M3-MUSCLE AND BONE Flashcards
(203 cards)
1
Q
Type of muscle with voluntary control and striations
A
Skeletal muscle
2
Q
Characteristics of skeletal muscle fibers
A
Large elongated eosinophilic myofibers+Multiple peripheral nuclei+Striations at right angles to fiber axis
3
Q
Energy source abundant in skeletal muscle
A
Glycogen
4
Q
Type of muscle with involuntary control and spindle-shaped fibers
A
Smooth muscle
5
Q
Characteristics of smooth muscle fibers
A
Smaller tapered myofibers+Elongated central nuclei+Pale cytoplasm with hematoxylin
6
Q
Locations of smooth muscle
A
Gastrointestinal tract+Urogenital tract+Blood vessels
7
Q
Type of muscle found in heart with branching cells and intercalated discs
A
Cardiac muscle
8
Q
Characteristics of cardiac muscle cells
A
Branching cells+Intercalated discs+Centrally placed nuclei+Lightly stained nuclei with hematoxylin
9
Q
Three parts of open muscle biopsy
A
Small segment (glutaraldehyde for EM)+Middle segment (frozen for enzyme histochemistry)+Remaining segment (buffered formaldehyde for light microscopy and IHC)
10
Q
Common needle biopsy site for muscle
A
Thigh
11
Q
Staining method for glycogen in muscle
A
PAS + diastase
12
Q
Stains used for lipids in muscle
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Oil Red O+Sudan Black
13
Q
ATPase staining at pH 9.4 demonstrates
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Myosin loss and atrophy of Type 1 and 2 fibers
14
Q
ATPase staining at pH 4.6 demonstrates
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Type 2B myofibers
15
Q
ATPase staining at pH 4.3 demonstrates
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Type 2C myofibers
16
Q
Stain demonstrating internal fiber architecture and mitochondria
A
NADH-TR
17
Q
Stain for regenerating myofibers and autoimmune connective tissue disorders
A
Alkaline phosphatase
18
Q
Stains for inflammatory cells and necrotic myofibers
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Acid phosphatase+Non-specific esterase
19
Q
Stains for mitochondrial disorders
A
Cytochrome c oxidase+Succinic dehydrogenase
20
Q
Enzyme deficiency stain
A
Myoadenylate deaminase
21
Q
Stain for Type IV glycogenosis (McArdle’s disease)
A
Myophosphorylase
22
Q
Dystrophy-related immunohistochemical stains
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Dystrophin 1 2 and 3+Sarcoglycans+Dysferlin+Merosin+Caveolin+Emerin+Calpain-3+Spectrin
23
Q
Trichrome stain color for erythrocytes muscle and collagen
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Orange+Red+Blue
24
Q
Most common trichrome stain to differentiate muscle from collagen
A
Masson’s trichrome
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Masson’s trichrome staining sequence
Red dye overstain+Polyacid displacement+Second acid dye stains collagen blue or green
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Masson’s trichrome stain result in angiomylipoma
Smooth muscle red+Collagen blue
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Stain used for frozen sections in muscle biopsy
Gomori’s trichrome (chromotrope 2R fast green FCF phosphotungstic acid and glacial acetic acid)
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Use of Gomori’s trichrome for frozen muscle
Detecting ragged fibers in mitochondrial myopathy
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Color of myofibrils in Gomori’s trichrome stain
Green
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Color of intermyofibrillar material nemaline rods and ragged red fibers in Gomori’s trichrome
Bright red
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Function of phosphotungstic acid in Mallory’s PTAH
Binds tissue proteins and hematin forming a blue lake pigment
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Tissues demonstrated by Mallory’s PTAH
Striated muscle fibers and mitochondria
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Appearance of muscle in Mallory’s PTAH
Blue-black to dark brown
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Appearance of connective tissue in Mallory’s PTAH
Pale orange-pink to brownish red
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Appearance of fibrin in Mallory’s PTAH
Deep blue
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Appearance of coarse elastic fibers in Mallory’s PTAH
Purple
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Appearance of bone and cartilage in Mallory’s PTAH
Yellowish to brownish red
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Material located in intermyofibrillar network staining positive with PAS
Glycogen
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Muscle fiber type staining darker with PAS
Type 2 myofibers
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Use of PAS stain in muscle
Detect glycogen storage and visualize basement membrane of capillaries
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Issue with glycogen staining in paraffin or cryostat sections with PAS
Glycogen dissolves easily causing false negatives
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Color of intracellular lipid in Sudan Black stain
Blue-black
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Muscle fiber type staining darker with Sudan Black
Type 1 myofibers
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Use of Oil Red O stain
Staining neutral lipids for fluorescence or bright field microscopy
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What does Oil Red O stain
Neutral lipid forming part of intracellular membranes
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Association of fat accumulation
Mitochondrial myopathy
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Appearance of neutral lipids in Oil Red O stain
Red
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Appearance of nuclei in Oil Red O stain
Pale blue
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Tissues stained gray-black with Heidenhain’s iron hematoxylin
Muscle striations mitochondria myelin and chromatin
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Purpose of histochemical stains/enzyme histochemistry
Highlight selective fiber type involvement or myofiber architecture aspects
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Muscle fiber type for continual/postural functions with numerous mitochondria and lipid
Type I (slow twitch/slow-oxidative)
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Muscle fiber type for brief intense activity specialized for anaerobic metabolism
Type II (fast twitch)
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Staining of Type I fibers in ATPase reaction at pH 9.4
Light
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Staining of Type II fibers in ATPase reaction at pH 9.4
Dark
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What are Type II fibers rich in
Glycogen myophosphorylase phosphofructokinase and ATPase
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What are Type II fibers marked by
Sharp staining with myosin-ATPase and phosphorylase paucity of staining with oxidative enzyme techniques
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Which fibers Contains less mitochondria and triglyceride lipid
Type II Fibers
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Subtypes of Type II fibers
Type IIA Type IIB and Type IIC
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Type IIA fibers characteristics
Large number of mitochondria and myoglobin manufactures and splits ATP quickly using aerobic and anaerobic metabolism
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What are Type IIB fibers
White in color due to a low level of myoglobin and few mitochondria Produce ATP at a slow rate by anaerobic metabolism and break it down very quickly short fast bursts of power and rapid fatigue Muscle of the arms
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Reliable method to distinguish muscle fiber types
Adenosine triphosphatase (ATPase stain)
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What does the fibers are separated based on
Differences in pH sensitivity
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What do Employ solutions of different pH values primarily to distinguish between
Fast- and slow- contracting muscle fibers
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Describe pH 9.4 ATPase Fibers
Type 1 fibers light Type2 fibers dark Type 2C fibers intermediate
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Describe pH 4.6 ATPase Fibers
Type 1 fibers darkest Type 2B and C intermediate Type 2A lightest
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Describe pH 4.3 ATPase Fibers
Type 1 fibers darkest Type 2C fibers intermediate Type 2A and 2B fibers lightest
67
Succinate dehydrogenase (SDH) location
Inner mitochondrial membrane
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SDH function
Oxidation of succinate to fumarate in citric acid cycle
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Use of SDH stain
Distinguish oxidative (type 1) and less oxidative (type 2) muscle fibers
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SDH reaction product
Nitro blue tetrazolium (NBT) purple salt producing speckled pattern
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Appearance of oxidative fibers in SDH stain
Dense purple speckled
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Appearance of nonoxidative fibers in SDH stain
Scattered purple speckles
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SDH stain sensitivity
Most sensitive stain for detecting mitochondrial proliferation
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NADH-tetrazolium reductase (NADH-TR) stain target
Mitochondria and sarcoplasmic reticulum
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Reliability of NADH-TR as mitochondrial marker
Not reliable because stains sarcoplasmic reticulum too
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Use of SDH and COX stains
Better mitochondrial markers than NADH-TR
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Tubular aggregates stained by NADH-TR but not SDH
Tubular aggregates in sarcoplasmic reticulum
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Appearance of lobulated muscle fibers with mitochondrial accumulation
NADH-TR shows dark wedge-shaped peripheral accumulations
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COX stain use
Detect cytochrome oxidase deficiency
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COX staining pattern in normal muscle
More intense in type 1 fibers than type 2
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Alpha-glycerophosphate dehydrogenase (α-GPD) stain use
Distinguish fibers by anaerobic ATP generation capacity
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α-GPD enzyme function
Oxidizes glycerol-1-phosphate linked to NBT producing homogeneous reaction
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Myophosphorylase stain function
Detect phosphorylase activity acting on glucose-1-phosphate forming polysaccharides
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Color reaction of myophosphorylase stain
Negative yellow; short polysaccharide chains reddish; longer chains blue
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Disease with myophosphorylase deficiency
McArdle’s disease (GSD V)
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Effect of McArdle’s disease on glycogen
Excessive glycogen accumulation due to lack of phosphorylase activity
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PAS stain in McArdle’s disease
Intensely positive due to glycogen accumulation
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Good stain to identify macrophages
Nonspecific esterase
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Location positive for nonspecific esterase
Neuromuscular and myotendinous junctions
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Appearance of denervated fibers with nonspecific esterase
Excessively dark staining
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Muscle fibers stained by nonspecific esterase during necrosis
Positive
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Process of engulfment of muscle fibers
Myophagocytosis
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Stain for lysosomal vacuoles and abnormal lysosomal activity
Acid phosphatase
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Use of acid phosphatase in muscle
Identify macrophages in necrotic fibers and abnormal lysosomal activity
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Mechanism of acid phosphatase staining
Phosphate released by acid phosphatase forms lead phosphate precipitate converted to black lead sulfide
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Disease with increased acid phosphatase activity and vacuoles
Adult onset acid maltase deficiency
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Muscle biopsy showing central cores with acid phosphatase reactivity
Central core myopathy
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Muscle biopsy with abundant sarcoplasmic vacuoles containing acid phosphatase positive granules
Infant onset acid maltase deficiency
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Two types of bone tissue
Cortical (compact) bone+Trabecular (cancellous or spongy) bone
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Characteristics of cortical bone
Solid hard strong bone forming shafts of long bones and exterior surfaces of flat bones
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Characteristics of trabecular bone
Mesh of bone strands found in diaphysis epiphysis marrow cavities of long bones vertebrae and centers of flat bones
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Major components of bone
Bone collagen+Bone mineral+Bone cells
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Classification of bone based on collagen orientation
Woven bone+Lamellar (mature) bone
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Features of woven bone
Coarsely oriented thick loosely packed collagen fibers+High mineral content and osteocyte density+Irregular calcification+Weaker more elastic than lamellar bone
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Features of lamellar bone
Delicately packed collagen bundles organized in parallel or concentric layers+Lower mineral content and osteocyte density+Stronger and less flexible than woven bone
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Composition of bone mineral
Calcium phosphate combined with hydroxyl ions forming hydroxyapatite crystals
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Bone cells and functions
Osteoblasts (bone formation)+Osteocytes (mature bone cells incorporated into matrix)+Osteoclasts (bone resorption)
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Techniques for analyzing decalcified bone
Frozen sections+Paraffin sections+Transmission electron microscopy
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Techniques for analyzing mineralized bone
Frozen sections+Transmission electron microscopy+Scanning electron microscopy
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Stains used for decalcified bone
Hematoxylin and eosin+Masson’s trichrome+PAS+Van Gieson’s picrofuchsin
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Methods to counteract weak nuclear staining in decalcified bone
Hematoxylin freshly prepared+Progressive staining or reduced acid differentiation+Sodium bicarbonate immersion
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Stain based on deposition of thionin precipitate in lacunae and canaliculi
Schmorl’s picro-thionin method
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Stain demonstrating calcium salts
Alizarin Red S
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Appearance of calcium deposits with Alizarin Red S
Orange-red and birefringent
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Stain based on precipitation of silver ions reacting with phosphate replacing calcium
Von Kossa stain
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Visualization of Von Kossa stain
Metallic silver deposits reduced by strong light
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Bone marrow aspirate smear preparation
Expel aspirate into dish draw particles with Pasteur pipette place on slides and smear
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Alternative smear preparation
Place drop of aspirate on slide drain excess blood before smearing
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Fixative for bone marrow aspirate smears
Acetone-free absolute methanol
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Common stains for bone marrow aspirate
May-Grunwald Giemsa+Wright Giemsa (Romanowsky stains)
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Additional techniques on bone marrow aspirate slides
Cytochemistry (myeloperoxidase nonspecific esterases)+Immunohistochemistry+Fluorescent in situ hybridization
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Purpose of squash smear
Assess cellularity megakaryocyte numbers focal disease and fibrosis
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Characteristic of squash smear
Considerable cellular artifact
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Purpose of spread smear
Excellent detail of individual cell morphology
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Bone marrow trephine biopsy length
At least 2 cm
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Fixatives for bone marrow trephine
Neutral buffered formalin+Zinc formaldehyde+B5+Acetic acid-zinc-formalin+Isotonic buffered formalin+Bouin’s+Formaldehyde and glutaraldehyde
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Imprint preparation from trephine
Touch fresh unfixed core on slide or slide on core fixed and stained like aspirate smears
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Decalcification agents for bone marrow trephine
EDTA+Formic acid+Acetic acid+Picric acid+Nitric acid
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Section thickness for bone marrow trephine
2-3 micrometers
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Stains for bone marrow trephine
Hematoxylin and eosin+Van Gieson
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Plastic embedding benefits for bone marrow
Good cytological detail+No decalcification needed+Useful for metabolic bone disease and histochemical reactions
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Components of Romanowsky stains
Methylene blue/azure B+Eosin dissolved in acetone-free methanol
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Characteristic of Wright’s stain
Methylene blue polychromed by heating with sodium bicarbonate+Eosinates dissolved in absolute methyl alcohol+No fixation needed before staining
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Color of erythrocytes in Wright’s stain
Yellowish-red
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Color of polymorphonuclear nuclei and granules
Nucleus dark purple+Granules reddish-lilac
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Color of polymorphonuclear cytoplasm
Pale-pink
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Color of eosinophil nuclei and granules
Nuclei blue+Granules red to orange-red
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Color of eosinophil cytoplasm
Blue
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Color of basophil nucleus and granules
Nucleus purple to dark blue+Granules very dark purple
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Color of lymphocyte nuclei and cytoplasm
Nuclei dark purple+Cytoplasm sky blue
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Color of platelet granules
Violet to purple
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Use of Giemsa stain
Identify plasma cells mast cells lymphoid cells eosinophils distinguish myeloblasts and proerythroblasts
143
Color of bile pigments with Giemsa
Green
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Collagen muscle and bone stain color with Giemsa
Pale pink
145
Micro-organisms (fungi parasites) stain color with Giemsa
Purplish-blue
146
Starch granules and cellulose stain color with Giemsa
Sky blue
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Nuclei stain color with Giemsa
Dark blue to violet
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Erythrocytes stain color with Giemsa
Salmon pink
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Cytoplasm stain color with Giemsa
Varying light blue shades
150
Nuclei stain color with Wright-Giemsa or Jenner-Giemsa
Purple blue
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Cytoplasm stain color with Wright-Giemsa or Jenner-Giemsa
Pink blue
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Eosinophils stain color with Wright-Giemsa or Jenner-Giemsa
Pink red
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Use of May-Grunwald-Giemsa stain
Define cytologic dysplasia in bone marrow aspirate samples (hallmark of myelodysplastic syndromes)
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Gold standard stain for detection of hemosiderin in bone marrow aspirate
Perls’ Prussian blue iron stain
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Principle of Perls’ Prussian blue iron stain
Ferric ions released by dilute hydrochloric acid react with potassium ferrocyanide to produce insoluble blue compound
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Conditions detected by Perls’ Prussian blue iron stain
Anemia+Iron overload+Myelodysplasia
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Reliability of core biopsy sections for storage iron assessment
Less reliable due to decalcification removing storage iron
158
Use of myeloperoxidase stain
Identify cytoplasmic granules of myeloid cells and differentiate myeloid leukemia from lymphoid origin
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Principle of myeloperoxidase stain
Iron-porphyrin enzyme promotes oxidation of benzidine by hydrogen peroxide forming blue-green compound with copper sulfate
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Myeloid cells staining with myeloperoxidase
Positive with green to dark blue granules
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Eosinophils staining with myeloperoxidase
Most intense brown-black or green-black granules
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Neutrophil cytoplasm staining with myeloperoxidase
Blue
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Basophils lymphocytes and erythroblasts staining with myeloperoxidase
Negative
164
Monocytes staining with myeloperoxidase
Slight peroxidase activity
165
Bone marrow section embedding for Masson’s trichrome
Undecalcified glycol methacrylate embedded sections cut at 4 micrometers
166
Stains in Masson’s trichrome for bone marrow
Osteoid seams red+Mineralized bone blue+Nuclei dark gray
167
Most useful stain for plastic-embedded tissue sections
Toluidine blue
168
Use of reticulin stain
Detection of myelofibrosis+Leukemia+Lymphoma
169
Ancillary procedures in bone marrow analysis
Immunohistochemistry+In-situ hybridization+Cytogenetics+PCR and gene sequencing
170
Types of lymph node biopsies
Fine needle aspiration+Open (excisional) biopsy+Sentinel lymph node biopsy
171
Processing methods for lymph node biopsies
Microscopy on fixed stained tissue+Immunohistochemistry and flow cytometry+Cytogenetic G-banding+FISH+Molecular genetic analysis PCR and sequencing
172
Use of cytogenetic or FISH analysis
Diagnosis of Burkitt’s lymphoma
173
Use of PCR in lymph node biopsies
Detection of specific translocations and T/B cell clonality
174
Collagen muscle and bone stain color with Giemsa
Pale pink
175
Micro-organisms (fungi parasites) stain color with Giemsa
Purplish-blue
176
Starch granules and cellulose stain color with Giemsa
Sky blue
177
Nuclei stain color with Giemsa
Dark blue to violet
178
Erythrocytes stain color with Giemsa
Salmon pink
179
Cytoplasm stain color with Giemsa
Varying light blue shades
180
Nuclei stain color with Wright-Giemsa or Jenner-Giemsa
Purple blue
181
Cytoplasm stain color with Wright-Giemsa or Jenner-Giemsa
Pink blue
182
Eosinophils stain color with Wright-Giemsa or Jenner-Giemsa
Pink red
183
Use of May-Grunwald-Giemsa stain
Define cytologic dysplasia in bone marrow aspirate samples (hallmark of myelodysplastic syndromes)
184
Gold standard stain for detection of hemosiderin in bone marrow aspirate
Perls’ Prussian blue iron stain
185
Principle of Perls’ Prussian blue iron stain
Ferric ions released by dilute hydrochloric acid react with potassium ferrocyanide to produce insoluble blue compound
186
Conditions detected by Perls’ Prussian blue iron stain
Anemia+Iron overload+Myelodysplasia
187
Reliability of core biopsy sections for storage iron assessment
Less reliable due to decalcification removing storage iron
188
Use of myeloperoxidase stain
Identify cytoplasmic granules of myeloid cells and differentiate myeloid leukemia from lymphoid origin
189
Principle of myeloperoxidase stain
Iron-porphyrin enzyme promotes oxidation of benzidine by hydrogen peroxide forming blue-green compound with copper sulfate
190
Myeloid cells staining with myeloperoxidase
Positive with green to dark blue granules
191
Eosinophils staining with myeloperoxidase
Most intense brown-black or green-black granules
192
Neutrophil cytoplasm staining with myeloperoxidase
Blue
193
Basophils lymphocytes and erythroblasts staining with myeloperoxidase
Negative
194
Monocytes staining with myeloperoxidase
Slight peroxidase activity
195
Bone marrow section embedding for Masson’s trichrome
Undecalcified glycol methacrylate embedded sections cut at 4 micrometers
196
Stains in Masson’s trichrome for bone marrow
Osteoid seams red+Mineralized bone blue+Nuclei dark gray
197
Most useful stain for plastic-embedded tissue sections
Toluidine blue
198
Use of reticulin stain
Detection of myelofibrosis+Leukemia+Lymphoma
199
Ancillary procedures in bone marrow analysis
Immunohistochemistry+In-situ hybridization+Cytogenetics+PCR and gene sequencing
200
Types of lymph node biopsies
Fine needle aspiration+Open (excisional) biopsy+Sentinel lymph node biopsy
201
Processing methods for lymph node biopsies
Microscopy on fixed stained tissue+Immunohistochemistry and flow cytometry+Cytogenetic G-banding+FISH+Molecular genetic analysis PCR and sequencing
202
Use of cytogenetic or FISH analysis
Diagnosis of Burkitt’s lymphoma
203
Use of PCR in lymph node biopsies
Detection of specific translocations and T/B cell clonality
