Muscle Enzyme Histochemistry

Question 1

Muscle definition

Answer 1

Bundle of tissue that has the ability to contract and produce movement.

Question 2

Types of muscle

Answer 2

Skeletal
Smooth
Cardiac

Question 3

Muscle Breakdown

Answer 3

Fascicle-Fiber-Myofibril-Myofilament-Thick/Thin Filaments

Question 4

Type 1 Fibers

Answer 4

Slow Twitch
Primarily aerobic
Red in poultry
33%
Abundant mitochondria

Question 5

Type 2 Fibers

Answer 5

Fast twitch
Primarily anaerobic
Few mitochondria
Poor blood supply
Rich in glycogen
67%
2A, 2B &2C

Question 6

Three connective tissue covings in the muscle

Answer 6

Collagen
Retic
Elastic Fibers

Question 7

Two types of muscular dystrophy

Answer 7

Duchenne
Becker

Question 8

Symptoms of muscular disease

Answer 8

Weakness
Atrophy
Cramps
Stiffness

Question 9

Microns for muscle sections

Answer 9

8-10µ

Question 10

Best stain for fiber typing

Answer 10

ATPase stain

Question 11

ATPase results

Answer 11

*High pH 9.4
Type 1 light to unstained
Type 2 dark
*Low pH 4.3
Type 1 dark
Type 2A light to unstained
Type 2B intermediate (grey)

Question 12

Which fibers contain the most glycogen? What stain?

Answer 12

Type 2
PAS

Question 13

Muscle biopsy stains

Answer 13

Gomori’s Trichrome
PAS PASD
Oil Red O
NADH
ATPase
Acid Phosphatase
Esterase

Question 14

Stain?
Purpose?

Answer 14

Gomori Trichrome
Demonstration of mitochondrial abnormalities (myopathy, muscle twitches)
Ragged fibers
Nemaline rods
Rimmed vacuoles

Question 15

Trichrome Reagents

Fluffy Gorillas Chase Playful Green Hedgehogs

Answer 15

Fast Green - Connective tissue (green)
Chromotrope - plasma cells
Phosphotung. acid - stains muscle
Glacial acetic acid - sensitizer
Hematoxylin - nuclei (red to purple)
myelinated nerve twigs, nemaline rods & abnormal mitochondria, inetrmyofibrillar sarcoplasm = red

Question 16

Glycocgen storage disease

Answer 16

Indicates increased amount of glycogen

Question 17

Stain?
Purpose?

Answer 17

Demonstrates neutral lipids
Takes form of small droplets w/in myofibrils.
Lipid disorder = large and increased spots
More in Type 1 fibers = more mitochondria where lipids are stored

Question 18

Stain?
Purpose?

Answer 18

NADH
Mitochondrial abnormalities (myopathy)
Indicates architectural changes

Question 19

NADH resuslts

Answer 19

Site of high enzymatic activity in mitochondria = blue to purple
Type 1 fibers = dark
Type 2 = light

Question 20

ATPase pH ranges

Answer 20

Low 4.3 -4.65
High 9.4 - 10.2

Question 21

Stain?
Purpose?

Answer 21

Acid Phosphatase
Locate degeneration
Identifies macrophages related to necrotic fibers
Normal tissue shows minimal to no reaction

Question 22

Stain?
Purpose?

Answer 22

NSE
Enzymatic staining based on denervation
Stains motor end plate connectivity to muscle fibers
Demonstrates inflammatory cells or motor end plates

Question 23

NSE Results

Answer 23

Positive cells = Bright red
Nuclei = Blue
Background = Unstained

Question 24

NSE Reagents

Answer 24

Phosphate Buffer pH 7.2
Pararosaniline stock sltn.
Sodium Nitrite sltn.
a-naphthyl acetate in acetone
HCl
Na Hydroxide
Incubation Sltn.

Question 25

ATPase Reagents

Answer 25

Barbital acetate buffer stock sltn. A " " B " " working sltn. Na Barbital sltn. Incubating sltn. Calcium chloride Cobaltous chloride Ammonium sulfide slt.

Question 26

Acid Phosphatase Reagents Harsh Parachuters Have Salty Nite Vegetables in Substrate Museums

Answer 26

HCl Pararosaniline HCl Na nitrite Veronal acetate buffer Incubating medium Substrate sltn. (no plastic) Methyl green

Question 27

NADH Reagents Salted Porcupines Nap

Answer 27

Saline sltn. Phosphate buffer pH 7.4 Nitro blue tetrazolium (NBT) sltn.