and Seema Pasha Apichai3
(1)
University of Florida, College of Medicine, Gainesville, FL, USA
(2)
Private Practice:, Orlando, FL, USA
(3)
Private Practice:, Chicago, IL, USA
8.1 Stains
Table 8-1:
Histologic Stains For Specific Tissue
|
Special Stain |
Target |
Color |
|---|---|---|
|
Collagen/Muscle | ||
|
Masson trichrome |
Collagen |
Blue or green |
|
Muscle, nerves, nuclei |
Dark red | |
|
Verhoeff-van Gieson |
Collagen |
Red |
|
Nuclei, muscles and nerves |
Yellow | |
|
Aldehyde fuchsin (Gomori) |
Collagen |
Red |
|
Mallory triple stain or trichrome stain (aniline blue) |
Collagen |
Blue |
|
Muscle |
Red | |
|
PTAH (phosphotungstic acid hematoxylin) |
Collagen |
Red |
|
Muscle/fibrin |
Blue | |
|
Movat’s pentachrome |
Collagen |
Yellow |
|
Muscle/fibrin |
Red | |
|
Elastic Tissue | ||
|
Verhoeff-van Gieson |
Elastic fibers |
Black |
|
Orcein-Giemsa (Pinkus acid orcein) |
Elastic fibers |
Black |
|
Aldehyde fuchsin (Gomori) |
Elastic fibers |
Blue or purple |
|
Movat’s pentachrome |
Elastic fibers |
Black |
|
Nerve | ||
|
Bodian |
Nerve fibers |
Black |
|
Fat | ||
|
Oil-red-O |
Lipid |
Red |
|
Sudan black B |
Lipid |
Black |
|
Scarlet red |
Lipid |
Red brown |
|
Melanin | ||
|
Fontana-Masson |
Melanin |
Black |
|
Orcein-Giemsa |
Melanin |
Brown-black |
|
Silver nitrate |
Melanin |
Black |
|
Hemosiderin | ||
|
Perls |
Hemosiderin/iron (not melanin) |
Bright blue |
|
Prussian blue |
Iron/hemosiderin |
Blue |
|
Turnbull’s blue |
Iron/hemosiderin |
Blue |
|
Calcium | ||
|
Alizarin Red S |
Ca++ |
Reddish orange |
|
Von Kossa |
Ca++ |
Brown black |
|
Mucin/Mucopolysaccharides | ||
|
Toluidine blue |
Acid MPS |
Purple |
|
Alcian blue |
Acid MPS (pH 2.5)
Sulfated MPS (pH 0.5) |
Blue |
|
Aldehyde fuchsin (Gomori) |
Acid MPS, elastic tissue |
Blue |
|
Colloidal iron |
Acid MPS |
Blue |
|
Crystal violet |
Acid MPS |
Purple with blue background |
|
Mucicarmine |
“Epithelial” mucin |
Bright pink ( Paget’s , cryptococcus capsule) |
|
Giemsa |
Acid MPS |
Metachromatically purple |
|
PAS |
Neutral MPS |
Pink |
|
Amyloid | ||
|
Congo red |
Amyloid |
Pink-red, green birefringence with polarization |
|
Crystal violet |
Amyloid |
Purple with blue background |
|
Thioflavin T |
Amyloid |
Yellow fluorescence under fluorescent microscope |
|
Scarlet red |
Amyloid |
Red |
|
Orcein-Giemsa |
Amyloid |
Light blue |
|
Mast cells | ||
|
Leder (chloroacetate esterase) |
Granules |
Red |
|
Giemsa |
Granules |
Purple |
|
Toluidine blue |
Granules |
Purple |
|
Tryptase |
Granules |
Red to brown |
|
Bacteria | ||
|
Gram stain |
Gram positive |
Blue
Brown-Brenn and Brown-Hopps – modified gram stain |
|
Gram negative |
Red | |
|
Fite |
M. leprae, Nocardia |
Red
Fite better than Ziehl-Neelsen for more delicate M. leprae bacilli |
|
Ziehl-Neelsen |
Acid fast bacteria |
Red |
|
Warthin-Starry |
Spirochetes |
Black |
|
Steiner |
Spirochetes |
Black |
|
Giemsa |
Leishmania, Histoplasma, Rickettsia |
Purple to blue |
|
DNA/RNA | ||
|
Methyl green-pyronin |
RNA |
Pink-red |
|
DNA |
Blue-green | |
|
Feulgen |
DNA only |
Magenta |
|
Fungi | ||
|
GMS (Gomori methenamine silver) |
Donovan bodies, fungi |
Black |
|
PAS (Periodic acid-Schiff) |
Fungi, neutral MPS, glycogen |
Red |
|
Grocott |
Fungi |
Fungus cell wall: black |
Table 8-2:
Immunohistochemical Stains
|
Marker |
Normal Location |
Positive in |
|---|---|---|
|
Epithelial markers | ||
|
Cytokeratin (Keratin) |
Epithelial cells, ± sweat glands
Types of keratin: AE1 (low molecular weight): basal epidermis, sweat glandsAE3 (high molecular weight): mid to superficial epidermisCAM 5.2: Paget’s disease; CK 7: Paget’s disease; CK 20: Merkel cell carcinoma |
Epithelial tumors, some adnexal tumors |
|
Melanocytic | ||
|
S100 |
Melanocytes, neural cells, smooth/skeletal muscle cells, Langerhans cells, eccrine and apocrine glands, chondrocytes |
Langerhans cell histiocytosis, melanoma, granular cell tumor, eccrine neoplasms, neural tumors, liposarcoma |
|
HMB-45 |
Melanocytes (less sensitive but more specific than S100) |
Melanoma, some normal nevi, Spitz nevus, angiomyolipoma, breast carcinoma |
|
Of note, MART-1/Melan A and Mel-5 comparable to HMB-45 | ||
|
Adnexal | ||
|
CEA (Carcinoembryonic antigen) |
Eccrine glands |
Paget’s disease, extramammary Paget’s, epithelioid sarcoma (sometimes), sweat gland tumors, adenocarcinomas (gastric, lung, breast, etc) |
|
EMA (Epithelial membrane antigen) |
Eccrine glands, some epithelial malignancies |
Paget’s disease, merkel cell carcinoma, meningioma, epithelioid sarcoma, most epithelial tumors |
|
Muscle | ||
|
Desmin |
Muscle cells (skeletal and smooth muscle) |
Leiomyoma, leiomyosarcoma, glomus cell tumor (focally +), embryonal rhabdomyosarcoma |
|
Vimentin |
Muscle cells, fibroblasts, endothelial cells, histiocytes, lymphocytes, Schwann cells, melanocytes |
Mesenchymal tumors (sarcomas, lymphomas, atypical fibroxanthoma, dermatofibrosarcoma protuberans), melanoma, glomus cell tumor |
|
Smooth muscle actin |
Myofibroblasts, muscle cells |
Glomus tumors, smooth muscle tumors, some atypical fibroxanthomas |
|
Neural | ||
|
Neuron specific enolase (NSE) |
Nonspecific neuroendocrine marker |
Granular cell tumor, merkel cell carcinoma, other neuroendocrine tumors, some melanocytic tumors (melanoma), schwannoma |
|
Glial fibrillary acidic protein (GFAP) |
Glial cells, astrocytes, Schwann cells |
|
|
Chromogranin |
Neuroendocrine neoplasms |
Merkel cell carcinoma |
|
Synaptophysin |
Neuroendocrine neoplasms |
Merkel cell carcinoma |
|
Myelin basic protein (MBP) |
Myelin sheath tissue, Schwann cells |
Neurofibroma, neuroma, granular cell tumor |
|
Endothelial | ||
|
Factor VIII related antigen (von Willebrand) |
Endothelial cells (not as sensitive as CD31) |
Vascular tumors (both benign and malignant) |
|
CD31 |
Monocytes, granulocytes, T/B cells, endothelial cells |
Vascular neoplasms (angiosarcoma) |
|
Ulex europaeus agglutinin 1 (UEA) |
Endothelial cells, most eccrine glands, keratinocytes |
Vascular tumors |
|
CD34 |
Benign and malignant vascular tumors, endothelial cells, nerves, hematopoietic cells |
Vascular tumors (benign/malignant), dermatofibrosarcoma protuberans, vascular tumors (not as sensitivie as ulex), neurofibroma, epithelioid sarcoma, spindle cell lipoma, fibrous papule |
|
Miscellaneous | ||
|
Factor XIIIa |
Macrophages, dermal dendrocytes, platelets |
Dermatofibroma, xanthoma disseminatum, fibrous papule, AFX, xanthogranuloma |
|
Bcl-2 |
Indicator of ↓ apoptosis, ↑ cell longevity |
Follicular center cell lymphoma |
|
Gross Cystic Disease Fluid Protein 15 (GCDFP-15) |
Apocrine glands |
Apocrine tumors, breast carcinoma, salivary gland tumors |
Table 8-3:
Specific Cells and Markers
|
Cell type |
Markers |
|---|---|
|
Macrophage |
α1-antitrypsin, lysozyme, HAM 56, CD 68 |
|
Melanocyte |
S100, HMB 45, mart1/melanA, vimentin |
|
Langerhans cell |
S100, CD1a, vimentin |
|
Merkel cell |
CK20, neurofilament, chromogranin A, NSE, synaptophysin, ± EMA |
|
Sebaceous gland |
EMA, keratin |
|
Sweat gland |
CEA, EMA, keratin (cytoplasmic positivity), gross cystic disease fluid protein-15 (in apocrine and some eccrine), smooth muscle actin (in myoepithelial cells) |
|
Fibroblast |
Vimentin |
|
Mast cell |
CD117 (c-kit) |
|
Myofibroblast |
Muscle specific actin, desmin (sometimes) |
|
Dermal dendrocytes |
Factor XIIIa |
|
Blood vessels |
Factor VIII-related Ag, CD31, CD34, vimetin |
|
Nerves |
Axons: neurofilament, NSE
Schwann cells: S100, GFAP, myelin basic protein (MBP) |
|
Muscle |
Smooth muscle actin |
Table 8-4:
Specific Tumors and Respective Markers
|
Entity |
Positive Markers: |
Negative Markers: |
|---|---|---|
|
Atypical fibroxanthoma (AFX) |
Vimentin, actin, + histiocyte stains (i.e. CD68) |
S100 (positive in spindle cell melanoma), pankeratin (positive in spindle cell SCC), CD34, CEA, EMA, desmin |
|
Leiomyosarcoma |
Desmin, smooth muscle actin, ± vimentin |
CEA, S100 (only rarely positive), keratin (only rarely positive) |
|
Melanoma |
S100, vimentin, melanA, HMB45 (latter negative in desmoplastic/spindle cell melanoma) |
Keratin (positive in Paget’s), CD45 (positive for lymphocytes in halo nevus), CEA, EMA, desmin |
|
Angiosarcoma |
CD31, CD34, Factor VIII-related antigen, Ulex europaeus |
Keratin, CEA, EMA, S100 |
|
Merkel cell carcinoma (MCC) |
Cytokeratin 20, EMA, NSE, synaptophysin, chromogranin |
S100, LCA, CEA, vimentin, desmin, GFAP, thyroid transcription factor-1 or TTF-1 (positive in small cell lung cancer) |
|
Schwannoma (Neurilemmoma) |
Vimentin, S100, MBP |
Keratin, neurofilaments, desmin |
|
Glomus tumor |
Vimentin, smooth muscle actin |
Desmin (occasionally focally positive), cytokeratin |
|
Epithelioid sarcoma |
Cytokeratin, CD34 (50 % cases), EMA, vimentin, ± CEA |
|
|
DFSP |
Vimentin, CD34 |
S100, HMB45, keratin, stromelysin-3,
Factor XIIIa (latter two markers positive in dermatofibroma) |
|
Paget’s disease |
CK7, EMA, CEA, GCDFP-15 |
S100, HMB45 |
|
Sebaceous carcinoma |
EMA, keratin (CAM 5.2) |
CEA (usually), S100 |
|
Granular cell tumor |
S100, NSE, MBP |
GFAP, neurofilament |
Table 8-5:
Markers for Different Tumors
|
Tumor |
Keratin |
S100 |
Vimentin |
Actin |
CD31 |
CD34 |
CD45 |
CD68 |
|---|---|---|---|---|---|---|---|---|
|
Carcinoma |
+ |
− |
− |
− |
− |
− |
− |
− |
|
Fibrohistiocytic |
− |
− |
+ |
+/− |
− |
+/− |
− |
+ |
|
Sarcoma |
− |
− |
+ |
+/− |
+/− |
+/− |
− |
− |
|
Angiosarcoma |
− |
− |
+ |
− |
+ |
+ |
− |
− |
|
Melanoma |
− |
+ |
+ |
− |
− |
− |
− |
− |
|
Lymphoma |
− |
− |
± |
− |
− |
− |
+ |
− |
|
Muscle |
− |
− |
+ |
+ |
− |
− |
− |
− |
|
Nerve |
− |
+ |
+ |
− |
− |
+ |
− |
− |
Table 8-6:
Differential Diagnosis of Spindle Cell Tumors
|
Entity |
Keratin |
S100 |
Factor VIII |
HMB-45 |
Desmin |
Vimentin |
|---|---|---|---|---|---|---|
|
SCC
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