(PALM 305) Exam 2 objectives Flashcards
1
Q
GaGs
A
all acidic in connective tissue
2
Q
Glycoproteins/mucins/muosubstances
A
epithelial secreting mucin cells
3
Q
PAS Purpose
A
insoluble carbohydrates
4
Q
PAS results
A
BM, glycogen, fungal walls, neutral and acidic proteins: Magenta
Background: Blue/purple
5
Q
A
PAS ID
6
Q
A
PAS mucin ID
7
Q
A
PAS Fungus ID
8
Q
How do you test the quality of a schiff reagent
A
place a few drops in a beaker of 37-40% formaldehyde, if the solution is reddish purple the reagent is good
9
Q
Most common cause from weak staining with PAS?
A
insufficient water rinsing
10
Q
How do you test the quality of a schiff reagent
A
place a few drops in a beaker of 37-40% formaldehyde, if the solution is reddish purple the reagent is good
11
Q
Most common cause from weak staining with PAS?
A
insufficient water rinsing
12
Q
PAS with digestion purpose
A
to demonstrate glycogen in tissue
13
Q
Quality control of PAS with digestion
A
liver, cervix, vaginia
14
Q
PAS with digestion results
A
without digestion: glycogen and all carbohydrates are dark pink.red
With digestion: no glycogen and all other carbohydrates are pink
Background: purple
15
Q
PAS with digestion troubleshooting
Why would digestion fail?
A
-insufficient digestion time
-overheated or insufficiently heated malt disease
-Tissue fixed in picric acid containing fixatives
16
Q
A
PAS with digestion vs without
17
Q
A
PAS w digestion
18
Q
A
PAS w/o digestion
19
Q
This stain stains only glycogen
A
Best Carmine
20
Q
A
Best Carmine
21
Q
Mucicarmine purpose
A
Acidic glycoproteins , fungus
22
Q
What is weigert solution A and B made up of in mucicarmine?
A
Solution A: alcoholic hematoxylin
Solution B:Mordant ferric chloride
23
Q
What is the quality control for Mucicarmine?
A
GI specimen, colon preferred, tissue has to be not autolyzed
24
Q
Mucicarmine results
A
Acidic glycoproteins- pink/magenta
Nuclei- blue/black
BAckground- yellow/green
25
Mucicarmine ID
26
Mucicarmine demonstrates
Acidic Glycoproteins
27
~~ weak or pale staining with mucicarmine
if weigert's hematoxylin is used
28
~~ mucins are obscured in mucicarmine staining
over counterstaining with, using Gill
29
~~ weak mucin staining mucicarmine
not left in solution long enough, over rinsing
30
Absent mucin staining mucicarmine
Autolyzed tissue, enzymes eat the mucins
31
Alcian blue 2.5 purpose
demonstrates GaGs and acidic glycoproteins
32
Why must you rinse with acetic acid before and after alcian blue staining?
to maintain pH and prevent nonspecific staining
33
What is the recommended pH for alcian blue?
2.5, the dye is selective
34
quality control for alcian blue 2.5
GI specimens (olon), appendix
35
Alcian blue 2.5 results
Acidic glycoproteins: blue (intense)
Gags: blue, less intense
Background/nuclei: pink/red
36
Alcian Blue 2.5
37
What could cause excessive background staining in alcian blue 2.5
A pH over 2.5
38
Alcian Blue 2.5 (Carboxylated and Sulfates polysaccharides)
39
Alcian Blue 1.0 (Sulfated polysaccharides)
40
What does alcian blue 1.0 demonstrate
sulfomucin (sulfated and carboxylated, GaGs)
41
Results of Alcian blue 1.0
Sulfated GaGs and sulfomucins: pale blue
Background/nuclei: pink from counterstain
42
Quality control Alcian blue 1.0
Large intestine, appendix
43
Alcian blue 2.5 with Hyaluronidase purpose
digests GaGs
44
Principle and reagents for alcian blue 2.5 with hyaluronidase
duplicate slides are required, one treated with hyaluronidase and one without.
-both sides are then stained with the same alcian blue 2.5 procedure
45
what is the quality control for alcian blue 2.5 with hyaluronidase?
2 slides of umbilical cord
-small intestine and appendix used as second control
46
Results of Alcian blue 2.5 with hyaluronidase
without digestion (GaGs and Glycoproteins)- blue
With digestion- no GaGs glycoproteins still blue
Background- pink/red
47
Alcian blue 2.5 with hyaluronidase procedural notes
hyaluronidase digestion can occur with colloidal iron method
rinse well after NFR counterstain or cloudiness will occur
48
Alcian Blue 2.5 without Hyaluronidase
49
Alcian Blue 2.5 with Hyaluronidase
50
Alcian blue PAS purpose
demonstrates neutral and acidic glycoproteins
51
diagnosis barrett's esophagus
Alcian blue PAS quality control
duodenum, endocervix, ectocervix
52
Alcian blue PAS results
GaGs and acidic glycoproteins- Blue
Neutral mucosubstances and glycogen: pink
Mixture of neutral and acidic mucosubstances: purple
Nuclei: blue/purple
53
Alcian Blue PAS
54
Colloidal iron purpose
demonstrates acidic glycoproteins
55
what do ferric ions react with and what do they form in colloidal iron staining?
potassium ferrocyanide, Prussian Blue
56
Colloidal iron is _____than alcian blue but more _______
specific, sensitive
57
Results of colloidal iron
GaGs and glycoproteins: blue
Counterstain: nuclear fast red
acid fuchsin: collagen pink/red
picric acid: muscles and cytoplasm yellow
58
what can also stain positive in colloidal iron and how can you prevent that?
Hemosiderin, prevent by adding potassium ferrocyanide
59
Colloidal iron
60
Colloidal iron Van glenson counterstain
61
Purpose of high iron diamine
demonstrates sulfomucins
-when combined with alcian blue, it can differentiate between sulfomucins and sialomucins
62
Results of High iron Diamine
sulfomucin: black/brown
Sialomucins and Gags: Blue
63
High Iron Diamine
64
Troubleshooting with High iron Diamine
prolonged time in alcian blue may result in nuclear staining
rinse well after NFR or cloudiness will occur
65
Amyloid composition
parallel beta pleated sheets
abnormal, large, extracellular protein
66
Appearance of amyloid in H&E and congo red
H&E- eosinophilic/pink
Congo red: pink/red
67
Primary amyloidosis
igG LIGHT Chains deposit
plasma cell disorder
68
Secondary Amyloidosis
associated with inflammatory disease
69
Purpose of alkaline red congo
demonstrating amyloid
70
What is the primary staining mechanism on alkaline red congo
Van Der Waals
71
What is the primary staining mechanism on alkaline red congo
Van Der Waals
72
what thickness is amyloid stains cut at?
8-10 microns
73
Results of alkaline congo red stain
Amyloid: pink/red, Apple green with polarized microscope
Background: blue
74
Congo Red
75
Congo Red w polarized microscope
76
Crystal/methyl violet
77
Birefringence is most observed using which microscope
polarized microscope
78
Crystal/methyl violet purpose
Amyloid demonstration, rarely used today
79
Results of crystal/methyl violet
Amyloid: intense violet
Background: less intense violet
80
Why do you have to use alternative coverslipping methods for crystal/methyl violet and thioflavin stains?
you have to avoid alcohols
81
Thioflavin purpose
demonstrates amyloids
82
Thioflavin principle and reagents
mayer hematoxylin: counterstain that prevents fluorescence of nuclei
Thioflavin T: fluorescent dyes that react with amyloid
83
Results of thioflavin staining
Amyloid: fluorescent yellow/green
Nuclei/background: blue
84
Thioflavin ID
85
Does polarized or fluorescence allow you to see tissue detail in the background?
Polarized allows you to see tissue detail
fluorescent has a pitch black background
86
Fuelgen no counterstain
87
Fuelgen w counterstain
88
What does the feulgen reaction demonstrate
DNA only
89
in the feulgen reaction schiff reagent reacts with?
aldehydes of DNA, after the rinse step magenta color appears
90
what is the most preferred fixative in the feulgen stain
Alcohol
91
Results of Feulgen stain
DNA- Pink
Cytoplasm/background- unstained, or light green of a counterstain is used
92
What is the purpose of the methyl green Pyronin reaction
Demonstrates RNA and DNA
93
Methyl green reacts with what polymer
DNA
94
Pyronin Y reacts with what polymer
RNA
95
Results of Methyl- green Pyroin Y staining
DNA- Green
RNA- Pink
Plasma Cells/cytoplasm: pink/red
96
Methyl Green Pyronin Y
97
Methyl Green Pyronin Y
98
Neutral/romanowsky
combination of cationic and anionic dyes
99
Polychromatic dyes
homologues resulting in different color results
100
Polychroming process
process where a dye forms another dye, initiated and controlled by pH
101
what is the purpose of the Giemsa stain
to demonstrate hematopoietic cells
also microorganisms
102
Giemsa microns
3-4 microns
103
Quality control for giemsa staining
Bone marrow
Spleen
104
Results of Giemsa staining
Blue:
Nuclei + Bacteria
Violet/Purple:
Basophils + Neutrophils + Mast cells
Pink:
Eosinophils
105
Giemsa in smear
106
Giemsa in Bone marrow
107
Giemsa in Bone Marrow
108
The first evidence of under processing will be present in connective tissue because they are
Hydrophilic
109
What are examples of GaGs
chondroitin and hyaluronic acid
110
What is the pathological significance of demonstrating GaGs
Sarcomas
111
What is the pathological significance of demonstrating glycoproteins
Carcinomas
112
Diastase is used in the PAS procedure to digest what
glycogen
113
Fungus will demonstrate with what stains?
Mucicarmine
Alcian Blue
PAS
114
That water rinse in the PAS procedure is critical in order to restore what
the chromophore
115
Which fixative can render glycogen resistant to digestion
Bouins
116
Best carmine will demonstrate
glycogen
117
Mucicarmine demonstrates what
acidic glycoproteins
118
Alcian blue 2.5 will demonstrate what?
Sulfated GaGs and glycoproteins
119
High iron diamine differentiates what?
sialomucins and sulfomucins
120
Prolonged fixation in formalin will cause what effect on the congo red staining?
false negative
121
A polychromatic dye that demonstrates amyloid is?
Crystal violet
122
in the congo red method, which solution disrupts hydrogen bonding of amyloid and contributes to the increased specificity?
alkaline salt solution
123
The schiff reagent will demonstrate all of the following except?
Nuclei
124
Polysaccharides within connective tissue are referred to as?
GaGs
125
Which of the following fixatives should be avoided for the PAS procedure?
Glutaraldehyde
126
Glycogen is a component of which group of carbohydrates?
neutral polysaccharides
127
Epithelial polysaccharides commonly located in mucous producing cells are referred to as?
glycoproteins
128
Which of the following could be a cause of excessive background staining in Alcian blue pH 2.5?
a pH over 2.5
129
Hyaluronidase digestion will remove
GaGs
130
Alcian blue pH 2.5 demonstrates
all acidic polysaccharides
131
The mucicarmine procedure demonstrates
acidic glycoproteins
132
A pH of 1.0 in the Alcian blue procedure is important to exclusively demonstrate?
carboxylated and sulfated GaGs and sulfomicuns
133
Amyloid is a?
Abnormal protein
134
the primary staining mechanism for the congo red method is
Van Der Waals
135
Which of the following is a fluorescent method for the demonstration of amyloid?
Thioflavin T
136
An appropriate control tissue for the colloidal iron method would be
colon
137
The insoluble blue color formed in the colloidal iron method is referred to as
purassin blue
