Elston Chapter 1 Basics Flashcards Preview

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Flashcards in Elston Chapter 1 Basics Deck (148):
1

Anaplasia

atypical nuclei and pleomorphism

2

Asteroid body

collections of eosinophilic material seen in sporotrichosis also refers to star-shaped intracytoplasmic inclusions seen in giant cells of sarcoidosis or berylliosis or other granulomatous process

3

ballooning degeneration

destruction of epidermis by dissolution of cell attachments and intracellular edema

4

caterpillar bodies

pale pink linear basement membrane material within epidermis seen in PCT, represents degenerated type IV collagen

5

civatte/colloid bodies

pink, globular remnants of keratinocytes (lichen planus)

6

cornoid lamellae

45 degree anle parakeratosis in a column above a focus with diminished granular layer and underlying dyskeratotic cells

7

corps ronds/grains/dyskeratosis

corps ronds =- rounded nucleus with ahlo of poale to pink dyskeratotic cytoplasm corps grains = dark blue flattened nucleus surrounded by minimal cytoplasm dyskeratosis = abnormal individual-cell keratinization

8

Cowdry A body

also known as Lipshutz body intranuclear pink inclusions of herpesvirus infections

9

Cowdry B body

intranuclear pink inclusions of adenovirus and poliovirus infection

10

Donovan body

intracytoplasmic collectinos of bacteria seen in granuloma inguinale

11

Dutcher body

intracytoplasmic pink masses of immunoglobulin that invaginate into the nucleus of plasma cells and appear to be intranuclear

12

epidermolytic hyperkeratosis

coarse, irregular hypergranulosis associated with disruption of cell membranes associated with keratin 1 and 10 mutations

13

festooning

papillary dermis retains undulating pattern (often in PCT)

14

flame figure

collagen encrusted with major basic protein from eosinophils

15

guarnieri body

eosinophilic inclusions of smallpox

16

henderson-paterson body

intracytoplasmic oval, pink inclusions of molluscum infection

17

hyper/hypopigmentation

increased/decreased melanin pigment

18

kamino body

dull pink to amphophilic basement membrane material within epidermis in Spitz

19

koilocytes

keratinocytes with clear cytoplasm and shrunken raisin-like pyknotic nuclei

20

medlar body

chromomycosis copper penny spores brown round structure resembling overlapping copper pennies divide by septation, resembling a hot-cross bun

21

michaelis-gutman body

intra-and-extracellular calcified, concentric circular structures, seen in malakoplakia

22

papillary mesenchymal body

structure resembling whorl of plump mesenchymal cells normally present in the hair papilla (seen in trichoblastoma and trichoepithelioma)

23

Psammoma body

extracellular laminated, calcified structures seen in meningioma, papillary thyroid carcinoma, ovarian carcinoma

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24

pseudoepitheliomatous hyperplasia

acanthosis of adnexal epithelium and epidermis mimcking SCC, often a/w trapping of elastic fibers

25

pseudohorn cyst

keratin-filled cystic structure result of cutting through invaginations of stratum corneum (connects to surface)

26

reticular degeneration

destruction of epidermis with cell membranes remaining in a net-like pattern

27

russell body

intracytoplasmic pink inclusions of fIg in plasma cells, seen in rhinoscleroma and other conditions with many plasma cells

A image thumb
28

Schaumann body

laminated calcified structure seen in sarcoidosis

A image thumb
29

verocay body

composed of 2 nuclear palisades enclosing pink cytoplasmic processes, seen in schwannoma

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30

eyelid and ear skin

have many vellus hair follicles; eyelid skin has skeletal muscle in upper dermis

conjunctival side of eyelid skin has no stratum corneum or hair follicles, does have goblet cells

31

Areolar skin

slight acanthosis with basilar hyperpigmentation

sometimes central invagination of epidermis that leads to follicle and sebaceous glands

smooth muscle bundles in mid-deep dermis

apocrine glands in the reticular dermis

32

acral skin

compact eosinophilic stratum corneum

slight papillomatosis on dorsal surfaces

33

volar skin

compact eosinophilic hyperkeratosis with underlying stratum lucidum

no hair follicles or sebaceous glands

eccrine glands numerous

Meissner and Pacinian corpuscles can be seen

34

Mucosa

absent granular layer

keratinocytes large and pale (filled with glycogen)

dilated vessels in submucosa

can get smooth muscle bundles

35

nasal turbinate

erectile tissue with fibrous septa and vascular sinusoids and mucous glands

36

fetal skin

stellate and spindled fibroblasts (mesenchyme)

densely cellular

37

Hair anatomy

infundibulum - from epidermis to insertion of sebaceous gland

isthmus - from insertion of sebaceous gland to insertion of arrector pili muscle (bulge)

stem - from insertion of arrectorp ili muscle (bulge) to Adamson's fringe (only present in anagen hairs)

Bulb - below the step of anagen hair follicle, from Adamson's fringe to base of hair follicle

38

hair infundibulum

from epidermis to insertion of sebaceous gland

intraepidermal portion = acrotrichium

keratinizes in pattern of normal epidermis with granular layer

39

hair isthmus

from insertion of sebaceous gland to insertion of arrector pili muscle (bulge)

keratin formed in ABSENCE of granular layer = trichilemmal keratinization

inner root sheath is lost at this level and outer rooth sheath develops an inner corrugated, dense pink cornified layer; peripheral palisading of the outer root sheath is seen

40

Adamson's fringe (Hair)

point above which hair cornifies

dermaotophytes only infect cornified hair above Adamson's fringe

above Adamson's fringe, Huxley's layer of the inner root sheath no longer has trichohyalin granules

hair tends to retract from the inner root sheath above Adamson's fringe

the inner root sheath is fused and blue-gray at this level and trichohyalin granules are not seen

the outer root sheath is composed of pink cells with peripheral palisading

41

Bulb

below stem portion/from Adamson's fringe to base of hair follicle

3 zones: matrix, supramatrix, keratogenous zone

matrix - from base to critical line (widest point of bulb and papillae)

supramatrix - from critical line to B-fringe (point at which outer root sheath becomes multilayered and Henle's layer no longer has trichohyaline granules)

keratogenous zone - from B-fringe to Adamson's fringe

layers of the hair follicle that can be seen: fibrous root sheath, vitreous basement membrane zone, outer rooth sheath, inner root sheath (henle's layer, huxley's layer, cuticle of inner root sheath), hair shaft (cuticle of hair shaft, cortex), medulla

42

anagen vs telogen hairs

anagen hairs have stem and bulb producing hair shaft

telogen hairs lack inferior segment

telogen hairs are easily recognized in vertical sections as the club hair and surrounding trichilemmal keratin give the imrpession of a flame thrower

43

eponychium

visible cuticle - thick keratinous material boarders proximal nail fold and adheres to nail plate

true cuticle is located beneath the visible portion and is derived from teh ventral proximal nail fold

44

hyponychium

everything ventral to nail plate

45

solehorn

subungual white to colorless keratin extending from ditsal nail bed underheath the onychodermal band to below the free,distal edge of the nail plate

46

types of keratinization of the nail

onychokeratinization (no granular layer)  ard keratin of nail plate

onycholemmal keratinization - ventral part of proximal nail fold (has granular layer), bed epithelium (cuticle, bed horny layer, solehorn) no granular layer

epidermoid keratinization - dorsal proximal nail fold, lateral folds, hyponychium

47

dermal dendrocytes

macrophage-type cell in dermis; many are factor 13a positive, some are S100 positive

likely serve as antigen presenting cells

48

giant cell types

foreign body - ncueli haphazard

Langerhans - nuclei in wreath shape

Osteoclast-like - ncueli haphazard and eccentric; cytoplasm deep pink with scallooped border that molds to adjacent cells

touton - ncuel in wreath with foamy cytoplasm peripherally

ringed siderophage - touton with hemosiderin (characteristic of fibrous histiocytoma type of DF)

49

histiocytes

epithelioid cell with central round/oval nucleus and surrounding cytoplasm

tend to coalsecein tissue without intervening connective tissue

phagocytosis and antigen presentation

50

Langerhans cell

dendritic cells in epidermis and dermis

CD1a+, S100+ peanut agglutinin, langerin +

reniform with eccentric nucleus

originate in bone marrow

contain birbeck granuels, tennis-racked-shaped rod and oval bodies seen in EM

51

Th1 vs Th2 cytokines

Th1 cells prdouce IL-1, IL-2, IL-12 and IFN-gamma and are important for cell-mediated immunity and function in activating macrophages

TH2 cells produce IL-4, IL-5 and IL-10 and are important for humoral immunity

52

what cytokine induces eosionphil production

IL-5

53

eosinophils contain

major basic protein, eosinophil cationic protein, catalase and other proteins

54

Masson's trichrome

collagen - blue green

smooth msucel - red

ex) scar (blue-green) vs leiomyoma (red)

pitfall: young collagen can stain red

55

Verhoeff-Van Gieson

stain elastic fibers black

ex) absence or reduction in scar, mid-dermal elastolysis, anetoderma, cutis laxa

ex) distorted fibers in PXE

get beaded elastic fibers in elastofibroma dorsei

56

toluidine blue

stains mast cell granules metachromatically

also stains mucin/acid mucopolysaccharides

can use in urticaria pigmentosa to get metachromatic staining

57

Leder stain

naphthol ASD choracetate esterase

mast cell cytoplasm stains red (not dependent on presence of granules)

also stains myeloid cells (eg leukemia cutis)

58

name 2 mast cell stains

toluidine blue

leder

59

Name 5 carbohydrate stains

PAS, alcian blue, colloidal iron, toluidine blue, mucicarmine

60

mucicarmine stains

acid mucopolysaccharides pink to red - stains mucinous capsule of Cryptococcus neoformans pink to red

61

colloidal iron stains

acid mucopolysaccharides blue

hyaluronidase digestion can be combined with colloidal iron to differentiate between hyaluronic acid and other mucosubstances

62

Alcian blue stains

acid mucopolysaccharides blue

in normal skin, most mucin is sulfated acid mucopolysaccharide (heparin, chondroitin, and dermatan sulfates)

in most patholgoic states with increased dermal mucin, mucin is prdominantely non-sulfated hyaluronic acid

non-sulfated acid mucopolysaccharides (hyaluoronic acid) stain with Alcian blue at pH 2.5 but not pH0.5

ex) follicular mucinosis, GA, myxoid cyst, dermal mucin in lupus

Sulfated acid mucopolysaccharides stain with Alcian blue at both pH 2.5 and pH 0.5

can use with or without hyaluronidase to differentiate hyalulronic acid from other mucopolysaccharides

63

PAS stains what

glycogen, neutral mucopolysaccharides (like BM), and fungi

glycogen is diastase labile, ie. sections exposed to diastase before staining do not stain red with PAS

useful for clear cell acanthoma and trichilemomma

fungai and neutral mucopolysaccharides (basement membrane) are diastase resistant

can see thickened vessel walls in porphyria

acid mucopolysaccharides (hyaluronic acid) do not stain with PAS

64

amyloid stains with what 3 stains

congo red - stains brick red and has apple green birefringence withp olarized light

thioflavin T - yellow to yellow-green with fluorescent microscope

Crystal violet - metachromatic stain for red-purple amyloid color

65

Iron stains with what?

Prussian blue (Perls stain) - ferric ions react to form a deep blue color

distinguish melanin from hemosiderin

does not ID iron in intact RBCs

66

Melanin stains with what

Fontana-Masson - silver stain resulting in black precipitate with melanin

67

calcium stains with what 2 stains?

von kossa - silver stain stains calcium salts black (PXE, calcinosis cutis, calciphylaxis)

Alizarin red - binds directly to calcium ions resulting in orange-red color

68

lipids stain with what stains?

oil red O

sudan black

osmium tetroxide

all require fresh tissue

 

69

bacteria stain with 

Brown Hopps - modification of Brown-Brenn technique; gram positive stains blue and gram negative stains red

70

Fungi stain with what 2 stains

PAS - PAS positive and diastase resistant

GMS (Grocott's methenamine silver) - gray-black reaction with fungal walls, also stains Nocardia and Actinomyces

71

Mycobacteria stain with what 4 stains?

Ziehl-Neelsen acid-fast tain

Fite acid fast stain

Kinyoun's acid fast stain

mycobacteria stain bright red

fite is preferred for partially acid-fast organisms such as lepra bacilli, atypical mycobacteria and Nocardia - it preserves color due to use of peanut oil and gentle decolorization

Auramine-rhodamine - mycobacteria fluoresce reddish yellow with fluorescent microscope

 

 

72

Spirochetes stain with what stains?

Warthin-Starry, Dieterle, Steiner (modeified Dieterle)

silver stains stain black spirochetes

ex) lyme (around vessels and in dermal papillae), syphilis (lower epidermis)

also stains Legionella, Bartonella and Donovan bodies of granuloma inguinale

73

Giemsa stains what organisms and features?

myeloid and mast cell granules purplish blue

bacteria, Leishmania and Histoplasma

74

AE1/AE3 stains what?

cocktail of higha nd low molecuar weight monoclonal cytokeratin antibodies

stains epdermis and adnexal epithelium

stains all epithelial tumors (SCC and adnexal)

also stains epithelioid sarcoma, synovial sarcoma and mesothelioma

 

75

pankeratin/CK polyclonal keratin is used rather than AE1/AE3 why?

greater sensitivity

76

p63 is expressed where

basal and spinous cells of epidermis

germinative cells of sebaceous glands

myoepthelial cells of sweat glands

NOT reactive in metastatic carcinoma -->helps differentiate from primary cutaneous adnexal neoplasms

reactive in cutaneous spindle cell SCC

77

CAM5.2 detects low molecular weight cytokeratines present in what?

glandular neoplasms

does not stain epidermis or stratified squamous epithelium

marks Paget's disease and extramammary Paget's

78

CK7 use for what?

dtermine origin of metastatic carcinoma

marker for non-GI adenocarcinoma

Mark's Paget's disease and extramammary Paget's

79

CK20 stains what tumor?

Merkel cell carcinoma - stains in paranuclear pattern and distinguishes from metastatic oat cell carcinoma of lung that is typically negative

used in determining origin of metastatic carcinoma

marker of rGI adenocarcinoma

highlights sparase Merkel cells within basaloid islands of desmoplastic trichoep but not BCC

80

CK7 +, CK20+

bladder, pancreatic

81

CK7+, CK20 -

breast, lung, mesothelioma

82

CK7-, CK20+

Colon cancer

83

CK7-, CK20-

hepatocellular, prostate, renal, neuroendocrine and SCC of lung

84

CK7 positive 

 

(generally non-GI adenocarcinoma)

bladder, pancreatic (CK20+)

breast, lung, mesothelioma (CK20-)

85

CK20 positive

(generally positive in GI adenocarcinoma)

bladder, pancreatic (CK7+)

colon (CK7-)

86

CDX2 stains what

intestinal adenocarcinoma

cutaneous metastatic colon adenocarcinoma and extrammamary Paget's a/w underlying colorectal tumor

87

RCC (renal cell carcinoma)

positive in most cutaneous mets from RCC and negative in other clear cell tumors of skin

88

TTF-1

thyroid transcription factor

reactive in metastatic small cell lung carcinoma but negative in Merkel cell carcinoma

89

EMA 

epithelial membrane antigen

highlights normal sebaceous and sweat glands

positive in sebaceous carcinoma, Paget's and extramammary Paget's

90

CEA

carincoembryonic antigen

highlights sweat glands, sweat gland neoplasms, Paget's, extrammamary Paget's and most adenocarcinomas

91

adipophylin stains

lipid droplets of sebaceous and xanthomatous lesions

helps distinguish sebaceous carcinoma from SCC and BCC

92

Ber-EP4 

marks most epithelial cells but not those with squamous differentiation

positive in BCC but negative in SCC

93

Desmin stains

sekeletal and most smooth muscle

negative in vascular smooth muscle including glomus cells

94

CD34 stains what normal structures

vascular endothelium and hematopoietic progenitor cells

positive in DFSP, negative in DF

positive in spindle cell lipoma, sclerotic fibroma, solitary fibrous tumor, superficial acral fibromyxoma, pleomorphic fibroma and pleomorphic hyalinizing angiectatic tumordecreased staining in morphea

increased staining in nephrogenic systemic fibrosis

stains connective tissue around normal hair folicles

typically hilights stroma of trichoeps but not BCCs

95

Factor 13a stains

DF, not DFSP

highlights dermal dendritic cells

positive in fibrous papule of hte face/angiofibroma

96

CD31 is a marker for

vascular origin of tumors

more specific for vascular than CD34

97

D2-40 (podoplanin) is a marker of

lymphatic endothelium

increases detection of lymphovascular invasion

lack of reactivity in metastatic carcinoma assiststs in differentiation from primary cutaneous adnexal neoplasms

98

UEA-1 *Ulex europeus agglutinin) binds 

vascular endothlium

99

GLUT1 STAINS

endothelial cells with blood-tissue barrier functiona sin placenta

positive in infantile hemangiomas and negative in vascular malformations

also stains perineurial cells and perineurioma

100

Vimentin stains

general marker of sarcomas

stains mesenchymal cells, endothelial cells, fibroblasts, melanocytes, lymphocytes, macrophages

does not react with epithelium/keratinocytes

excludes most carcinomas except rare spindle cell carcinomas and synovial carcinoma

101

S100 stains

neural crest-derived cells and some mesenchymal lines

staines melanocytes, Langerhans cells, sweat glands, nerves, schwann cells, myoepithelial cells, fat, muscle and chondrocytes

useful in ddx of spindle cell neoplasms

stains desmoplastic melanoma, langerhans cell histiocytosis, granular cell tumor, Rosai-Dorfman disease

102

S100A6 (calcyclin) stains

member of S100 superfamily

positive in cellular neurothekeoma while S100 is negative

reactive in most atypical fibroxanthomas but not specific

also stains other spindle cell tumors

stains Spitz strongly and diffusely but spitzoid melanomas have weak or patchy staining

103

HMB-45 is a marker of

premelanosome

loss of stianing of melanocytes with descent into dermis is manifestation of loss of premalnoosomes, marker ofr normal  maturation

loss of staining in deep dermal component of most benig nevi but get uniform staining of blue nevi

does not stain desmoplastic melanoma reliably

104

melan-A and Mart-1

2 different antibodfies stain same epitope

stain melanocytic lesions but staining of desmoplastic melanoma is unreliable

105

p75 (nerve growth factor receptor)

early neural crest cell marker

expressed in type C (spindled) melanocytes and schwann cells

sensitive marker for spindle cell and desmoplastic melanoma

106

MITF (micropthalmia-associated transcription factor)

essential in development and survival of melanocytes

nuclear melanocyte marker

positive in cellular neurothekeomas

107

Sox-10

nuclear marker of melanocytes and Schwann cells

sensitive marker of melanoma including conventional, spindled and desmoplastic

108

Neuron-specific enolase (NSE)

positive in neuroendocrine cells, neurons and tumors derived from them

fairly non-specific as positive in many other cell lines

109

Chromogranin positive in what tumor?

merkel cell carcinoma

110

synaptophysin positive in what tumor?

merkel cell carcinoma

111

CD45Ra (LCA)

leukocyte common antigen (LCA) general marker of hematolymphoid differentiation

present on all hematopoietic cells and precursors with exception of maturing erythroids and megakaryocytes

112

CD45Ro (UCHL-1)

stains mature T cells

113

CD20 stains

B cells (often absent in plasma cells)

positive in B cell lymphoma

target for rituximab

loss correlates with rituximab resistance

114

CD10 (CALLA)

common acute lymphoblastic leukemia antigen 

early marker of B cell differentiation

useful for diagnosis of B cell lymphoproliferative disorders

positive in periadnexal mesenchymal cells, staining only stroma of trichoblastomas but epithelial cells of BCC

expressed in most atypical fibroxanthomas but not often seen in other spindle cell tumors

marker of renal cell carcinoma but also in other clear cell lesions including balloon cell nevi, clear cell hidradenoma and sebaceous tumors

115

CD79a marker for 

plasma cell and B cell

116

CD138 (syndecan-1) stains

 plasma cell marker

117

CD3 stains

T cells - pan -T-cell marker

118

CD4 stains

helper T cells

119

CD8 stains

cytotoxic/suppressor T cell marker

120

CD5 stains

pan-T cell marker but aberrant loss in CTCL is common

positive in mantle cell lymphoma and infiltrates of chronic lymphocytic leukemia

121

CD30 (Ki-1, BERH2) stains

Reed-Sternberg cells of Hodgkin's disease

positive in activated lymphs of anaplastic large cell lymphoma and lymphomatoid papulosis

many positive cells may be seen in scabies nodules and chronic tick bites

122

CD7

immature T lymphocyte antigen

most common antigen lost in T cell lymphoma

123

CD56

marker of NK cells and subset of T cells

stains blastic plasmacytoid dendritic cell neoplasm (fomerly known as blastic NK/T-cell lymphoma or CD4+/CD56+ hematodermic neoplasm)

124

CD68 (KP-1)

reactive in virtually all monocyte/macrophage cells

125

CD163

reactive in monocytes and macrophages

126

CD123

marker of plasmacytoic dendritic cells

positive in blastic plasmacytoid dendritic cell neoplasm

127

myeloperoxidase

major constituent of granuleso f neutrophilic myeloid cells

marker for acute myeloid leukemia

128

ALK-1

anaplastic lymphoma kinase expressing chromosomal translocation t(2,5)

positive in most systemic anaplastic large cell lymphoma and negativ ein primary cutaneous anaplastic large cell lymphoma

few patients with ALK-1 negative systemic anaplastic large cell lymphoma have poor prognosis

129

Kappa/lambda

normally expressed in ratio f 2/3 kappa to 1/3 lambda

10 fold deviation suggests clonal B-cell proliferation

130

CD117 (c-Kit)

expressed in mast cells and melanocytes

in nevi and primary melanoma there is decrease in expression in dermal component

typically lost in metastatic cutaneous melanoma

131

CD1a

stains Langerhans cells

132

Langerin (CD207) stians

Birbeck granules in Langerhans cells

133

CD43 (Leu-22)

pan-T-cell marker

aberrant coexpression with CD20 strongly suggestive of B cell lymphoma

134

BCL2

onocogene that inhibits apoptosis

useful to differentiation B cell lymphoproliferative disorders

most BCCs have diffuse BCL2 staining wheras trichoeps only show staining of outermost epithelial layeres of tumor islands

135

MUM-1 (Multiple myelanoma oncogene-1)

expressed in plasma cells, activated T cells, and subset of germinal center cells

distinguishes primary cutaneous diffuse large B cell lymphoma, leg type from diffuse follicle center lymphoma

reactivity reported in anaplastic large cell lymphoma, LyP and blastic plasmacytoid dendritic cell neoplasm

also expressed in benign and malignant melanocytic cells

136

CD21

follicular dendritic cell marker

highlights residual follicle in lymphoma

CD23 has a similar staining pattern

137

betaF1

identified alphabeta T cells

138

Spindle cell neoplasms should be stianed with what?

pankeratin or AE1/AE3 - positive in SCC

CD10 - positive in atypical fibroxanthoma, may be positive in SCC, melanoma, leiomyosarcoma

S100/Sox-10 - positive in melanoma

Desmin - positive in leiomyosarcoma

139

small blue cell tumors should be stained with

LCA - positive in lymphoma

TTF-1 - positive in metastatic small cell carcinoma of lung, sometimes in lymphoma

CK20 - positive in Merkel cell carcinoma

Synaptophysin - positive in Merkel cell carcinoma, sometimes metatstatic small cell carcinoma of lung

S100 - positive in melanoma, sometimes merkel cell carcinoma

140

small round blue cell S100 positive

probably melanoma, could be merkel cell (get synaptophysin or CK20)

141

small round blue cell TTF-1 positive

probably metastatic small cell caricnoma of lung, could be lymphoma, get LCA (Positive in lymphoma)

142

small round blue tumor positive for synaptophysin

probably merkel cell but could be metastatic small cell carcinoma of the lung, get TTF-1 (positive in small cell carcinoma of lung)

143

What tumors have intraepidermal buckshot scatter (pagetoid spread)

Paget's, extramammary Paget's, Bowens, Malignant melanoma

144

for pagetoid scattered order what stains?

CK7 (positive in Paget's and sometimes Bowens disease)

CEA (positive in Paget's)

S100 (positive in melanoma)

145

specific immunohistochemical antibodies available to many viruses, bacterial, fungal and parasitic antigens including

HHV8, HSV, VZV, CMV, EBV, bartonella, Rickettsia, treponema, Borrelia, Aspergillus, Leishmania and others

146

markers of proliferation include 

Mib-1 (Ki-67) - nuclear prolifereation marker expressedin all active phases of cell cycle

pHH3 - mitotic marker only stains M phase of cell cycle; helps differentiate mitoses from apoptotic or hyperchromatic nuclei

147

what transport media should be used for electron microscopy

glutaraldehyde

148

what transport medium should be used for immunofluorescence?

Michel's medium (ammonium sulfate). if not available, use normal saline.