FINALS: HISTOPATHOLOGY Flashcards
IMMUNOHISTOCHEMISTRY
- X ___, special procedure in ___
- involves ___
- routine
- disease diagnosis
- antigen-antibody reactions
PURPOSE OF IHC:
- Most common form of Ab used in IHC
- For identification of tissue / cellular antigens / phenotypic markers
- Detection of organisms (microbes: cause diseases) in cytologic procedures (sputum, body fluids, FNA)
ANTIGENS (found in ___)
- cells
- to be detected
ANTIBODIES (class of ___, ___)
Most common Ig used?
- serum proteins
- immunoglobulins
- IgG
TYPES OF ANTIBODIES USED IN IHC
A. ___
- produced by ___
- react w/ ___
- from ___
Main Source:
Other Source:
- POLYCLONAL
- different cells
- various isotopes
- laboratory animal sources
- RABBITS - immunized to produce Abs
- GOATS, PIGS, SHEEP
TYPES OF ANTIBODIES USED IN IHC
B. ___
- more ___
- produced from ___
- reacts only w/ ___
Main Animal Source:
B. MONOCLONAL
- specific
- individual clone of plasma cell
- 1 specific type of isotope
- MICE
LABELLING OF ANTIBODIES
1. USE OF ___
Most widely used?
Alternative?
- ENZYMES
- HRP (HORSE RADISH PEROXIDASE)
- AP (ALKALINE PHOSPHATASE)
Required in use of enzymes to see ___?
Required by chromogens; ___?
- CHROMOGENS -> color developers
- positive result
- SUBSTRATE
- consumed by enzyme
Ab + ___ to react w/ antigen
E + C + S
Enzyme: HRP (HORSE RADISH PEROXIDASE)
Substrate: ___
Chromogen:
1. ___
Color:
Suitable Counterstains:
- ___
Color:
Suitable Counterstains:
Enzyme: HRP (HORSE RADISH PEROXIDASE)
Substrate: HYDROGEN PEROXIDASE (H2O2)
Chromogen:
1. DAB (3,3’ DAB diaminobenzidine)
Color: Brown
Suitable Counterstains: Hematoxylin, Methyl green, Methyl blue
- AEC (3-amino-9-ethylcarbazole)
Color: Brick red
Suitable Counterstains: Hematoxylin, Methyl blue
Enzyme: AP (ALKALINE PHOSPHATASE)
Substrate: ___
Chromogen:
1. ___
Color:
Suitable Counterstains:
- ___
Color:
Suitable Counterstains:
Enzyme: AP (ALKALINE PHOSPHATASE)
Substrate: Phosphate
Chromogen:
1. BCIP (5-Bromo-4-Chloro-3-Indolyl Phosphate) / NBT (Nitro Blue Tetrazolium)
Color: Bluish violet / Black violet
Suitable Counterstains: Nuclear fast red, Brilliant green
- FAST TR RED SALT
Color: Rose color
Suitable Counterstains: NONE
Used to observe end products?
Fluorochrome dye used?
- LIGHT MICROSCOPE
- FLUORESCENCE MICROSCOPE
HEMATOXYLIN AS COUNTERSTAIN
- ___; ___
MAYER’S OR HARRIS HEMATOXYLIN - ___
MAYER’S - ___
HARRIS - ___; ___
- traditional counterstain; nuclear stain
- Immunohistochemistry
- Reddish violet
- Red; commonly used bc tissue morphology are well-defined
INCUBATION - ___
Incubation time & temperature to link Ab w/ ___
- required to link Ag to Ab
- 30-60 MINS AT ROOM TEMPERATURE
- peroxidase
LABELLING OF ANTIBODIES:
2. USE OF ___
- requires ___
Most common:
- FLUROCHROME LABEL
- FLUORESCENCE MICROSCOPE
- FITC (FLUORESCENCE ISOTHIOCYANATE)
LABELLING OF ANTIBODIES:
3. USE OF ___
GOLD - ___ end-product (under ____
- COLLOIDAL METALS
- pink
- light microscope
LABELLING OF ANTIBODIES:
4. ___
To detect ___ (traditionally use ___)
- Can bind w/ ___; can also be ___
Source:
- RADIO LABEL
- LECTINS
- antigen
- antibody
- tissue carbohydrates
- labeled
- PLANT / ANIMALS
SPECIMENS FOR IHC:
1. ___
- for ___
- done ___ (while ___)
- tissue prepared as ___ & must be fixed for a few sections using ___ / ___
Preparation: Cut tissue using ___ / ___
- FROZEN SECTION
- rapid diagnosis
- intraoperatively
- patient is in OR
- cryostat section
- METHANOL / ACETONE
- freezer microtome / CRYOSTAT
PURPOSE OF FIXATION:
- To preserve the position of antigens being detected
- To prevent destruction of labile antigenic sites
SPECIMENS FOR IHC:
2. ___ LIKE ___ & ___
Preferred fixative for IHC?
Required for processed specimens?
Purpose:
- PROCESSED SPECIMENS
- Formalin fixed, Paraffin embedded
- FORMALDEHYDE
- ANTIGEN RETRIEVAL
- Increase accessibility of Ag to Ab
2 GENERAL METHODS OF ANTIGEN RETRIEVAL:
1. ___
- ___ / ___
MOST COMMON ENZYMES USED:
Both can break ___
Forms crosslinks?
- HIER (HEAT INDUCED EPITOPE RETRIEVAL)
- PIER (PROTEASED INDUCED EPITOPE RETRIEVAL) / PROTEOLYTIC ENZYME RETRIEVAL
- TRYPSIN, PROTEASE
- formalin crosslinks
- 10% FORMALIN AS FIXATIVE
PRE-TREATMENT
BLOCKING - only for ___, X ___
1. ___
- ___
- uses ___
- ___
- all ___ & reduces ___
- uses ___
- processed specimen
- frozen section
- PEROXIDASE BLOCK
- endogenous peroxidases
- 3% H2O2 (HYDROGEN PEROXIDE)
- PROTEIN BLOCK
- non-specific sites
- background
- 10% NORMAL SERUM
PURPOSE OF USE OF CONTROL AS REQUIREMENT IN IHC:
- To test for specificity of Abs involved
- To prevent misinterpretation due to false + & false - results
POSITIVE CONTROL - tissue section having the ___
NEGATIVE CONTROL - omit ___ from ___ / replace ___ by an __ directed against an ___
INTERNAL TISSUE CONTROL / BUILT IN CONTROL -contains ___ not only in the ___ but also in ___
- Ag being detected
- primary Ab
- staining sched
- primary Ab
- Ig
- unrelated Ig
- target Ag
- tissue elements
- adjacent normal tissue elements
DETECTION USING LABELED ENZYME ANTIBODY
1. ___
- use of ___
- Add ___
- ___
- ___ of traditional (direct); ___ of direct
- involves addition of ___; more ___ - ___
- ___
- requires use of ___
PRIMARY Ab -
SECONDARY Ab - will attach / bind to ___ -> ___ (better ___)
- DIRECT
- only 1 Ab that is labeled
- labeled Ab to Ag being detected
- EPOS (ENHANCED POLYMER ONE-STEP STAINING METHOD)
- improvement
- type
- DEXTRAN POLYMER
- sensitive
- INDIRECT
- two-step
- 2 Abs
- specific to Ag being detected
- Fc portion of primary Ab
- INCREASING SENSITIVITY
- signal amplification
ENZYMATIC METHOD - ___ are labeled w/ ___ before ___; after reacting w/ ___, ___ is formed
___ - catalyzes ___ to yield a ___ w/c can be analyzed using either ___ / ___
- Abs
- enzyme
- reaction
- targeted Ag
- Ag-Ab COMPLEX
- Ag-Ab COMPLEX
- substrate
- colored product
- LM / EM
ADVANTAGES OF ENZYMATIC METHOD:
- Standard samples can be stored for a long time
- Hematoxylin can be used as a counterstain
- End product can easily be identified using LM / EM
- Will not require fluorescence microscope
AUTOMATION IN IHC:
- Greater consistency of staining
- Faster & more accurate
- Less manpower
- Decreased use of reagent
TUMOR MARKERS - for ___ & ___ of cancers
- found in ___; markers for ___
- detection & staging
- tissues
- malignancies
A. EPITHELIAL TUMOR MARKERS
1. KERATIN - for ___; ___
CARCINOMA - ___ in origin
SARCOMA - __ in origin
Ex:
CK 7 (CYTOKERATIN 7) - ___
CK 20 - ___
X all types of carcinoma will be ___
- EMA (EPITHELIAL MEMBRANE ANTIGEN) - ___
- determine ___; ___ - CEA (CARCINOEMBRYONIC ANTIGEN) - ___
- differentiates ___ from ___
ADENOCARCINOMA - cancer of ___ involving ___
- elevated: ___
- monitor ___ - TTF-I (THYROID TRANSCRIPTION FACTOR I) - differentiates ___ from ___
- found in ___ - PSA (PROSTATE SPECIFIC ANTIGEN) - highly specific for ___
- elevated: ___ - ER/PR (ESTROGEN-PROGESTERONE RECEPTOR) - normally found in ___ but X in ___
- prognosis & treatment: ___
- identification: ___
A. EPITHELIAL TUMOR MARKERS
1. KERATIN - for ___; ___
CARCINOMA - ___ in origin
SARCOMA - __ in origin
Ex:
CK 7 (CYTOKERATIN 7) - ___
CK 20 - ___
X all types of carcinoma will be ___
- EMA (EPITHELIAL MEMBRANE ANTIGEN) - ___
- determine ___; ___ - CEA (CARCINOEMBRYONIC ANTIGEN) - ___
- differentiates ___ from ___
ADENOCARCINOMA - cancer of ___ involving ___
- elevated: ___
- monitor ___ - TTF-I (THYROID TRANSCRIPTION FACTOR I) - differentiates ___ from ___
- found in ___ - PSA (PROSTATE SPECIFIC ANTIGEN) - highly specific for ___
- elevated: ___ - ER/PR (ESTROGEN-PROGESTERONE RECEPTOR) - normally found in ___ but X in ___
- prognosis & treatment: ___
- identification: ___
A. EPITHELIAL TUMOR MARKERS
1. KERATIN - for ___; ___
CARCINOMA - ___ in origin
SARCOMA - __ in origin
Ex:
CK 7 (CYTOKERATIN 7) - ___
CK 20 - ___
X all types of carcinoma will be ___
A. EPITHELIAL TUMOR MARKERS
1. KERATIN - for epithelial tissue cells; CARCINOMA
CARCINOMA - epithelial in origin
SARCOMA - connective tissue in origin
Ex:
CK 7 (CYTOKERATIN 7) - CANCER OF LUNGS, UTERUS, OVARIES
CK 20 - COLON & STOMACH CANCER
X all types of carcinoma will be (+) for CK
A. EPITHELIAL TUMOR MARKERS
2. EMA (EPITHELIAL MEMBRANE ANTIGEN) - ___
- determine ___; ___
A. EPITHELIAL TUMOR MARKERS
2. EMA (EPITHELIAL MEMBRANE ANTIGEN) - ADENOCARCINOMA
- determine site of tumor; BREAST, LUNG, KIDNEY CANCER
A. EPITHELIAL TUMOR MARKERS
3. CEA (CARCINOEMBRYONIC ANTIGEN) - ___
- differentiates ___ from ___
ADENOCARCINOMA - cancer of ___ involving ___
- elevated: ___
- monitor ___
A. EPITHELIAL TUMOR MARKERS
3. CEA (CARCINOEMBRYONIC ANTIGEN) - oncofetal
- differentiates ADENOCARCINOMA from MESOTHELIOMA
ADENOCARCINOMA - cancer of epithelial tissues involving glands
- elevated: CERTAIN CANCERS OF GIT
- monitor COLON, PANCREAS, CERVIX, BREAST CANCER
A. EPITHELIAL TUMOR MARKERS
4. TTF-I (THYROID TRANSCRIPTION FACTOR I) - differentiates ___ from ___
- found in ___
A. EPITHELIAL TUMOR MARKERS
4. TTF-I (THYROID TRANSCRIPTION FACTOR I) - differentiates LUNG ADENOCARCINOMA from MESOTHELIOMA
- found in thyroid neoplasm
A. EPITHELIAL TUMOR MARKERS
5. PSA (PROSTATE SPECIFIC ANTIGEN) - highly specific for ___
- elevated: ___
A. EPITHELIAL TUMOR MARKERS
5. PSA (PROSTATE SPECIFIC ANTIGEN) - highly specific for prostate
- elevated: PROSTATE CANCER, PANCREATIC CANCER, SALIVARY TUMORS; PROSTATE INFECTION
A. EPITHELIAL TUMOR MARKERS
6. ER/PR (ESTROGEN-PROGESTERONE RECEPTOR) - normally found in ___ but X in ___
- prognosis & treatment: ___
- identification: ___
A. EPITHELIAL TUMOR MARKERS
6. ER/PR (ESTROGEN-PROGESTERONE RECEPTOR) - normally found in breast epithelial cells but X in myoepithelial cells
- prognosis & treatment: BREAST CANCER
- identification: METASTATIC BREAST CARCINOMA
B. INTERMEDIATE FILAMENT MARKERS
1. ACTIN - tumors derived from ___
B. INTERMEDIATE FILAMENT MARKERS
1. ACTIN - tumors derived from SMOOTH, CARDIAC, SKELETAL MUSCLES
B. INTERMEDIATE FILAMENT MARKERS
2. VIMENTIN - present in ___ & their counterparts: ___
B. INTERMEDIATE FILAMENT MARKERS
2. VIMENTIN - present in normal mesenchymal cells & their counterparts: SARCOMA, MELANOMA, LEUKEMIA, SEMINOMA SCHWANNOMAS
B. INTERMEDIATE FILAMENT MARKERS
3. DESMIN - highly specific for ___
B. INTERMEDIATE FILAMENT MARKERS
3. DESMIN - highly specific for MYOGENIC TUMORS
B. INTERMEDIATE FILAMENT MARKERS
4. GFAP (GLIAL FIBRILLARY ACIDIC PROTEIN) - confirms diagnosis of ___
B. INTERMEDIATE FILAMENT MARKERS
4. GFAP (GLIAL FIBRILLARY ACIDIC PROTEIN) - confirms diagnosis of ASTROCYTOMA
OTHER MARKERS:
1. HCG (HUMAN CHORIONIC GONADOTROPIN) - ___
OTHER MARKERS:
1. HCG (HUMAN CHORIONIC GONADOTROPIN) - CHORIOCARCINOMA
OTHER MARKERS:
2. AFP (ALPHA FETOPROTEIN) - ___
TERATOMAS - usually ___, aka ___
OTHER MARKERS:
2. AFP (ALPHA FETOPROTEIN) - EMBRYONAL CARCINOMAS & TERATOMAS, HEPATOCELLULAR CARCINOMAS
TERATOMAS - usually benign, aka MONSTROUS TUMORS
OTHER MARKERS:
3. LYMPHOMAS / CD 45 - best screening marker for ___
OTHER MARKERS:
3. LYMPHOMAS / CD 45 - best screening marker for LYMPHOMA
2 MOST COMMON MARKERS TO ASSESS PROLIFERATION OF TUMOR CELLS
- Ki
- MIB-I
