Histopathology Flashcards
Derivative of methanol.
Fixes fats, mucin, glycogen
Formaldehyde
-Most widely used for Electron Microscopy
-Fixative of choice for enzyme histochemistry
-2.5% for small tissues
-4.0% for large tissues
Glutaraldehyde
-Fixative for acid mucopolysaccharide
-4% concentration
-For connective tissue mucin
Lead Fixative
-Fixative for mucopolysaccharide, nuclear proteins
Newcomer’s fluid
Best fixation temperature and time for Newcomer’s fluid
3 degrees C for 12-18 hours
Made up of mercuric chloride stock solution and glacial acetic acid before use.
Used for trichrome staining.
Zenker’s fluid
Fixes connective tissue, liver, spleen, nuclei
Zenker’s fluid
Most rapid fixative
Carnoy’s fluid
Fixative for chromosome, lymph glands, urgent biopsies, Nissl granules
Carnoy’s fluid
Most common fixative
Used for tissue photography
5-7% saturated aqueous solution
Renal tissue, connective tissue, muscle, fibrin
Mercuric chloride
Microanatomical and cytological fixative
Used for the identification of cytoplasmic granules
For pituitary gland, bone marrow, blood-containing organs (spleen, liver)
Helly’s fluid or Zenker’s formol
Fixative for hemosiderin (iron pigments:hgb)
Recommended for mitochondria, mitotic figures, chromatin, colloid-containing organs, RBC, Golgi bodies
Regaud’s fluid
Fixative for embryo
Contains picric acid
Glycogen-containing tissue samples
Bouin’s fluid
Fixative for early degenerative processes and tissue necrosis
For Rickettsiae spp identification
Orth’s fluid
Cytological fixative
Tumor biopsies of the skin
Heidenhain’s Susa Fluid
Fixative used for post-mortem changes, research tissues, surgical storage tissues, elastic fibers, iron pigments/deposits
10% Neutral buffered formalin
Fixative for general post-mortem changes
CNS tissues
10% Formol saline
Fixative for routine post-mortem changes
Lipids
Formol corrosive/ Formol sublimate
Better and less messy than Bouin’s
Fixes glycogen
Brasil’s alcoholic picroformol solution
Fixes nuclear structures
Flemming’s fluid with glacial acetic acid
Consists of chromic acid and osmic acid
Fixes cytoplasmic structures
Flemming’s fluid without glacial acetic acid
Golden yellowish brown solution
Fixes glycogen and stains connective tissue
Primary component of Bouin’s
Causes precipitation of all types of proteins
Picric acid
“Conjugated fixative”
Solidifies at 17 degrees Celsius
Precipitates chromosome and chromatin
Glacial acetic acid
Fixative and dehydrant
Used for brain damage (rabies diagnosis)
Cold temperature fixative: -5 to 4 deg C
Demonstrates activities of enzymes (lipases, phosphatases)
Acetone
Fixative and dehydrant
Small hone fragments
Precipitates proteins
Alcohol
Fixes bacteriologic smears
37 to 56 deg c
Enhances fixation
Heat
Histological fixative and decalcifier
1-2% concentration
Fixative for carbohydrates
Strong oxidizing agent
Interferes with hematoxylin dye reactions
Chromic acid
3% concentration
Fixes mitochondria and lipids
Potassium dichromate
6% at 20 deg C
Pale yellow powder
Fixative and stain
For fat and fat derivative sources, adrenal glands, myelin, peripheral glands, cytoplasmic structures
Osmic acid/ Osmium tetroxide
Substitute for glutaraldehyde
Used in electron microscopy and ultrathin microtomy
Strong oxidizing agent
Inhibits hematoxylin dye reactions
Osmic acid / Osmium tetroxide
Fixative for sputum samples
Gemdre’s fluid
Tissue softeners
- Perenyi’s fluid
- Lendrum’s fluid
- Molliflex
- 2% HCl
- 1% HCl in 70% ethanol
Decalcifier
4% aqueous phenol
For fingernails and uterine currettimgs
Lendrum’s fluid
Fixative and a decalcifier
Minute bone spicules
Strong oxidizing agent
Interferes with hematoxylin dye reactions
Chromic acid
Very slow decalcifier
Minute pieces of bones
Sulfurous acid
Very slow, weak decalcifier
Good nuclear staining
Does not require washing out method
Trichloroacetic acid
Decalcifier for autopsy materials
3-24 hours
Formic acid + sodium citrate solution
Fixative and decalcifier
Derivative of formaldehyde
For routine post-mortem research tissues
Moderately in action
Small: 2-7 days
Large: greater than 7 days
Formic acid
Nitric acid-based decalcifier
Silver impregnation of nerve fibers
Consists of chloral hydrates (dissolves in alcohol, H2O + HNO3)
De Castro’s fluid
Most rapid decalcifier: 12-24 hours
Urgent works
Poor nuclear staining
Phloroglucin nitric acid
Excellent nuclear and cytoplasmic staining
PH 4.5
Less distortion of tissue cells
Too slow for routine
Citric acid citrate buffer solution
Most common decalcifier
Routine purposes: Rapid (5-10% concentration)
Fastest
Urgent biopsies
Nitric acid
Decalcifier and tissue softener
Nitric acid-based solution (0.5% chromic acid; 1% HNO3; 100% alcohol)
Slow for bone tissue
Maceration is avoided
Perenyi’s fluid
Tissue softener that turns tissue soapy and swollen
Molliflex
1-5%
Tissue softener and decalcifier
1 & 2%: tissue softener
Recommended for surface tissue block
Inferior than nitric acid: better nuclear staining
Hydrochloric acid
Decalcifier for pieces of bones and teeth
Moderately in action
Cytological studies (nuclear, cytoplasm)
Von Ebner’s fluid
Dehydrant and clearing agent
Causes less shrinkage
6 months exposure: conjunctival irritation -> blindness
Doesn’t dissolve aniline dye
Tetrahydrofuran
Readily miscible in different solvents
Substitute for methanol
Causes minimum shrinkage
Sectioning and smears
Triethyl phosphate
Chemical name of Dioxane
Diethylene dioxide
Dehydrant and clearing agent
Expensive and toxic (not recommended for routine)
Dioxane
Chemical name of cellosolve
Ethylene glycol monoethyl ether (EGME)
Dehydrant preferred for propylene based glycol ethers
Combustible: 110-120 deg F
Does not cause hardening and distortion even if stored for several months
Water-soluble wax
Cellosolve
Fixative and dehydrant
Suitable for urgent biopsies
Harden tissues than ethanol does
Flammable and volatile
Acetone
Miscible with water
Expensive
Does not harden tissues
Isopropanol
Plant and animal tissue microtechniques
Butanol
Fixative and dehydrant
Blood and tissue films and smears
Toxic: breast cancer
Methanol
Most common, cheap, not poisonous
70% : 1 change
95% : 1 change
100% ; 3 changes
Ethanol
Clearing agent most common for routine use
Clearing in embedding and mounting
<3 mm: 15-30 mins
>5 mm: 30-60 mins
Miscible with Canada Balsam
Suitable for urgent biopsies
Unsuitable for brain and lymph nodes
Xylene/Xylol
Lymph nodes, embryo, and nervous tissues (clearing agent)
Toxic into the liver
Chloroform
Clearing agent for CNS tissues and cytological studies (skin, smooth muscle)
Both paraffin and celloidin embedded sections
Disadvantage: slow (5-6 days)
Cedarwood oil
Clearing agent for routine Post-mortem research tissues
Substitute for xylene/benzene
Not carcinogenic but emits toxic fumes
Toluene or Toluol
Clearing agent recommended for urgent biopsies
Carcinogenic: bone marrow damage
Aplastic anemia
Benzene
Ratio of phenol to xylene in carbol-xylene
1:3
Substitute for chloroform
Expensive
Carbon tetrachloride