper: Elastic fibers:
◦ Elastic fibers: present in most fibrous connective tissue. Have a flexible property allowing tissue to stretch.
- Demonstrated by Verhoeff iron hematoxylin, Weigert
resorcin fuchsin, orecein, or Gomori aldehyde fuchsin stains
Connective tissue proper: Reticular fibers
- Reticular fibers: specific type of collagen fiber. These fibers are ARGYROPHILIC.
- This is, how the fibers have the ability to ADSORB silver from a solution, but require a chemical reducing agent
Cells of connective tissue proper
- Fibroblasts: most common, produce connective tissue fibers.
- Mesenchymal cells: develop into various cell types if needed
- Adipose: fat, store lipid, stain through what?
- Mast cells: contain histamine and heparin,
similar to basophils - Macrophages: “big eater” cells, not normally
demonstrated in histopathology lab - Plasma cells: derived from b lymphocytes
- Blood cells: all types
Basement membrane:
Basal lamina
- Basal lamina, seperates epithelium from underlying connective tissue.
- Provides physical support for epithelium, mainly demonstrated by carbohydrate stains, however silver techniques in this chapter
can also be used to stain for basement membranes
Muscle tissue
- ¼ of the basic tissue type
- Skeletal muscle: striated, and voluntary. Actin and myosin are major contractile
components, PTAH can demonstrate cross striations - Cardiac muscle: striated, but involuntary. Centrally located nuclei, and contain
interalated discs - Smooth muscle: non striated, and involuntary. Centrally located nuclei, with
different ratio of actin and myosin.
Connective and Muscle Tissue
Stains
- 14 total
1.) Masson trichrome
2.) Gomori 1-step trichrome
3.) Van Gieson
4.) Verhoeff
5.) Gomori Aldehyde Fuchsin
6.) Movat Pentachrome
7.) Gomori silver
8.) Gordon and Sweets silver
9.) Methenamine-silver
10.) Mallory PTAH
11.) Oil Red O
12.) Sudan Black
13.) Osmium Tetroxide
14.) Toluidine Blue
Masson Trichrome Stain
- Demonstrates differentiation between collagen & muscle
- Sections cut at 4 to 5 microns
- Control tissue: Uterus, small intestine, appendix or fallopian tube
Masson Trichrome Stain
- Reaction is based on three dyes:
1.) Beibrich scarlet – stains cytoplasm, muscle & collagen
2.) Phosphotungstic acid removes Biebrich from collagen
3.) Aniline blue – stains collagen
4.) Weigert’s Hematoxalyn – stains nuclei
Bouin Fixative used as mordant*
- Results:
- Nuclei: Black
- Cytoplasm, keratin & muscle: Red
- Collagen & mucus: Blue
Gomori One Step Trichrome
- Demonstrates differentiation between collagen & muscle
- Sections cut at 4 to 5 microns
- Control tissue: Uterus, small intestine, appendix or fallopian tube
Gomori One-Step Trichrome
Uses combined ingredients
- Chromotrope 2R
- Fast-green FCF (or aniline blue)
- Weigerts Hematoxylin
Bouin Fixative used as mordant
Results:
- Nuclei: Black
- Cytoplasm, keratin & muscle: Red
- Collagen and mucus: Green (or Blue)
Verhoeff – van Gieson
- Verhoeff Elastic Stain: Demonstrates elastic fibers
- Van Gieson:
- Connective tissue stain demonstrating collagen
& muscle - Primarily used as COUNTERSTAIN to Verhoeff
Verhoeff – van Gieson
- Section tissue at 4 to 5 microns
- Control tissue used:
1.) Verhoeff – Section of aorta embedded on edge
(or cross section of large artery).
2.) van Gieson – Uterus, small intestine, appendix
or fallopian tube
Verhoeff Elastic Stain
- Ingredients used:
- Alcoholic Hematoxylin
- Ferric Chloride
- Lugol Iodine
- Results:
- Elastic fibers – Blue black to Black
van Gieson
- Ingredients used:
Ingredients used:
- Picric acid
- Acid Fuchsin
- Weigert’s Hematoxylin
** Picric Acid must remain acidic to have sharp contrast
between collagen and muscle**
Results:
- Nuclei – Black
- Collagen – Brilliant red
- Muscle & cytoplasm - Yellow
Aldehyde Fuchsin Elastic Stain
- Demonstrates elastic fibers
- Demonstrates elastic fibers
- Avoid chromate fixatives
- Cut at 4-5um
- Aorta or skin
Aldehyde Fuchsin Elastic Stain
Reagents:
◦ Reagents:
- Aldehyde fuchsin solution
- Light green
◦ Results:
- Elastic fiber: deep blue to purple
- Other tissue: green
can also demonstrate ground glass appearance in tissues infected with Hepatitis B
virus
Orcein and Reorcein fuchsin
- Orcein , and reorcein fuchsin can also be used. Not common however.
- Orcein stains elastic fibers brownish
- Reorcein Fuchsin stains elastic fibers violet to brownish tinge
- Orcein and Aldehyde Fuchsin methods have been shown to stain Hepatitis B surface antigens
Movat Pentachrome
- Demonstrate mucin, muscle fibrin, elastic, and collagen
- Demonstrate mucin, muscle fibrin, elastic, and collagen
- Cut at 4 – 5 um
- Control tissue: lung ,skin ,or colon
Movat Pentachrome contents
- Alcian blue
- Alkaline alcohol solution
- Iodine-Iodine solution
- Alcoholic hematoxylin
- Ferric chloride
- Working hematoxylin
- Sodium thiosulfate
- Crocein scarlet acid fuchsin
- Phosphotungstic acid
- Alcoholic safran solution
Movat Pentachrome
- Results
Results:
- Nuclei/elastic fibers – black
- Collagen – yellow
- Ground substance/mucin – blue
- Fibrin – intense red
- Muscle – red
Gomori Reticular Stain
- Demonstration of reticular fibers
- Demonstration of reticular fibers
- Section at 4 to 5 microns
- Control tissue used: Liver
Gomori Reticular Stain
CAN NOT USE METAL FORCEPS!!
- Silver stain technique used
- Silver stain technique used.
- Oxidation: produces aldehydes
- Sensitization: metal impregnation (not silver)
- Silver impregnation: ammoniacal or diamine silver
- Reduction: residual silver ions reduced by formaldehyde
- Toning: gold chloride, turns from brown to black
3Ag + AuCl3 -> Au + 3AgCl - Unreduced silver removed: prevents nonspecific silver
to stay bound - Counterstain: provides contrast
