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Welcome. To Histo SSN!. Microscopy and Cytology Stains. Intro to Histo Handout (tips on course) Important terms (Acidophilic, Basic, etc.) Examples of some stains you’ll see Vital stain (of blood smear) EMs Any specific questions you have, please hold to the end. Basophilic.

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slide1

Welcome

To Histo SSN!

microscopy and cytology stains
Microscopy and Cytology Stains
  • Intro to Histo Handout (tips on course)
  • Important terms (Acidophilic, Basic, etc.)
  • Examples of some stains you’ll see
  • Vital stain (of blood smear)
  • EMs

Any specific questions you have,

please hold to the end.

basophilic
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

acidophilic eosinophilic
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

question 1
Question 1
  • How would you describe the structure at the pointer?
    • Eosinophilic
    • Basophilic
    • Negatively charged
    • Positively charged

Online Reference: Lab #1 Slide #16

question 11
Question 1
  • How would you describe the structure at the pointer?
    • Eosinophilic
    • Basophilic
    • Negatively charged
    • Positively charged

Online Reference: Lab #1 Slide #16

nissl stain
Nissl Stain

Lab #1 Slide #1

Lab #1 Slide #2

nissl stain1
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

question 2
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

question 21
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

eosin h e
Eosin (H&E)
  • Pink
  • Stains Eosinophilic structures ex. Proteins, collagen

Lab #1 Slide #13

hemotoxylin h e
Hemotoxylin (H&E)
  • 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

periodic acid schiff hematoxylin pas
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

h e vs pas
H&E vs. PAS

Lab #14 Slide #8

Lab #14 Slide #18

acid fuschin toluidin blue
Acid Fuschin-Toluidin Blue
  • Pink and Blue
    • Pink = acid Fuschin
    • Blue = Toluidin
  • Akin to H&E
  • Note that:
    • Acid dyes stain apical surface
    • Basic dyes stain basal surface

Lab #1 Slide #5

question 3
Question 3
  • The pink regions are eosinophilic due to:
    • Endosomes
    • rER
    • Cytosolic proteins
    • Golgi
    • Cilia

Lab 7 #18

question 31
Question 3
  • The pink regions are eosinophilic due to:
    • Endosomes
    • rER
    • Cytosolic proteins
    • Golgi
    • Cilia

Lab 7 #18

question 4
Question 4
  • The apex of this cell stains positively because of
    • Ribosomes
    • Carbohydrates
    • Fats

Lab #14 Slide #18

question 41
Question 4
  • The apex of this cell stains positively because of
    • Ribosomes
    • Carbohydrates
    • Fats

Lab #14 Slide #18

question 5
Question 5
  • 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

question 51
Question 5
  • 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

em 1 identify structures1
EM 1Identify structures

Nucleus

Outer Nuclear Membrane

Nucleolus

Rough ER

em1 q1
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

em1 q11
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

slide27
EM2
  • Active or inactive cell?
slide28
EM2
  • Active or inactive cell?
    • ACTIVE
em2 q2
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

em2 q21
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