02 Staining & Immunohistochemistry Flashcards
processes that generally employ a dye or chemical that has an affinity for the particular tissue component that is to be demonstrated
special staining
color of neuropathological:
Vogt’s method for Nissl subtance
purple
color of neuropathological stain:
Gallocyanin (Einarson’s) method for Nissl substance
blue
color of neuropathological stain:
modified aldehyde-thionine pas for nissl substance
blue
color of neuropathological stain:
Bielcschowsky method for neurofibrils
black
color of neuropathological stain:
Bodian’s method for nerve fibers & nerve endings
black
color of neuropathological stain:
Holmes method for nerve cells & fibers
black
color of neuropathological stain:
Luxol fast for myelin
blue
application of immunologic principles and techniques to the study of cells and tissues
immunohistochemistry
most commonly used methods for immunohistochemistry
peroxidase-antiperoxidase immune complex
biotin avidin immunoenzymatic technique
identify the effect on results (false neg/false pos):
antibody is inappropriate, denatured, or used at a wrong concentration
false neg
identify the effect on results (false neg/false pos):
loss of antigen through autolysis
false neg
identify the effect on results (false neg/false pos):
loss of antigen through autolysis
false neg
identify the effect on results (false neg/false pos):
presence of antigen is at a density below the level of detection
false neg
identify the effect on results (false neg/false pos):
cross-reactivity of the antibody with antigens different from the one being sought
false pos
identify the effect on results (false neg/false pos):
nonspecific binding of the antibody to the tissues
false pos
identify the effect on results (false neg/false pos):
presence of endogenous peroxidase in some cellular elements
false pos
identify the effect on results (false neg/false pos):
entrapment of normal tissues by the tumor cells
false pos
four main stages of immunohistochemistry
fixation
antigen retrieval
blocking
antibody labeling and visualization
stage in immunohistochemistry:
to keep everything in its place
fixation
stage in immunohistochemistry:
to minimize pesky background signals
blocking
stage in immunohistochemistry:
to get pretty pictures
antibody labeling and visualization
in conventional immunohistochemistry, single immunohistochemical methods are used through ___
3,3’-diaminobenzidine (DAB)
a horseradish peroxidase (HRP) substrate that is used for secondary HRP-conjugated antibody detection
3,3’-diaminobenzidine (DAB)
the two commonly used chromogens
DBA (brown)
AP (red)
chromogen used for most applications as it provides strong and permanent stains
DAB
chromogen used mainly for skin sections
AP Red
ubiquitous contractile protein responsible for cell motility
actin
oncofetal antigen that is the major plasma component of the fetus
alpha fetoprotein
major sources of alpha fetoprotein
live
visceral endoderm of yolk sac
monoclonal antibody that reacts with high molecular weight glycoprotein
B72.3
differentiates mesothelioma and pulmonary adenocarcinom
B72.3
monoclonal antibody directed against an epitope on the protein molecule of two glycoproteins on the surface of glandular epithelial cells of endodermal derivation
Ber-EP4
differentiates adenocarcinoma and mesothelioma
Ber-EP4
vitamin K dependent protein that constitutes approximately 20% of noncollagenous protein in bone
Bone GLA protein
detected in osteosarcoma and chondrosarcoma
Bone GLA protein
carbohydrate antigen
CA19-9
used in the diagnosis and monitoring of pancreatic carcinoma
CA19-9
positive reaction are obtained in most pancreatobiliary adenocarcinomas and transitional cell carcinoma
CA19-9
transmembrane glycoproteins located in desmosomes
E-Cadherins
in E-cadherins, the ductal carcinoma of the breast will show ___ (posi/neg)
positive
in E-cadherins, the lobular carcinoma of the breast will show ___ (posi/neg)
negative
cytoskeleton associated actin binding protein present in contractile cells
calponin
document the presence of myoid differentiation
calponin
calcium binding protein expressed by various types of mesothelial, epithelial, and stromal cells
calretinin
calretinin differentiates from mesothelioma from ___ ___
lung adenocarcinoma
found in synovial sarcomas, sex cord stroma, adamantinoma, and cardiac myxoma
calretinin
glycoprotein normally detected in the glycocalyx of fetal cells, particularly those of mucin secreting nature
Carcinoembryonic Antigen Antibody
Carcinoembryonic Antigen Antibody is expressed in ___ (small/large) quantities in carcinomas
large
identifies neprilysin
CD10
reactive for a variety of female genital tract tumors
CD10
positive in renal cell carcinoma, and solid pseudopapillary tumor of the pancrease
CD10
recognizes hapten x
CD15
differentiates lung adenocarcinoma from mesothelioma
CD15
identifies hematopoietic progenitor antigen ER-MP12 which is identical to PECAM1
CD31
marker for a subset of normal lymphocytes
CD57
reacts with small variety of neural, neuroendocrine and prostatic adenocarcinomas
CD57
mainly used for the diagnosis of Ewing’s sarcoma/PNET
CD99
transmembrane tyrosine kinase expressed by cajal’s interstitial cells, melanocytes, mast cells, and germ cells
c-KIT
identifies c-KIT
CD117
encodes a transcription factor that plays an important role in proliferation and differentiation of intestinal epithelial cells
CDX2
acidic glycoproteins located in the soluble fraction of neurosecretory granules
chromogranin/ synaptophysin
in ___, nearly all types of neuroendocrine tumors are reactive
chromogranin/ synaptophysin
muscle type intermediate filament found in smooth and striated muscle cells
desmin
used in identification of smooth muscle and skeletal muscle tumors
desmin
desmin is abundant in ___ ___ ___
parenchymal smooth muschle
indicator of probable response to EDFR inhibitors
epidermal growth factor receptor
tyrosine kinase anchored on the cell membrane
epidermal growth factor receptor
glycoprotein present in human milk fat globule membrane
epithelial membrane antigen
excellent marker for most normal and neoplastic epitherlium
epithelial membrane antigen
expressed by mesotheliomas, meningiomas, and some mesenchymal neoplasms
epithelial membrane antigen
synthesized in endothelial cells of blood vessels
factor VIII related antigen
used as a marker for endothelial cell differentiation
factor VII related antigen
one of the give major cytoplasmic intermediate filaments
glial fibrillary acidic protein
present in normal, reactive, and neoplastic astrocytes, ependymal cells, and oligodendrocytes
glial fibrillary acidic protein
T/F: glial fibrillary acidic protein is also documented in peripheral nerve sheath tumors
t
acts as an oncogene when over expressed; a membrane protein closely related to EGFR
HER2/ neu
give the expression percentage of HER2:
breast carcinoma
22%
give the expression percentage of HER2:
lung adenocarcinoma
28%
give the expression percentage of HER2:
colorectal carcinoma
17%
give the expression percentage of HER2:
lung squamous cell carcinoma
11%
give the expression percentage of HER2:
gastric adenocarcinoma
11%
T/F: HMB-45 is useful for predicting response to herceptin
F! (it’s HER2/ neu :))
monoclonal antibody obtained from malignant melanoma
HMB-45
identifies oncofetal conjugate associated with immature melanosomes
HMB-45
expressed by neural crest derivative tumors, angiomyolipomas of kidney, tuberous sclerosis
HMB-45
monoclonal antibodies for estrogen, progesterone, and androgen
hormone receptors
normally secreted by syncytiotrophoblast
hCG
two chains of hCG
alpha and beta
used to detect trophopblastic differentiation in germ cells and ectopic hormons
hCG
peptide hormone produced by granulosa cels
inhibin
reacts positively with sex cord stromal tumors of the avary and testes, and steroid producing tumors of the adrenal gland and gonads
inhibin
T/F: keratin is a water soluble intercellular fibrous proteins
F (it is INsoluble and INTRAcellular)
an excellent marker for epithelia differentiation regardless of tumor origin
keratin
keratin:
pancreas, bileducts, and urothelium, 1/3 of gastric carcinoma
CK7+ / CK20+
keratin:
lungs, breast, endometrium, ovary, thyroid, salivary glands, and mesothelioma
CK7+ / CK20-
keratin:
large bowel, merkel cell, 1/3 of gastric carcinoma
CK7- / CK20+
keratin:
adrenal cortex, liver, kidney, adrenal gland
CK7- / CK20-
T/F: Melan A is closely associated with melanosomes
F (not related)
top choice for identification of skeletal muscle differentiation
myogenin
myogenin is strongly expressed in ___
rhabdomyosarcoma
glycolytic dimeric enzymes with isoenzymes found in neurons and neuroendocrine cells, including carcinoids and malignant melanoma
neuron specific enolase
prostate specific antigen has ___ (higher/lower) specificity than prostatic acid phosphate
higher
isoenzyme of acid phosphatase secreted by prostatic epithelium
prostatic acid phosphatase
acidic dimeric calcium binding proteins first isolated in the brain
S100
T/F: S100 doesn’t stain with cytoplasmic staining
F (stains in both nuclear and cytoplasmic staining)
major glycosylates transmembrane protein isolated from neuronal presynaptic vesicles
synaptophysin
expressed in normal, reactive, neoplastic cells of neuroectodermal and neuroendocrine types
synaptophysin
major cytoplasmic intermediate filament that is characteristic of cells of mesenchymal nature
vimentin
T/F: vimentin is not exclusive to cells of mesodermal origin but also in tumors of epithelial or neural in nature
T