Pathology

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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Aug 7, 2017 | Posted by in Dermatology | Comments Off on Pathology
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