Acidic (Eosinophilic) and Basic Dyes

1
Welcome
To Histo SSN!
2
Microscopy and Cytology Stains

• Intro to Histo Handout (tips on course)
• Important terms (Acidophilic, Basic, etc.)
• Examples of some stains youll see
• Vital stain (of blood smear)
• EMs
• Any specific questions you have,
• please hold to the end.

3
Basophilic

• Basophilic structures are stained by basic dyes
• Basic dyes are positive
• Basophilic structures are negative (ex. DNA, RNA,
  ribosomes, RER)
• Mnemonic Basophilic Blue
• Online Reference Lab 1 Slide 14

4
Acidophilic (Eosinophilic)

• Acidophilic structures are stained by acid dyes
• Acid dyes are negative
• Acidophilic structures are positive (ex.
  Proteins, collagen, cytoplasm)
• Eosinophilic Pink
• Online Reference Lab 1 Slide 14

5
Question 1

• How would you describe the structure at the
  pointer?
• Eosinophilic
• Basophilic
• Negatively charged
• Positively charged
• Online Reference Lab 1 Slide 16

6
Question 1

• How would you describe the structure at the
  pointer?
• Eosinophilic
• Basophilic
• Negatively charged
• Positively charged
• Online Reference Lab 1 Slide 16

7
Nissl Stain
Lab 1 Slide 1
Lab 1 Slide 2
8
Nissl Stain

• Blackish blue
• Stains negatively charged structures
• Nissl substance (bodies) stacks of RER and free
  polyribosomes
• Nucleolus site of rRNA synthesis
• Lab 1 Slide 3

9
Question 2

• What cellular function is occurring in this
  region of this cell?
• rRNA transcription
• RNA translation
• Protein degradation
• DNA degradation
• Lab 1 Slide 3

10
Question 2

• What cellular function is occurring in this
  region of this cell?
• rRNA transcription
• RNA translation
• Protein degradation
• DNA degradation
• Lab 1 Slide 3

11
Eosin (HE)

• Pink
• Stains Eosinophilic structures ex. Proteins,
  collagen
• Lab 1 Slide 13

12
Hemotoxylin (HE)

• Blue, purple or blackish
• Stains Basophilic structures ex. DNA, ribosomes,
  RNA
• Euchromatin is DNA in USE. It is spread out,
  diffuse, and less stained.
• Heterochromatin is condensed DNA, and stains dark
  blue.
• Lab 1 Slide 13

13
Periodic Acid Schiff Hematoxylin (PAS)

• Pink, Magenta
• Stains carbohydrates and carb. rich
  macromolecules ex. Glycogen, mucin, basement
  membrane, etc.
• If you see PAS, think CARBOHYDRATES.
• Lab 1 Slide 10

14
HE vs. PAS
Lab 14 Slide 8
Lab 14 Slide 18
15
Acid Fuschin-Toluidin Blue

• Pink and Blue
• Pink acid Fuschin
• Blue Toluidin
• Akin to HE
• Note that
• Acid dyes stain apical surface
• Basic dyes stain basal surface
• Lab 1 Slide 5

16
Vital Stains(of Blood Smears)
17
Neutral Red

• Red when protonated
• Colorless and permeable when unprotonated
• Accumulates in Lysosomes, which are acidic ?
  protonation
• Lab 1 Slide 19

18
Janus Green

• Green when oxidized
• Colorless and permeable before oxidation
• Accumulates in mitochondria, which contain
  oxidative agents ? oxidation
• Lab 1 Slide 19

19
Question 3

• Why are the organelles seen here stained red?
• There is an oxidative environment
• There is an acidic environment
• Proteins are being degraded
• RNA translation is occurring
• They are embarrassed
• Lab 1 Slide 18

20
Question 3

• Why are the organelles seen here stained red?
• There is an oxidative environment
• There is an acidic environment
• Proteins are being degraded
• RNA translation is occurring
• They are embarrassed
• Lab 1 Slide 18

21
Question 4

• The pink regions are eosinophilic due to
• Endosomes
• rER
• Cytosolic proteins
• Golgi
• Cilia
• Lab 7 18

22
Question 4

• The pink regions are eosinophilic due to
• Endosomes
• rER
• Cytosolic proteins
• Golgi
• Cilia
• Lab 7 18

23
Question 5

• The apex of this cell stains positively because
  of
• Ribosomes
• Carbohydrates
• Fats
• Lab 14 Slide 18

24
Question 5

• The apex of this cell stains positively because
  of
• Ribosomes
• Carbohydrates
• Fats
• Lab 14 Slide 18

25
Question 6

• The structure at the pointer
• Is composed of heterochromatin
• Indicates the cell is NOT transcriptionally
  active
• Is the primary site of ribosomal RNA synthesis
• Is a Nissl body
• Lab 6, 18

26
Question 6

• The structure at the pointer
• Is composed of heterochromatin
• Indicates the cell is NOT transcriptionally
  active
• Is the primary site of ribosomal RNA synthesis
• Is a Nissl body
• Lab 6, 18

27
Electron Microscopy
28
EM 1Identify structures
29
EM 1Identify structures
Nucleus
Outer Nuclear Membrane
Nucleolus
Rough ER
30
EM1, Q1

• Proteins that are made here may end up at which
  of the following sites
• Outside the cell
• Golgi Apparatus
• Plasma Membrane
• All of the above
• None of the above
• Lab 1, Slide 4

31
EM1, Q1

• Proteins that are made here may end up at which
  of the following sites
• Outside the cell
• Golgi Apparatus
• Plasma Membrane
• All of the above
• None of the above
• Lab 1, Slide 4

32
EM2

• Active or inactive cell?

33
EM2

• Active or inactive cell?
• ACTIVE

34
EM2 Q2

• Which structure would appear darker in a Nissl
  stain?
• A
• B
• Neither. They are both eosinophilic
• Neither, they do not stain with Nissl
• Lab 1 Sl 7

B
A
35
EM2 Q2

• Which structure would appear darker in a Nissl
  stain?
• A
• B
• Neither. They are both eosinophilic
• Neither, they do not stain with Nissl
• Lab 1 Sl 7

B
A
36
EM3

• Identify the Structures Functions . . .
• Lab 1 Sl 11

37
EM3 Q3

• The structure at the pointer would be
  demonstrated with which of the following
• HE
• PAS
• Janus Green
• Nissl
• Lab 1 Sl 11

38
EM3 Q3

• The structure at the pointer would be
  demonstrated with which of the following
• HE
• PAS
• Janus Green
• Nissl
• Lab 1 Sl 11