Stains F-Z
(routine stains, immunostains and molecular markers)
Last revised 25 February 2008
Copyright © 2002-2008 PathologyOutlines.com, Inc.
See also CD Marker chapters
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Note: stains/proteins are in alphabetical order, with numbers before letters, and ignoring dashes and spaces
Go to Stains A-E, including general information
F G H I J K L M N O P Q R S T U V W X Y Z
Interpretation: cytoplasmic stain; endothelium acts as a positive internal control
Positive staining (normal): endothelial cells, megakaryocytes, platelets, mast cells
Positive staining (disease): vascular tumors
Micro images: capillary hemangioma of endometrium (fig 2B); cribriform DCIS of breast - fig 2a: Factor VIII staining shows only rare vascular proliferation; 2b: low MIB1 staining; inflammatory breast carcinoma with factor VIII related antigen staining of dermal lymphatics; factor VIII+ endothelial cells in angiosarcoma of breast
Factor XIIIa
Fibrohistiocytic marker
Micro images: fibrohistiocytic proliferation with dermal nevus (fig c); fibrohistiocytic proliferation with melanoma (fig c)
FADD
Fas (CD95) Associated protein with Death Domain
Not the same protein as Fas
Part of Fas and TNFR1 pathways:
In Fas pathway, death domain portion of Fas recruits FADD
In TNFR1 pathway, TNFR1 binds TRADD, which acts as an adaptor protein to recruit FADD
FAS-FADD and TNFR1-TRADD-FADD recruit FLICE to receptor signaling complex, eventually induce apoptosis
FAK
See focal adhesion kinase
Fas
See CD95
Fascin
55 kDa protein that forms tight and stable cytoplasmic bundles with filamentous actin
Fascin-1: most common type; present in specialized cells with extensive surfaces or migratory potential, such as neurons, glia, dendritic cells, macrophages, skeletal and smooth muscle, endothelial cells; not normal epithelial cells
Fascin-2: in retina; fascin-3: in testis
Actin-bundling protein with important role in cell motility and adhesion
Overexpression in tumors often associated with aggressive disease
Positive staining (disease): carcinoma of biliary tract, breast, colon, lung, ovary, pancreas, skin; follicular dendritic cell tumors, Hodgkin’s lymphoma-classic subtype (highly sensitive), interdigitating dendritic cell tumors Langerhans cell histiocytosis, urothelial carcinoma (noninvasive papillary or invasive)
Negative staining: normal epithelial cells, normal urothelium, benign urothelial lesions
Micro images: synovial sarcoma (positive)
References: Hum Path 2005;36:741
Fas ligand
See CD178
Fat stains
See Oil Red O
Fatty acid synthase
Multifunctional enzyme complex at 17q25 involved in de novo synthesis of saturated fatty acids
Catalyzes conversion of acetyl-CoA and malonyl-CoA into long chain fatty acids via NADPH
Inhibitors are being evaluated as potential therapeutic agents due to toxicity to human cancer cells in vitro
Stronger expression in melanoma vs. nevi; higher for melanoma Clark levels IV/V vs. I/II and Breslow thickness 0.75 mm+ vs. < 0.75 mm; also high in congenital melanocytic nevi (Mod Path 2005;18:1107)
Positive staining (disease): carcinoma of breast, colon, endometrium, ovary, prostate; melanoma
Micro images: various cutaneous melanocytic neoplasms
Fc gamma RIIb
Part of t(1;22)(q22;q11) with lambda light chain
Associated with follicular lymphoma
References: more information #1; #2
Ferritin
Micro images: highly malignant adult hepatic blastoma - fig A: ferritin; B: alpha-fetoprotein; C: HepPar1; D: CK8; E: p53; F: beta-catenin
fes
Tyrosine kinase / signal transducer at 15q25-26
FEV
Gene at 2q33 mutated in Ewing’s sarcoma/PNET
fgr
Tyrosine kinase / signal transducer at 1p36.1-36.2
FHIT
Fragile HIstidine Triad gene
Putative tumor suppressor gene
Deleted in tumors of GI, lung, head/neck
FKHR
Fused with PAX7 gene via t(1;13)(p36;q14) in alveolar rhabdomyosarcoma.
FLI-1
Protein is member of ETS family of DNA binding transcription factors; gene is fli-1, present on #11q24
Involved in cellular proliferation, tumorigenesis, embryologic development of blood vessels
90% of Ewing’s sarcoma/primitive neuroectodermal (PNET) tumors have t(11;22)(q24;q12) which results in fusion of EWS to FLI-1
Sensitive and specific for Ewing’s sarcoma/PNET; also sensitive/specific for vascular tumors vs. sarcomas, carcinomas or melanomas (AJSP 2001;25:1061)
Interpretation: call positive if nuclear staining of 10% of tumor cells (usually is >50%) and positive internal controls of endothelial cells and small lymphocytes (AJSP 2001;25:1061)
Note: other vascular tumor markers (CD31, CD34, von Willebrand factor) are membranous or cytoplasmic stains
Note: cytoplasmic staining present in breast epithelium (benign/malignant) and cutaneous eccrine glands
Uses: differentiate Ewing’s sarcoma/PNET of kidney (positive) from blastema predominant Wilms’ tumor (negative); diagnosis of vascular tumors
Positive staining (normal): endothelial cells, T cells, small lymphocytes
Positive staining (disease): Ewing’s sarcoma/PNET, vascular tumors, lymphomas
Negative staining: blastema predominant Wilms’ tumor; carcinomas, melanomas, non-vascular sarcomas; muscle, nerve, fibroblasts
FLICE/MACH
Member of ICE protease family
MACH = Mort1/FADD Associated CED-3 Homolog
Dominant-negative isoforms block both CD95 and TNF induced apoptosis
Apparently is recruited as a proenzyme to the receptor signaling complex by CD95-FADD and TNFR1- TRADD-FADD
Autoactivates in 2 steps: (1) abstraction of one of its two death effector domains (DED) into receptor complex may relieve autoinhibition caused by interaction between two DED domains, which would prime FLICE/MAH for (2) second trans-cleavage activation step, brought about by close proximity of other recruited pro-FLACH/MACH molecules. This would lead to release of active FLICE/MACH
FLT3
Class III receptor tyrosine kinase preferentially expressed on hematopoietic progenitor cells
Activated FLT3 activates signal transduction pathways involved in proliferation of pluripotent and progenitor cells
Internal tandem duplication reported in acute myeloid leukemia (20%), myelodysplasia (3%); D835 mutation present in AML (7%), myelodysplasia (3%), ALL (3%)
References: Archives 2005;129:1299
FMC7
Late B cell differentiation marker
Positive staining: mantle cell lymphoma, hairy cell leukemia, prolymphocytic leukemia
Negative staining: Chronic lymphocytic leukemia
fms
macrophage colony stimulating factor (M-CSF) receptor; also called CSF-1R, at 5q33-34
Promotes proliferation and differentiation of monocytes-macrophages
Mutations cause constitutive receptor transduction; associated with hematopoietic diseases, including myelodysplastic syndrome
Positive staining (normal): macrophages and their precursors
Positive staining (disease): choriocarcinoma, some AML
Focal adhesion kinase (FAK)
Protein tyrosine kinase, 125 kDa, that regulates antiapoptotic signaling
Binding of extracellular matrix molecules to integrins from endothelial cells causes recruitment of signaling molecules src, focal adhesion kinase, phospholipase C-gamma and basic-FGF to focal adhesion complexes, and modulates ability of endothelial cells to respond to growth factors
Positive staining (disease): invasive breast carcinoma (particularly high grade tumors, Mod Path 2005;18:1289), high grade sarcoma
Micro images: FAK+ breast carcinoma (fig 1a)
Fontana-Masson
Melanin stain; difficult to interpret faint staining in sparsely positive cells
Melanin granules reduce ammonia-silver nitrate and turn black
Micro images: melanoma
Formaldehyde induced fluorescence
Demonstrates catecholamines and indolamines
Biogenic amines plus formaldehyde vapors from heating form highly fluorescent derivatives
fos
Protein at 14q21-22 that binds DNA in complex with jun; an immediate early response gene
FRAP
FKBP Rapamycin Associated Protein
Also called mTOR or RAFT
Founding member of the phosphatidylinositol kinase-related kinase family
Sensor of physiological signals that regulate cell growth, including nutrients, cAMP levels, and osmotic stress
Affects transcription, translation, and autophagy.
Ability to mediate osmotic stress response may be via an intermediate mitochondrial dysfunction, Proc Natl Acad Sci USA 2002;99:4319
Fusin
T lymphocyte surface protein and key attachment site for HIV; works with CD4
Receptors that promote exchange of GTP for GDP bound to G alpha subunit, then dissociation of alpha GTP
and G beta gamma from receptor and each other; then alpha GTP or G beta gamma regulate effector protein; then GTP becomes GDP and alpha GDP reassociates with G beta gamma to turn off the signal
Galectin-3
Member of carbohydrate-binding protein family known as lectins
One of 14 galectins, which function as cell receptors for N-acetyl-lactosamine moieties present on most extracellular matrix components)
Also member of the beta-galactoside-binding protein family that plays an important role in cell-cell adhesion, cell-matrix interactions, macrophage activation, angiogenesis, metastasis, apoptosis
Uses: in one study, Gal-3+ with Ki-67 > 6% was associated with parathyroid carcinomas vs. adenomas (Hum Path 2005;36:908)
Positive staining (normal): endothelial cells, peripheral nerve, folliculostellate cells of adenohypophysis
Positive staining (disease): tumors of thyroid, head and neck, liver, colon, prolactinomas; parathyroid carcinoma; rarely in reactive of hyperplastic parathyroid lesions
Micro images: galectin-3 staining in lung squamous and adenocarcinoma
Reference: Mod Path 2005;18:1264 (prognostic significance in lung squamous cell and adenocarcinoma)
Gangliosides
GM1: on intestinal epithelial cell surface; bound by B unit of choleragen (Vibrio cholera toxin)
Gastrin releasing peptide
Appear at weak 15 of gestation
Relatively specific to neoplastic and non-neoplastic endocrine cells of the lung
Gelatinase B
See MMP-9
Gemori methamine silver
See GMS
Giemsa stain
As a hematology stain, works best with alcohol fixed smears
As a histology stain, detects mast cells and microorganisms, such as Giardia or Helicobacter
A "Romanowsky-type" stain, composed of mixtures of methylene blue, azure, and eosin compounds
Methylene blue is a metachromatic stain, meaning that some tissue components (mast cell granules ,cartilage, mucin, amyloid) stain purple and not blue
Glial fibrillary acidic protein (GFAP)
Intermediate filament for astrocytes (normal, reactive, neoplastic)
Positive staining (disease): CNS tumors, colonic schwannoma (AJSP 2001;25:846)
Micro images: myxopapillary ependymoma of broad ligament (fig 8b)
GLUT1
Facilitative glucose transporter; activated by hypoxia-sensing cellular pathways; may sustain cellular metabolism via glycolysis when hypoxia is present
Positive staining (normal): red blood cells, blood-brain barrier, perineurium
Positive staining (disease): various carcinomas, including fallopian tube carcinomas (Archives 2005;129:651)
Negative staining: benign epithelium
GLUT-4
Glucose transporter 4
Highly expressed in mitochondria-rich (oxyphil) cells in normal/neoplastic tissue, including gastric parietal cells, Hashimoto thyroiditis, Hurthle cell adenomas and carcinomas, occasional oxyphil parathyroid hyperplasias, occasional oxyphil parathyroid adenomas, renal oncocytomas, salivary gland oncycytomas, Warthin tumors, hibernomas
Glycophorin A
Also called CD235a
Positive staining: erythroid cells, AML-M6
Negative staining: AML M0-M5, M7
Glycosaminoglycans
Heteropolysaccharides which contain an N-acetylated hexosamine in a characteristic repeating disaccharide unit involving alternate 1,4- and 1,3-linkages consisting of either N-acetylglucosamine or N-acetylgalactosamine
GMS
Gomori methenamine silver
Special stain for detecting fungi and Pneumocystis carini
Stains cell walls and outlines these organisms; may have artefactual background staining
Micro images: Pneumocystis carinii; Cryptococcus neoformans; Coccidioides immitis; Aspergillus fumigatus
Gram stain
Stain to detect and differentiate bacteria
Method:
Apply crystal violet, then iodine, then decolorize by alcohol/acetone, then counterstain by safranin/fuchsin
Gram positive bacteria retain the crystal violet-iodine complex after decolorization, are not counterstained, and appear purple
Gram negative bacteria have a different cell wall structure, don’t retain the crystal violet-iodine complex after decolorization, and so are counterstained by safranin/fuchsin and appear pink/red
Micro images: various organisms #1; #2
Paraffin sections:
Use neutral red instead of safranin; gram negative organisms usually stain poorly because their bacterial wall lipid is removed in tissue processing
Note: with hematoxylin and eosin staining on paraffin sections, bacteria appear as blue rods or cocci regardless of gram reaction; colonies appear as fuzzy blue clusters
Rapid diagnostic strategy for bronchioalveolar lavage samples consists of Gram stain and bacterial ATP assay (Archives 2005;129:78)
Not suitable for burn wound surfaces (Archives 2003;127:1485)
Micro images:
Fungi - Fusarium in peritoneal fluid
Gram positive tissue sections - Bacillus anthracis in CSF; Lactobacillus osteomyelitis; methicillin resistant Staphylococcus aureus (MRSA) pneumonia; Mycobacterium abscessus (soft tissue); Nocardia asteroides
References: J Clin Pathol 1979;32:187, University of Utah method, University of Nottingham method
Granzyme B
Enzyme associated with cytotoxic T lymphocytes; induces apoptosis in target cells of these lymphocytes
Grimelius
Micro images: breast carcinoma - focal Grimelius+ cells in neuroendocrine DCIS; infiltrating ductal carcinoma with focal Grimelius+ cells; Grimelius stain+ colloid carcinoma #1; #2
Gross cystic disease fluid protein 15 (GCDFP-15)
Glycoprotein originally isolated in human breast gross cystic fluid
Positive staining (disease): lobular breast carcinoma (90%), primary breast carcinomas (72%), metastatic breast carcinoma (80%); also salivary gland and prostate carcinoma, apocrine differentiation
Micro images: breast carcinoma metastatic to stomach (fig 3c); neuroendocrine carcinoma of breast #1; #2
Growth factor receptors
Typically have extracellular ligand binding domain, a transmembrane domain and a cytoplasmic domain with a tyrosine kinase domain
Alterations (amplification, mutation, rearrangement) may lead to constitutive activation in absence of ligand.
gstp1
Gene directs formation of glutathione S transferase protein, which detoxifies environmental carcinogens by reduction
Inactivated by hypermethylation
Kidney tumors: stain must have pH between 1.5 and 2.0
Clear cell and papillary renal carcinoma have focal, coarse, droplike staining
Note: hemosiderin in any tumors will also stain positive
Uses: Helpful in distinguishing chromophobe renal cell carcinoma (intensely positive in large percentage of cells with reticular staining) from oncocytoma (usually negative; if positive - fewer cells with less intensity and dustlike staining)
HAM 56
Stains histiocytes, endothelium, adenocarcinoma
Negative staining: osteoclast-like giant cells
Micro images: giant cell tumor of bone - mononuclear cells are positive (fig 1B); giant cell tumor of tendon sheath - mononuclear and giant cells are positive (fig 2B)
Also called TSC1
On 9q34
Mutations cause tuberous sclerosis, an autosomal dominant disease beginning in infancy or early childhood with mental retardation and seizures, angiomyolipomas, subependymal giant cell tumors, cutaneous angiofibromas, cardiac rhabdomyomas, lymphangioleiomyomatosis and multifocal multinodular pneumocyte hyperplasia
Broadly expressed in many organs and tissues, including myometrium and most smooth muscle
Negative regulator of cell cycle – inhibits cell proliferation
Inactivation causes benign neoplasms in patients with tuberous sclerosis complex
HBME
Marker of mesothelial cells, named after laboratory of Dr. Hector Battifora and MEsothelioma
Also positive in various thyroid carcinomas
hc2
Positive in hairy cell leukemia, activated B and T cells, plasma cells
hcg
human chorionic gonadotrophin
Glycoprotein with alpha and beta subunits, used to detect pregnancy
Positive staining (disease): choriocarcinoma, syncytiotrophoblast cells in other tumors
Negative staining: exaggerated placental sites, placental site nodules, placental site trophoblastic tumors, epithelioid trophoblastic tumors
Heat shock proteins
See hsp
Hemosiderin
Hemoglobin breakdown product that contains iron
Heparanase
Expressed in metastatic gastric carcinoma to lymph nodes (95), primary gastric carcinoma (47%), not in normal gastric epithelial cells (Mod Path 2005;18:205)
Degrades heparan sulfate, a principal component of basement membranes, functional receptor complexes that facilitate signal transduction, and the extracellular matrix
Hepatocyte growth factor (HGF)
Only ligand for c-MET
Pleiotropic growth factor that promotes cell proliferation, motility and morphogenesis; also has important roles in normal development, organ regeneration and tumor development
Positive staining (disease): melanoma, melanocytic lesions
Hep Par1 antibody
Hepatocyte Paraffin 1; Also called Hep
Recognizes mitochondrial antigen of hepatocytes
Highly sensitive (92%); negative in higher nuclear grade tumors, AJSP 2002;26:978
Moderately specific; false positive cases were CK7+ or CK20+ (adenocarcinoma), chromogranin+ or synaptophysin+ (neuroendocrine)
Interpretation: granular cytoplasmic staining
Uses: determine hepatocellular origin, particularly in panel with alpha-fetoprotein and CEA or CD10 (canalicular pattern, more specific than Hep Par1)
Positive staining (disease): most hepatocellular carcinomas, some nonhepatocellular carcinomas metastatic to liver
Negative staining: bile duct adenoma
Micro images: highly malignant adult hepatic blastoma - fig A: ferritin; B: alpha-fetoprotein; C: HepPar1; D: CK8; E: p53; F: beta-catenin
HER2
Also called neu, c-erbB2, p185HER2
Proto-oncogene on #17q11-21 for human epidermal growth factor receptor 2
Member of HER/erbB family, which encodes 185 kDa transmembrane tyrosine kinase receptor proteins (also epidermal growth factor receptor, HER3 and HER4), whose activation causes a cascade of intracellular enzymatic pathways, which can cause cell transformation
Overexpression is an independent adverse prognostic factor in several cancers; usually attributed to amplification of HER2 gene
Tumor expression of HER2 is discordant at nuclear, cytoplasm and cell surface levels, which highlights limitations of immunohistochemistry alone (AJSP 2005;29:1125, pancreatic adenocarcinoma)
HER2 - Breast
Expression is regulated by transcription activation in normal breast
Protein overexpression is associated with gene amplification (at 17q21); note - chromosome 17 polysomy without HER2 amplification doesn’t appear to affect HER2 expression (AJSP 2005;29:1221)
Overexpressed in 20-30% of female breast cancers; associated with comedocarcinoma, aggressive tumors, also intraductal tumor spreading (Hum Path 2002;33:21); usually not due to chromosome 17 aneusomy (Mod Path 2002;15:137)
Amplification causes 90% of cases of HER2 overexpression
Amplification determines eligibility for Herceptin (trastuzumab - anti-HER2 antibody) treatment for breast cancer, but only weak / variable association with poorer prognosis /survival) or response to chemotherapy
85% concordance between immunohistochemistry and FISH in breast carcinoma
HER2 is often amplified in ADH and DCIS, suggesting involvement in initiation of breast carcinogenesis (Mod Path 2002;15:116)
Associated with c-myc amplification (Hum Path 2005;36:634)
10% of ER+ DCIS cases were also HER2+ and were high nuclear grade (Mod Path 2005;18:615)
Vulvar Paget’s disease: overexpression is common in primary and recurrent disease (Mod Path 2005;18:354)
Predominantly determined using immunostaining; relevant staining (3+) is strong, complete membranous staining of tumor cells evident at low power; can be confirmed by Fluorescence In Situ Hybridization (FISH, more sensitive but more expensive and difficult to distinguish in situ from invasive lesions) and Chromogenic In Situ Hybridization (CISH, Mod Path 2002;15:657, Mod Path 2005;18:1015)
Compare intensity to 3+ control slide with negative normal epithelium
For node negative patients, FISH and IHC results were generally similar with some discrepant cases (Archives 2001;125:746)
FDA approved assays give comparable results when strictly handled (Archives 2004;128:627)
Staining pattern:
0 (negative) - no staining or membrane staining in <10% of tumor cells