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Musculoskeletal System > 7.Histology of MSK system > Flashcards

7.Histology of MSK system Flashcards

1
Q

what is HISTOLOGY

A

EXAMINING TISSUES/CELLS using a MICROSCOPE

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2
Q

HISTOLOGY needs PREPARATION of the TISSUE. 4 steps:

A
  • FIXATION
  • EMBEDDING
  • SECTIONING
  • STAINING
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3
Q

Why do we need FIXATION (4)

A
  • To PRESERVE tissue
  • To PROTECT and FIX STRUCTURES
  • To make INTRACELLULAR STRUCTURES ACCESSIBLE to reagents
    (Immuno/histochemistry)
  • To STOP a PROCESS at a CERTAIN POINT in time
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4
Q

2 COMMON FIXATION METHODS

A
  • CHEMICAL FIXATIVES such as formaldehyde, EtOH
    (chemicals may alter tissue)
  • FREEZING
    (structures less well-defined)
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5
Q

why do we need EMBEDDING of tissues (2)

A
  • to SUPPORT the sample DURING SECTIONING
  • to PRESERVE FRAGILE 3D STRUCTURES
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6
Q

3 COMMON EMBEDDING MATERIALS

A
  • WAX (most common)
  • SUCROSE solutions for cry-sectioning (frozen samples)
  • Hard PLASTICS for HARD TISSUES eg bone, teeth
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7
Q

WHY do we SECTION tissues

A

to produce samples that are THIN enough
for the Microscope LIGHT to SHINE THROUGH

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8
Q

most common INSTRUMENT for SECTIONING is called

A

ROTARY MICROTOME

(can change thickness)

TYPES of KNIVES used:
steel (MOST)
diamaond,
glass

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9
Q

why is STAINING important

A
  • normal tissues show very little contrast
    staining needed to INCREASE CONTRAST

& DIFFERENTIATE between STRUCTURES with SIMILAR CONTRASTS

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10
Q

example of a general STAIN used

A

HAEMATOXYLIN/EOSIN STAIN (H&E)

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11
Q

H&E STAINS NUCLEI which colour

A

PURPLE

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12
Q

2 STAINS used for ELASTIC FIBRES (eg Dermis)

A
  • MASSON’S TRICHROME (PURPLE)
  • VERHOEFF’S STAIN
    (PURPLE elastic fibres but other structures RED)
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13
Q

stain for Reticular Fibres (in bone marrow)

A

Collagen Type III STAIN

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14
Q

HISTOLOGY of SKELETAL MUSCLE

(nucleus, striations, intercalated borders, branching, size of fibre)

A

MULTI-NUCLEATED
nucleus is PERIPHERAL not central

STRIATED

LARGE Fibre size

no intercalated borders
no branching

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15
Q

HISTOLOGY of CARDIAC MUSCLE

(nucleus, striations, intercalated borders, branching, size of fibre)

A

ONE NUCLEUS
CENTRAL

STRIATED

INTERCALATED BORDERS (intercalated DISCS where attach to each other)

BRANCHING

LARGE SIZE

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16
Q

HISTOLOGY of SMOOTH MUSCLE (eg small intestine)

(nucleus, striations, intercalated borders, branching, size of fibre)

A

ONE NUCLEUS,
CENTRAL

SMALL SIZE

2 LAYERS - PERPENDICULAR

not striated,
no borders,
no branching

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17
Q

how are SMOOTH MUSCLE LAYERS in SMALL INTESTINE

(stain H&E)

A

2 LAYERS

fibres in each layer are PERPENDICULAR to each other

INNER LAYER: LONGITUDINAL

OUTER LAYER: CROSS SECTIONAL

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18
Q

STAINS used for SKELETAL MUSCLE (3 examples)

A

H&E (RED, PURPLE)

STAIN FOR ATPase ACTIVITY (shows type 1 and type 2) (brown)

SIRIUS RED STAIN - STAINS COLLAGEN in the ECM
- RED (rest is YELLOW)

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19
Q

what does the ATPase STAIN show in skeletal muscle

A

TYPE 1 - SLOW TWITCH & TYPE 2 - FAST TWITCH muscle

ATPase pH 9.4
(Colours switched at pH 4.3)

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20
Q

how is the NUCLEUS in SKELETAL MUSCLE

A

MULTI-NUCLEATED
PERIPHERAL

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21
Q

how is the NUCLEUS in SKELETAL MUSCLE

A

MULTI-NUCLEATED
PERIPHERAL

22
Q

Which types of MUSCLE has 1 NUCLEUS per cell/fibre

A

CARDIAC
& SMOOTH

23
Q

which muscle type has BRANCHING

A

CARDIAC

24
Q

which muscle types have STRIATIONS

A

SKELETAL
CARDIAC

25
Q

2 LAYERS OF SMOOTH MUSCLE:
(small intestine)

A

Inner layer: LONGITUDINAL
Outer layer: CROSS SECTIONAL

26
Q

which muscle type has INTERCALATED BORDERS

A

CARDIAC

27
Q

which muscle type has SMALL fibre size

A

SMOOTH

28
Q

how are COLLAGEN FIBRES arranged in TENDONS & LIGAMENTS

A

LONGITUDINALLY

(tendon: parallel, ligament: cross-connecting)

29
Q

COLLAGEN types in TENDONS & LIGAMENTS

A

TYPE 1

some TYPE 3 INCREASES AFTER INJURY

30
Q

composition of ELASTIC CARTILAGE

A

HIGH CELL DENSITY
CHONDROCYTES (rounded appearance) in lacunae
collagaen fibres
elastic fibres

31
Q

which STAIN allows ELASTIC FIBRES to be VISIBLE in ELASTIC CARTILAGE (opposed to H&E)

A

MASSON’S TRICHROME

32
Q

what colour do ELASTIC FIBRES appear in elastic cartilage using MASSON’S TRICHROME STAIN

A

DARK PURPLE

33
Q

CHONDROCYTES APPEARANCE:

A

ROUND

34
Q

how is CELL DENSITY in FIBROCARTILAGE as opposed to in elastic cartilage

A

INTERMEDIATE CELL DENSITY
- NOT as closely packed as in elastic cartilage

35
Q

how is CELL DENSITY in HYALINE CARTILAGE as opposed to elastic cartilage and fibrocartilage

A

LOWER CELL DENSITY
- space around cells

36
Q

COLLAGEN in FIBROCARTILAGE

A

TYPE 1 & TYPE 2

37
Q

what ‘mark’ do you find in ARTICULAR CARTILAGE

A

TIDE MARK
- BOUNDARY for CALCIFIED CARTILAGE

38
Q

besides H&E, what other STAINS can be used for ARTICULAR CARTILAGE
(what colours)

A

SAFRANIN O
& FAST GREEN
(with Haematoxylin counterstain)

  • stains GLYCOPROTEINS: ORANGE
  • TYPE 1 COLLAGEN: GREEN
  • cells: PURPLE
39
Q

why is HISTOLOGY of bone DIFFICULT

A

bone is MINERALISED

40
Q

2 options for HISTOLOGY of bone

A
  1. DEMINERALISE -> STANDARD WAX HISTOLOGY
  2. DON’T demineralise -> EMBED in HARD RESIN
41
Q

ADVANTAGES and DISADVANTAGES in DEMINERALISING bone for HISTOLOGY (& using standard wax histology)

A

adv: ALL NORMAL STAINS AVAILABLE

disadv: INFORMATION on the bone MINERALISATION is LOST

42
Q

ADVANTAGES and DISADVANTAGES in NOT DEMINERALISING & EMBEDDING IN HARD RESIN for HISTOLOGY

A

adv: MINERALISATION INFORMATION RETAINED

disadv:
-SPECIAL SECTIONING EQUIPMENT required
- NOT ALL stains work

43
Q

how is CORTICAL BONE

A

DENSE,
MOSTLY mineralised bone matrix

can see OSTEOCYTES (mechanoreceptors)
OSTEONS (contain blood vessels)

Osteoblasts on edge

44
Q

how is TRABECULAR BONE

A

OPEN STRUCTURE,
ABUNDANT BONE MARROW SPACE

Adipocytes, Osteoblasts on edge,

45
Q

which bone cells are RARE to find and what are they like

A

OSTEOCLASTS

  • LARGE
  • MULTI-NUCLEATED
46
Q

what can you EMBED UNDEMINERALISED (mineralised) bone in

A

Methyl-Methacrylate

47
Q

STAIN you can use for MINERALISED / UNDEMINERALISED bone and what does it allow you to see

A

GOLDNERS TRICHROME STAIN

  • can identify mineralised bone (GREEN)
  • can see NEW BONE TISSUE that has NOT YET BEEN MINERALISED (ORANGE)
48
Q

VAN KOSSA STAIN
stains mineralised bone:
stains osteoid (new bone, not yet mineralised):

A

mineralised: BLACK

new bone: PINK

49
Q

Calcein Double Labelling allows you to see..

(in mineralised)

A

PROGESS of BONE GROWTH
(rate)

50
Q

STAIN for OSTEOCLASTS

A

TRAP stain
(RED)
stain TRAP enzymes in osteoclasts

(mineralised bone BLUE)

Musculoskeletal System (18 decks)

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