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Histology HTL Exam- BOC Study Guide > Chapter 6: Nuclear and Cytoplasmic Staining > Flashcards

Chapter 6: Nuclear and Cytoplasmic Staining Flashcards

1
Q

What type of tissue is Harris hematoxylin used to stain?

A

Nuclei.

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2
Q

How do you “ripen” hematoxylin?

A

You expose it to oxygen.

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3
Q

What is the active staining molecule in ripened hematoxylin called?

A

Hematein.

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4
Q

What is the most important step in regressive hematoxylin staining?

A

The differentiation in acid alcohol because regressive hematoxylin staining is overstaining then differentiating.

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5
Q

Hematein is formed in Mayer hematoxylin solution by the addition of:

A

Sodium Iodate in order to “ripen” (oxidate) the hematoxylin.

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6
Q

How are mordants used in stains?

A

Oxidize the staining solution.

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7
Q

During H&E staining, if ammonia is incompletely removed by washing, the result may be:

A

Poor eosin staining.

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8
Q

The combination of a dye and a mordant is called a:

A

Lake.

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9
Q

Mercuric oxide was used in the original formula for Harris hematoxylin to:

A

Form hematein, but it was extremely toxic so it was replaced.

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10
Q

Which dye can demonstrate DNA?

A

The Feulgen reaction as well as methyl green.

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11
Q

T/F: Weigert is considered an Iron hematoxylin.

A

True.

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12
Q

What kind of hematoxylin is not readily decolorized with acidic solutions?

A

Iron hematoxylins, such as Weigert.

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13
Q

Why are natural resins hardly used for mounting sections?

A

Natural resins such as Canada balsam and gum dammar are slightly acidic and cause tissue fading after several years.

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14
Q

What kind of stain requires the use of an aqueous mounting media?

A

Oil red O so it doesn’t disrupt the lipid laden macrophages.

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15
Q

What type of metal salt serves as the mordant in Weigert hematoxylin?

A

Iron

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16
Q

Why is acetic acid added to Harris hematoxylin?

A

To make nuclear staining more specific.

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17
Q

What is a dye that can be substituted for hematoxylin in routine staining?

A

Celestine blue can be substituted for hematoxylin with identical results.

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18
Q

What is a good example of a polychrome stain?

A

Giemsa.

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19
Q

What is a polychrome stain?

A

A polychrome stain is a compound dye or dye mixture containing components of different colors.

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20
Q

What cellular structure is stained rose by the methyl green-pyronin (MGP) technique?

A

RNA.

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21
Q

The chemical group in dyes that confers the property of color is called a:

A

Chromophores.

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22
Q

What is an auxochrome?

A

Ionizing groups that confer to the tissue.

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23
Q

What is formed when hematoxylin is subjected to the action of sodium iodate?

A

Hematein.

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24
Q

What is heterochromatin stained by?

A

Heterochromatin is the substance in the nucleus that is stained by hematoxylin.

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25
Q

What stain is a good example of dye absorption?

A

The oil red O because all of the dye is soaked up by the lipids.

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26
Q

If placed in a solution with a pH below the IEP, cytoplasmic proteins will be:

A

At a pH of below 6.0, the cytoplasmic proteins will develop a dominance of positive charges and will attract acid dyes.

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27
Q

How do you differentiate the hematoxylin in H&E stains?

A

A weak acid (acid alcohol).

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28
Q

A tissue component that takes up cationic (positively charged) dye is said to be:

A

Basophilic.

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29
Q

What dye category does eosin fall into?

A

An anionic (acidic) dye.

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30
Q

For best cytoplasmic staining, the pH of eosin should be in between:

A

4.6 and 5.0.

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31
Q

What is the function of ethylene glycol in hematoxylin solutions?

A

It prevents a surface sheen.

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32
Q

What hematoxylin stain will stain the mucin in goblet cells?

A

Gill, because it does not have an excess of aluminum ions to out compete the hematoxylin.

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33
Q

What cellular structures does the Feulgen reaction demonstrate?

A

DNA only.

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34
Q

The Feulgen reaction is unsatisfactory on tissue fixed in:

A

Bouin solution, because it hydrolyzes the nuclei excessively.

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35
Q

Romanowsky type stains are a combination of:

A

Acid and base dyes.

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36
Q

Nuclear staining is made more selective by adding what to the hematoxylin?

A

Acetic acid.

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37
Q

Hematoxylin is converted to a dye lake in the Mayer formula by adding what to the solution?

A

Either ammonium or potassium aluminum sulfate.

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38
Q

A large quantity of Delafield hematoxylin stock solution must be maintained because:

A

Delafield hematoxylin is oxidized by air and light so a large quantity has to be maintained because of how slow the oxidation process is.

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39
Q

To link hematoxylin to tissue DNA, what must be added?

A

A mordant.

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40
Q

When does hematoxylin officially become a dye?

A

Is oxidized.

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41
Q

Romanowsky type stains are preferred for the demonstration of:

A

Leukocytes.

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42
Q

For the most transparency and clarity when viewing well stained microscopic section, the refractive index of the mounting medium should be:

A

Near that of the tissue so the tissue will become more transparent.

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43
Q

Synthetic resins are preferred over natural resins because:

A

Synthetic resins harden more quickly, don’t cause fading of the tissues and do not yellow over time.

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44
Q

What is the approximate refractive index of aqueous mounting media?

A

Aqueous mounting media has a refractive index that is significantly below the tissue.

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45
Q

How can a decrease in section transparency be caused?

A

The transparency of the section decreases if the mounting media becomes too thick.

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46
Q

Fixation with NBF will cause the cytoplasm to:

A

Take up more hematoxylin because the formalin binds with the -NH2 groups which leaves less of these groups to bond with eosin.

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47
Q

If the pH is between 4.6 and 5.0, what chemical groups will the eosin bind to?

A

-NH3.

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48
Q

What can be added to aqueous mounting medium to prevent bleeding of aniline dyes into the surrounding medium?

A

Cane sugar.

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49
Q

A Giemsa is most satisfactory if the pH is between:

A

6.4 and 6.9.

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50
Q

What hematoxylin solution must be used within a few days after preparation?

A

Weigert.

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51
Q

What organelle causes the cytoplasm to show increased basophilia?

A

The rough endoplasmic reticulum because of the large amount of ribosomes in it.

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52
Q

T/F: Basic dyes have a negative charge.

A

False, they’re cationic or positively charged dyes.

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53
Q

T/F: Oxidizers are sometimes used for differentiation.

A

True.

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54
Q

T/F: Eosin is differentiated by the dehydrating alcohols.

A

True, especially the lower ones like 70%.

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55
Q

T/F: An increase or decrease in the pH of the staining solutions can alter staining by changing tissue/dye charges.

A

True.

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56
Q

T/F: Staining can be influenced by the fixative used.

A

True.

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57
Q

T/F: Gill hematoxylin is an iron hematoxylin.

A

False, Gill is an aluminum hematoxylin.

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58
Q

T/F: Ferric chloride is both a mordant and an oxidizer.

A

True.

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59
Q

T/F: An increase in temperature usually increases the rate of staining.

A

True.

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60
Q

T/F: Toluene Blue can stain frozen sections.

A

True.

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61
Q

T/F: Resinous mounting media can have an index of refraction much lower than that of the tissue.

A

False, the refraction index must be very close to that of the tissue.

62
Q

T/F: Resinous mounting media are usually dissolved in toluene, xylene or a xylene substitute.

A

True.

63
Q

T/F: “The more the better” is a good rule of thumb for using mounting medium.

A

False, too much mounting medium can blur the tissue.

64
Q

T/F: The nucleolus of plasma cells is stained green with the methyl green-pyronin technique.

A

False, the RNA is stained by this technique.

65
Q

T/F: Bouin solution is a good choice for tissue to be stained with the Feulgen technique.

A

False, it would excessively hydrolyze the nuclei.

66
Q

Microscopic evaluation of an H&E stained section shows nuclei with well defined chromatin patterns, crisp nuclear membranes and very pale pink eosin staining of RBCs and cytoplasms. What happened?

A

Incomplete removal of the bluing will cause inadequate eosin staining, there is a pH issue with the eosin.

67
Q

An H&E stained section shows reddish brown stained nuclei, pink cytoplasm and bright rose-red RBCs. This would indicate:

A

Overripened hematoxylin will cause this.

68
Q

An H&E stained section shows very uneven staining, with some areas of the slide well stained with other areas unstained. What happened?

A

The paraffin was not completely removed during staining.

69
Q

While staining a rack of sections, it is noted that the water following the rehydrating alcohols is very cloudy. What’s wrong?

A

The xylene has been carried over into the water, the alcohols need to be changed.

70
Q

H&E stained sections reveal brown pigmentlike stippling and rare black nuclei. What happened?

A

The mounting medium has dried before coverslipping.

71
Q

H&E sections of liver show very dark nuclei and some blue staining of the cytoplasm. What happened?

A

There was inadequate hematoxylin differentiation.

72
Q

H&E stained sections show that RBCs, collagen and muscle are all stained the same color. What happened?

A

Poor differentiation of eosin, there should be three different shades of pink.

73
Q

A blue-black precipitate is seen on H&E stained sections. This could be prevented in the future by:

A

Filtering the hematoxylin to avoid these crystals that are formed during the ripening process.

74
Q

Water bubbles are seen during H&E evaluation. This can be prevented in the future by:

A

Making sure that the dehydration process has been completed.

75
Q

Microscopic evaluation of a kidney section is very difficult. Some areas are difficult to focus on while others are perfectly clear. This is most likely because:

A

Mounting medium is on top of the coverglass.

76
Q

H&E stained sections show hazy blue nuclei, but recuts from tissue processed a week earlier show excellent nuclear staining. One possible cause of this problem is:

A

The use of too much heat during processing.

77
Q

A Feulgen stain is requested on a section of Bouin fixed lymph node. How can you accomplish this?

A

Tissue fixed in another solution should be requested as the nuclei has already been hydrolyzed excessively by the Bouin solution.

78
Q

Sections for specials have accidentally already been stained with hematoxylin. How can you remove the hematoxylin?

A

Acid alcohol.

79
Q

The last dehydrating 100% alcohol in the stainer is very pink. This would indicate:

A

Since the eosin is usually differentiated by the lower alcohol concentrations, this would indicate that this absolute alcohol has been contaminated with water.

80
Q

How often should the solutions be changed on the stainer?

A

Solutions should be rotated based on the number of slides that will be going through the stainer.

81
Q

Giemsa staining is very poor on a bone marrow specimen. This problem can be corrected in the future by:

A

The pH of the staining solution needs to be changed, that’s probably why we make our Giemsa solution from scratch every time.

82
Q

Microscopic review of an H&E shows an artefact known as “cornflaking”. What happened?

A

The slide has been dried prior to mounting.

83
Q

What color are mycobacteria stained during the Fite procedure?

A

Red.

84
Q

What stain will bind to acid mucosubstances, which can be then demonstrated during the Prussian blue reaction?

A

Colliodal iron.

85
Q

Duplicate sections are stained with PAS, one with diastase and one without digestion. When the staining results are evaluated, what will the digested section demonstrate?

A

Sites of glycogen removal.

86
Q

What is an example of a birefringent pigment?

A

A formalin pigment.

87
Q

Acid mucosubstances and neutral mucosubstances can be differentiated by staining with what dual stain?

A

Alcian blue and PAS.

88
Q

In the colliodal iron stain, the principal of the reaction is believed to be the formation of an ionic bond between the ferric ion with the carboxyl and sulfate groups of:

A

Acid mucosubstances.

89
Q

In order to suppress background and nonspecific staining, a Congo red solution frequently contains:

A

Sodium chloride.

90
Q

A lymph node is stained with a silver method for Retic and counterstained with nuclear fast red. After dehydration, there is a film over the entire slide that remains through clearing and coverslipping. What happened?

A

The slides were not adequately rinsed with water after counterstaining.

91
Q

A section of muscular artery had been stained with the Verhoeff-van gieson procedure. Microscopic evaluation shows blue-black nuclei, black elastic fibers with orange collagen and muscle. What happened?

A

The picric acid was too dilute so the muscle and collagen weren’t properly differentiated.

92
Q

Yellow-brown pigment, often found in cardiac muscle and liver cells in increasing amounts with age or debilitated states is known as:

A

Lipofucsin.

93
Q

Hemosiderin, hemoglobin and bile pigment are all classified as:

A

Endogenous pigments.

94
Q

The most appropriate hematoxylin solution for nuclear staining in a lengthy procedure that uses very acidic solutions is one that is mordanted with:

A

Iron, because it will not be removed by acids.

95
Q

A modified phosphotungstic acid-hematoxylin procedure can be used to demonstrate:

A

Glial fibers.

96
Q

What is an example of an argyrophil method?

A

The Grimelius stain.

97
Q

In the Bodian technique, interference with primary staining may occur with prolonged treatment in:

A

Oxalic acid will ruin the silver proteinate reaction in the Bodian technique.

98
Q

The staining intensity of eosin is increased in muscle and RBCs when tissues are fixed in:

A

Zenker because it does not bind to cationic groups.

99
Q

The lab has used all of the supply of aluminum hemaxotylin solution. In order to prepare a new solution for immediate use, the solution must be:

A

Chemically ripened (oxidation).

100
Q

When using the cresyl echt violet method, Nissl substance and nuclei can be preferentially stained by varying the degree of differentiation and the:

A

Solution pH.

101
Q

Delafield hematoxylin is most commonly aged with:

A

Light.

102
Q

What is an amphoteric substance?

A

An amphoteric substance can act either as an acid or as a base depending on the pH of the solution.

103
Q

Microscopic sections stained with H&E show a lack of nuclear staining even though the hematoxylin is well ripened and all of the other solutions are fresh. What happened?

A

The prolonged storage of the tissue in NBF.

104
Q

Microscopic review of an H&E stain shows that the eosin is very pale. What happened?

A

There was inadequate rinsing after neutralization.

105
Q

What is the staining mechanism in which dye adheres to the surface of structures?

A

ADsorption.

106
Q

When is Weigert hematoxylin considered unsatisfactory?

A

After three to four days.

107
Q

In a routine staining series, slides placed in xylene for clearing prior to coveslipping have an opaque appearance. What happened?

A

There was incomplete dehydration.

108
Q

Tissue sections were stained for the recommended time with H&E using Harris hematoxylin. A QC check shows pale nuclear staining. What happened?

A

The slides spent too long in the differentiating solution.

109
Q

T/F: Dyes used for nuclear staining are basic.

A

True.

110
Q

A stock solution of Harris hematoxylin is being prepared for use in the routine H&E stain. To make it an effective stain, what must first happen to the heamtoxylin?

A

It must be oxidized (primed).

111
Q

What is an antigenic determinant present on a complete antigenic molecule?

A

Epitope.

112
Q

What is a dye that absorbs light and then emits its own light at a longer wavelength?

A

Flurochromes.

113
Q

What is a fungus that can be demonstrated with various stains for acid mucins?

A

Cryptococcus neoformans.

114
Q

ATPase stains are done on a muscle biopsy at pH 9.4, 4.6 and 4.3. The slides for pH 4.6 shows differentiation of type I and type II fibers but NOT type IIA and IIB fibers. What happened?

A

The pH should have been modified on the 4.6 stain.

115
Q

What is the pathologic condition characterized by abnormal deposits of iron in the liver?

A

Hemochromatosis.

116
Q

What is an enzyme technique that will demonstrate denervated muscle fibers and motor end plates?

A

alpha-naphthyl acetate esterase.

117
Q

What procedure will demonstrate H. Pylori without the use of a microwave oven?

A

The Diff-Quik.

118
Q

A research project requires the demonstration of lysosomes on frozen sections of muscle tissue. What stain should be used?

A

Acid phosphatase.

119
Q

In the PAS reaction, how is the development of the final product achieved?

A

Restoration of the quinoid structure.

120
Q

Mucicarmine positive material is noted in the cytoplasm of poorly differentiated malignant cells. What kind of cancer would this indicate?

A

Adenocarcinoma because mucicarmine is specific for epithelium.

121
Q

What is the rare enzyme technique that can be performed on paraffin sections?

A

The naphthol AS-D chloroacetate esterase.

122
Q

How will the activity of succinic dehydrogenase be destroyed?

A

Fixation.

123
Q

What are structures that speed up the rate of enzymatic reactions?

A

Cofactors.

124
Q

What are enzymes that catalyze chemical reactions in biological systems?

A

Proteins.

125
Q

What is a procedure that demonstrates some carcinoid tumors as well as the alpha pancreatic cells?

A

The Grimelius stain.

126
Q

A patient is suspected of having McArdle disease which is a glycogen storage disease. The disease is BEST demonstrated with which stain?

A

The phosphorylase technique.

127
Q

Experiments have shown that a particular lipid fraction known as the mycolic acid exists within the cell wall of what bacteria?

A

Mycobacterium tuberculosis.

128
Q

Ab molecules can belong to one of 5 immunoglobulin classes. What is the Ab class most commonly used in IHC?

A

IgG.

129
Q

What causes the red-brown nuclear staining sometimes seen with hematoxylin?

A

The solution has begun to breakdown.

130
Q

How can the excess background staining be broken down with IHC stains?

A

Pretreating with nonimmune serum from the same animal species as the secondary Ab.

131
Q

What fungi may appear as a mixture of budding yeast cells and pseudohypha elements in infected tissue?

A

Candida albicans.

132
Q

What are mycotic diseases that go beyond superficial or cutaneous involvement to affect vital organs called?

A

Systemic.

133
Q

In a suspected case of alzheimers, what stain would be ordered?

A

A Bielschowsky stain.

134
Q

What method would be used for the demonstration of rickettsias?

A

Giemsa.

135
Q

What does the succinic dehydrogenase technique stain?

A

Mitochrondria.

136
Q

What would make a good control for Ab Cam 5.2?

A

A section from a carcinoma.

137
Q

A rhabdomyosarcoma is suspected in a biopsy submitted to the lab. To aid in making a definitive diagnois, what stain would be ordered?

A

A PTAH stain would be ordered.

138
Q

In lung tissue, coated asbestos fibers can be made more visible by staining with which stain?

A

The Prussian blue reaction.

139
Q

What kind of cell is responsible for immunoglobulin production?

A

Plasma cells.

140
Q

Melanosis coli is a condition characterized by abnormal deposits of pigment that will give a positive reaction to which stain?

A

Schmorl.

141
Q

Astrocytes can be demonstrated by using the Ab that is specific for what IHC test?

A

GFAP.

142
Q

`What would be the best stain to demonstrate loss of muscle striations caused by dystrophic change?

A

The Mallory PTAH stain.

143
Q

What is the amorphous transparent gel- like material that forms the bulk of extracellular content in connective tissue?

A

The ground substance.

144
Q

What are the small eosinophilic inclusions found in the cytoplasm of neurons in patients infected with the rabies virus?

A

Negri bodies.

145
Q

What is a fungal disease characterized by narrow-based budding and carminophilia of yeast organisms?

A

Cryotpcoccus.

146
Q

What is a maltese cross?

A

It is a configuration that is produced in tissue sections by the polarization of talcum powder, or starch in general.

147
Q

What is the pigment occurring in Plasmodium parasites that is closely related to formalin?

A

Malarial.

148
Q

In IHC reactions using horseradish peroxidase, how is the solution of hydrogen peroxide in methanol used?

A

To block endogenous peroxidase.

149
Q

In ICH staining of NBF fixed tissue, does the heat-induced epitope retrieval enhance primary staining?

A

Yiss.

150
Q

Following IHC staining, both the positive control and the specimen show weak staining. What happened?

A

The epitope retrieval was incorrectly done.

Histology HTL Exam- BOC Study Guide (15 decks)

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