Transcript Acidic (Eosinophilic) and Basic Dyes

To Histo SSN!
Microscopy and Cytology Stains
1.
2.
3.
4.
5.
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 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 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
•
How would you describe
the structure at the
pointer?
a)
b)
c)
d)
Eosinophilic
Basophilic
Negatively charged
Positively charged
Online Reference: Lab #1 Slide #16
Question 1
•
How would you describe
the structure at the
pointer?
a)
b)
c)
d)
Eosinophilic
Basophilic
Negatively charged
Positively charged
Online Reference: Lab #1 Slide #16
Nissl Stain
Lab #1 Slide #1
Lab #1 Slide #2
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
•
What cellular
function is occurring
in this region of this
cell?
a)
b)
c)
d)
Lab #1 Slide #3
rRNA transcription
RNA translation
Protein degradation
DNA degradation
Question 2
•
What cellular
function is occurring
in this region of this
cell?
a)
b)
c)
d)
Lab #1 Slide #3
rRNA transcription
RNA translation
Protein degradation
DNA degradation
Eosin (H&E)
• Pink
• Stains Eosinophilic
structures ex. Proteins,
collagen
Lab #1 Slide #13
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)
• 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
Lab #14 Slide #8
Lab #14 Slide #18
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
•
The pink regions are
eosinophilic due to:
a)
b)
c)
d)
e)
Endosomes
rER
Cytosolic proteins
Golgi
Cilia
Lab 7 #18
Question 3
•
The pink regions are
eosinophilic due to:
a)
b)
c)
d)
e)
Endosomes
rER
Cytosolic proteins
Golgi
Cilia
Lab 7 #18
Question 4
•
The apex of this cell
stains positively
because of
a) Ribosomes
b) Carbohydrates
c) Fats
Lab #14 Slide #18
Question 4
•
The apex of this cell
stains positively
because of
a) Ribosomes
b) Carbohydrates
c) Fats
Lab #14 Slide #18
Question 5
•
The structure at the
pointer
a)
Is composed of
heterochromatin
b) Indicates the cell is
NOT transcriptionally
active
c) Is the primary site of
ribosomal RNA
synthesis
d) Is a Nissl body
Lab 6, #18
Question 5
•
The structure at the
pointer
a)
Is composed of
heterochromatin
b) Indicates the cell is
NOT transcriptionally
active
c) Is the primary site of
ribosomal RNA
synthesis
d) Is a Nissl body
Lab 6, #18
Electron Microscopy
EM 1
Identify structures
EM 1
Identify structures
Nucleus
Outer Nuclear Membrane
Nucleolus
Rough ER
EM1, Q1
Proteins that are made here
may end up at which of
the following sites:
a) Outside the cell
b) Golgi Apparatus
c) Plasma Membrane
d) All of the above
e) None of the above
Lab 1, Slide 4
EM1, Q1
Proteins that are made here
may end up at which of
the following sites:
a) Outside the cell
b) Golgi Apparatus
c) Plasma Membrane
d) All of the above
e) None of the above
Lab 1, Slide 4
EM2
• Active or inactive
cell?
EM2
• Active or inactive
cell?
– ACTIVE
EM2 Q2
Which structure would
appear darker in a
Nissl stain?
B
A
a) A
b) B
c) Neither. They are both
eosinophilic
d) Neither, they do not stain
with Nissl
Lab 1 Sl 7
EM2 Q2
Which structure would
appear darker in a
Nissl stain?
B
A
a) A
b) B
c) Neither. They are both
eosinophilic
d) Neither, they do not stain
with Nissl
Lab 1 Sl 7