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Meducina > Module 1: Methods of Cytology and Histology > Flashcards

Module 1: Methods of Cytology and Histology Flashcards

1
Q

Collects and focuses a cone of light that illuminates the tissue slide on the stage

A

Condenser

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2
Q

Most commonly used staining dyes?

A

Combi of Eosin & Hematoxylin

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3
Q

Fixative of choice for Light Microscopy?

A

Buffered solution of isotonic formaldehyde (formalin)

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4
Q

Cellular parts that Eosin stains? What color is produced?

A

a. Stains other cytoplasmic stx b. Collagen Pink

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5
Q

What cellular antigens can IHC target?

A

a. Cytoplasmic b. Nuclear d. Cell membrane e. Lipid F. Proteins

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6
Q

What is the maximum resolving power in light microscopy?

A

0.2 um (1000-1500 times)

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7
Q

Cellular parts that Hematoxylin stains? What color is produced?

A

a. DNA in cell nucleus b. RNA-rich portion of the cytoplasm c. Matrix of cartilage Dark blue/purple

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8
Q

What are the complex solution for cell& tissue culture

A

a. Salts, amino acid, vitamins b. Serum & specific growth factor

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9
Q

Why the need section/ slice tissue samples?

A

Inorder to obtain thin translucent sections for the light to pass through and be visualized

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10
Q

What is currently used in tissue processing?

A

Automated tissue processor

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11
Q

What is Histology?

A

Study of TISSUE arrangements of the body to constitute organs

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12
Q

Most common procedure done in histologic research

A

Preparation of tissue slices. (Can be examined visually with transmitted light)

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13
Q

What does antigens produced by viruses diagnose?

A

Specific virus infections

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14
Q

This process uses microtome to slice & obtain tissue sample based on size for a particular visualization method

A

Sectioning

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15
Q

General Methods used in studying histology?

A

a. Microscope b. Molecular method

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16
Q

A phenomenon where substances irradiated by light of a proper wavelength emit light with a longer wavelength

A

Fluorescence

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17
Q

True or false, phase contrast microscopy can be used with living cultured cells

A

True

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18
Q

This pattern makes use of basic dyes and stain structure BLUE?

A

Basophilic

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19
Q

They are initiated directly from the cells, tissues and organs of animals/ human typically used for experiment for a few days.

A

Primary cell culture

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20
Q

Measurement of Tissue section for Electron microscopy

A

< 1 microns

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21
Q

Infiltration of tissue sample by placing it into a melted parrafin

A

Infiltration

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22
Q

Antigen found in nucleus

A

Estrogen receptors Progesterone receptors

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23
Q

T/F, cytology general involves looking at a single cell type

A

True

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24
Q

T/F, primary cell culture propagated repeatedly gives rise to cell lines

A

True

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25
Q

Steps in Tissue preparation?

A

a. Fixation b. Dehydration c. Clearing d. Infiltration E. Embedding

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26
Q

What structure are acidophilic?

A

Mitochondria Collagen Secretory granules

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27
Q

How does cell create an organism?

A

Cells + assoc. matrix (highly specialized & functionalized) —> Tissue (Fundamental)–> Organs –> System –> organism

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28
Q

T/F, in situ hybridization can be used to identify the cells containing specific messenger RNAS (MRNA) in which the corresponding gene is being transcribed

A

True

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29
Q

Collects & focuses a cone of light that illuminates the tissue slide on the stage

A

Condenser

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30
Q

Clinical applications cell & tissue culture

A

a. Used to study molecular changes that occur in cancer. b. Could analyze viruses, mycoplasma, & some protozoans c. Used to prepare tissues for routine genetic chromosomal analysis

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31
Q

This process infuses dyes to different cell type structures that has different staining pattern

A

Staining

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32
Q

Step of embedding where alcohol is removed thru miscible agents (Toluene & Xylene).

A

Clearing

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33
Q

What is the most common PROCEDURE used in histological research?

A

Preparation of tissue section to be studies in LIGHT MICROSCOPE

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34
Q

Placing a small piece of tissue in a solution of chemical fixatives that preserves by cross-lining & inactivating degradative enzyme

A

Fixation

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35
Q

T/F, TEM can magnify up to 400,000 times

A

True

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36
Q

How long does overall sample prep be ready?

A

12-2.5 days

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37
Q

What are fixatives?

A

Solution of stabilizing/ cross-linking compounds

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38
Q

How does tissue processing occur?

A

Tissue sections are processed into thin sections which is best for microscopy

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39
Q

What does specific cytokeratin diagnose

A

Tumors of epithelial origin

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40
Q

What is used to embed tissue?

A

Melted paraffin

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41
Q

What are the 2 basic staining pattern?

A

a. Basophilic SP b. Acidophilic SP

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42
Q

A single dye applied separately to distinguish additional features of a tissue

A

Counter-stain (i.e Eosin)

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43
Q

Appearance of highly oriented molecules (birefringence) under polarizing light microscopy

A

Bright red or Yellow Apple green- amyloid

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44
Q

A type of Electron microscopy that provides high resolution view of surfaces of cells, tissues, and organs

A

Scanning electron microscopy (SEM)

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45
Q

This exposes tissue for sectioning or slicing on a microtome

A

Trimming

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46
Q

Substances that exhibits Birefringence

A

Cellulose Collagen Microtubules Actin filament

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47
Q

Study of the tissues of the body and how they are arranged to constitute organs

A

Histology

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48
Q

A light wave vibrating in more than one plane

A

Unpolarized light

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49
Q

A method of localizing newly synthesized macromolecules in cells or tissue sections

A

Autoradiography

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50
Q

Utilizes the hexose rings of polysaccharides and other carb-rich tissues and stains each macromolecules distinctly

A

Periodic acid-Schiff (PAS) reaction

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51
Q

A method of using cover slip to completely cover the tissue section and subject it to mounting medium

A

Mounting

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52
Q

This pattern makes use of acidic dyes and stains structure PINK

A

Acidophilic sp

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53
Q

Why use paraffin in tissue infiltration?

A

It makes tissue sample more rigid and easier to cut

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54
Q

What is an Ideal microscopic preparation?

A

a. Should be well-preserved tissue b. Has the same structure as the molecular composition in the body

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55
Q

Processes of embedding

A

a. Dehydration b. Clearing c. infltration

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56
Q

Resolution of TEM microscope

A

3 nm

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57
Q

True or False, In fluorescence microscopy, the emission of light upon tissue irradiation is in the visible portion of the spectrum.

A

True

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58
Q

Acidophilic stains?

A

a. Eosin b. Orange G c. Acid fuchsin

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59
Q

How to get total magnification?

A

Magnification power of objective lense x Magnification power of ocular lense

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60
Q

This uses a lens system that produces visible images from transparent objects

A

Phase contrast microscopy

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61
Q

The process of transforming unpolarized lt. to polarized lt.

A

Polarization

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62
Q

Explain direct method of IHC

A

a. Makes use of a single Ab b. Application of reagent containing specific Ab for binding to target antigen (Anti-A) c. Development of Antigen-Ab reaction indicates presence of target antibody d. Color change upon reaction indicates the presence of target antigenp

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63
Q

Enzymes that can be detected using histochemistry:

A

a. Phosphatase b. Dehydrogenase c. Peroxidase

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64
Q

How tissues are collected for histological study

A

a. Scraping or brushing the tissues (papsmear) b. Collecting body fluids (urine / resp secretion) c. Fine needle aspirations (abdominal fluid: ascites, pleural fluid: lungs, CSF: spinal canal)

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65
Q

Examination of a single cell type as often found in fluid specimen.

A

Cytology

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66
Q

Fixative of choice for Electron microscopy?

A

Glutaraldehyde solution followed by buffered Osmium tetroxide

67
Q

A process of fixing tissue-embedded wax piece into a supporting wooden block (plastic embedding cassette) to be fixed for sectioning to microtome

A

Blocking of sections

68
Q

Importance of Fixation

A

a. Prevents Auto-lysis b. Prevents bacterial degradation of Tissue c. Preserve cell & tissue structure

69
Q

Difference of typical staining procedure vs IHC

A

In IHC, no staining medium is incorporated but the Ab-antigen complex will stain the slide

70
Q

Temperature needed to maintain melted paraffin

A

52~60 C

71
Q

What is Tissue?

A

Cellular organizational level in-between a cell & complete organs. Group of similar cells with their EC matrix from the same origin & carry out specific fxn.

72
Q

Optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out of focus light (scattered light) in image formation

A

Confocal microscopy

73
Q

Visual markers used in IHC

A

a. Fluorescent dye b. Colloidal metal c. Hapten d. Radioactive markerw e. Enzyme (peroxidase)

74
Q

They act as catalyst to bring about a specific type of biochemical reaction

A

Enzyme

75
Q

A microscopy method based on the interaction of tissue components with beams of electrons.

A

Electron microscopy

76
Q

A technique in which a single stranded DNA or RNA sequence called probe is allowed to form complementary base pairs with DNA or RNA present in a tissue or chromosome sample

A

In situ hybridization

77
Q

T/F, usual cut used in TEM is 40-90nm

A

True

78
Q

Fixative for electron microscopy

A

Glutaraldehyde / formaldehyde

79
Q

A microscopy method which uses polarized light to visualize highly organized subunit

A

Polarizing microscopy

80
Q

Two main types of electron microscope

A

a. Transmission electron microscope (TEM) b. Scanning electron microscope (SEM)

81
Q
A
82
Q

A method for localizing cellular structures using a specific enzymatic activity present in the structures

A

Enzyme histochemistry

83
Q

Breast duct cell tumor can be diagnosed by what specific antigen?

A

Steroid hormone receptor

84
Q

Magnification used to observe high magnification of more detailed areas

A

X40

85
Q

Type of Ab-antigen binding

A

Direct

86
Q

Determining the localization of a gene in a specific chromosome cannot be applied in In situ hybridization

A

False In situ is ideally applied

87
Q

Describe extracellular matrix?

A

Contains many kinds of molecules (Collagen)

88
Q

Identify this staining in IHC [insert pic]

A

Cytoplasmic staining

89
Q

Antigen found in Cytoplasm

A

Cytokeratin (present in epith cells)

90
Q

In autoradiograph, they indicate region of cells in which specific macromolecules where synthesized prior to fixation

A

Silver grains

91
Q

A machine used for tissue embedding & sectioning

A

Tissue processor

92
Q

Measurement of Tissue section for Light microscopy

A

1-10 microns

93
Q

Commonly used visual markers for IHC light microscopy

A

Peroxidase (enzymes)

94
Q

A microscope that has a strong UV light source w/ special filters that select traces of different wavelengths emitted by a certain substance

A

Fluorescence Microscopy

95
Q

Methods for localizing cellular structure in tissues using enzyme activity present in these structure

A

Histochemistry & cytochemistry

96
Q

How does manual tissue processing works?

A

Tissues usually undergo chains of reaction

97
Q

Objective of trimming tissues?

A

To expose full face of tissue section to microtome knife

98
Q

What to with the paraffin infiltrated tissue?

A

Placed in a tissue block with melted paraffin then hardened.

99
Q

This magnification is used for observing a large area field at low magnification

A

X4

100
Q

What does protein polypeptide hormone diagnose?

A

Certain endocrine tumors

101
Q

How does Glutaraldehyde & Formaldehyde prevent degradation?

A

Interaction with Amine groups of Proteins

102
Q

This method reveal locations of newly synthesized glycoproteins containing a specific sugar

A

Autoradiography

103
Q

What is cytology?

A

Study of STRUCTURE & FUNCTION of cell

104
Q

What color does PAS reaction produce?

A

Purple or magenta

105
Q

Clinical application of cytology

A

a. Used to dx or screen for CA b. Screen for fetal abnormalities c. Papsmears to dx infectious organisms

106
Q

Who selects tissue sections?

A

Pathologist or medical officer

107
Q

Explain the indirect method of IHC

A

a. Makes use of two Ab b. Ab1 binds to target Antigen c. Ab2 binds to Ab1 d. Color change upon reaction indicates the presence of target antigenp

108
Q

Identify this staining in IHC [insert pic]

A

Membranous staining

109
Q

Combines histologicals, immunologicals, & biochemical techniques for identification of specific tissue component by means of specific antigen/ antibody tagged with a visible label

A

IHC

110
Q

T/F, in SEM, surface is coated with a very thin layer of heavy metal

A

True

111
Q

What are the basic steps of tissue processing?

A

A. Fixation of specimen B. Selection of tissue processing C. Processing of tissue D. Tissue embedding E. Blocking of tissue section F. Trimming G. Section cutting H. Routing & staining w/ hemotoxylin & eosin I. Mounting

112
Q

What embedding materials used? & where are they used?

A

a. Paraffin: light microscopy b. Plastic resins: light & electron microscopy

113
Q

How does electron microscopy allows for 1000 fold increase in resolution?

A

The wavelength in an electron beam is much shorter than that of light

114
Q

Refers to structures being observed under electron microscopy

A

Ultra structures

115
Q

Colors produced in electron microscopy

A

Black, white, shades of gray *lighter region- area less dense (easy electron passage) *darker region- area highly dense

116
Q

Magnification used to observe medium magnification & smaller field

A

X10

117
Q

Identify this staining in IHC [insert pic]

A

Nuclear staining

118
Q

What is the specimen in phase contrast microscopy?

A

Unstained cells and tissue section

119
Q

What should be obtained in section cutting?

A

A thin slice of tissue less than one cell layer thick w/o changing its architecture of tissue section.

120
Q

What protein is readily stained by Basic dye?

A

Nucleic acid

121
Q

What is tissue embedding?

A

Tissue infiltration with embedding materials to impart rigid consistency to the tissue

122
Q

Cultures of animal cells that can be propagated repeatedly & sometime indefinitely, also arises from primary cell culture

A

Cell line

123
Q

Type of Ab-Antigen binding

A

Indirect

124
Q

A special cutting device for tissue preparation

A

Microtome

125
Q

Resolution of TEM

A

3 nm resolution

126
Q

Implies the specific binding between two single strands of nucleic acid which occurs under appropriate condition if the strands are complimentary

A

Hybridization techniques

127
Q

Commonly used mounting medium?

A

D.P.X (distrene, plasticizer, xylene)

128
Q

What are the two interacting component of tissues?

A

Extracellular matrix & cell

129
Q

Functions of EC matrix

A

a. Supports the cell b. Contains fluid that transport nutrients to the cell and carry away waste & secretory products

130
Q

Why record tissue specimen?

A

Prevent specimen mixed up

131
Q

Objective of Tissue embedding

A

Facilitate sectioning by embedding tissue in a solid medium.

132
Q

A method for localizing specific antigen in tissues or cells based on antigen-antibody reaction

A

Immunohistochemistry (IHC)/ Immunocytochemistry

133
Q

Light waves vibrate in a single plane

A

Polarized light

134
Q

What does carcinoembryonic antigen diagnose?

A

Glandular tumors

135
Q

Application of polarizing microscopy

A

Myelin Osteoid Crystals

136
Q

A critical factor in obtaining a crisp detailed image w/ light microscope

A

Resolving power

137
Q

How does Tissue specimen recorded?

A

a. By numbering them like DBH/1256 b. BY RECORDING them in register

138
Q

Method of removing cells by drawing them thru a fine needle

A

Method of removing cells by drawing them thru a fine needle

139
Q

Live cells and tissue are maintained and studied outside the body

A

Cell and tissue culture

140
Q

T/F, exam of the entire tissue block

A

True

141
Q

Fixative for light microscopy

A

Buffered isotonic solution of 37% formaldehyde

142
Q

Enlarge & project the illuminated image of the object toward the eyepiece

A

Objective lense

143
Q

Smallest distance b/n two particles in w/c they can be seen as separate objects

A

Resolving power

144
Q

Color produced in peroxidase reactions

A

Brown

145
Q

T/F, in SEM, you can only visualize surfaces as beam does not pass thru specimens

A

True

146
Q

True or false, phase contrast microscopy follows the principle that light changes its speed when passing through the cellular and extracellular structures with different refractive indices

A

True

147
Q

A microscopy technique that Uses high intensity light often from a laser

A

Confocal microscopy

148
Q

What is fixation?

A

Placing a small piece of tissue in a solution of chemical fixatives that preserves by cross-lining & inactivating degradative enzyme

149
Q

Most commonly used microscopic method?

A

Brightfield microscopy

150
Q

Microscopy method that allows examination of cells without fixation or staining hence prominently used in all cell culture laboratories. It can also be used for living cells

A

Phase-contrast microscopy

151
Q

How does labeled antibody in IHC visualized?

A

Using light microscopy in the presence of a suitable colorogenic substrate system

152
Q

It will magnify the image for another x10 & project it to the viewer

A

Eyepieces or ocular

153
Q

Procedures done in IHC staining

A

[insert pic taken from module 1 part 3, 10:09]

154
Q

Final step before microscopic observation of tissue

A

Mounting a protective glass cover on the slide with clear adhesive

155
Q

More commonly used Immunohistochemistry method? Why

A

Indirect Labeling the protein of interest is amplified by indirect method

156
Q

Why does fixation carried out ASAP?

A

To prevent putrefaction & autolysis

157
Q

Step of embedding w/c transfers tissue to a series of increasing alcohol concentration until 100% to fully remove all water

A

Dehydration

158
Q

Example of Basophilic dye?

A

a. Toluidine Blue b. Alcian blue c. Methylene blue d. Hematoxylin

159
Q

Heavy metal used for surface coating in SEM (and how?)

A

Osmium tetroxide Lead citrate Bind to macromolecules increasing electron density & visibility

160
Q

Ability to rotate the direction of polarized light

A

Birefringence

161
Q

Produces an image of living cells with a more apparent 3D aspect.

A

Differential interference contrast microscopy (DICM)

162
Q

T/F, in situ hybridization is ideal for determining if a cell has a specific sequence of DNA such as a gene or part of a gene

A

True

163
Q

Antigen found in membrane staining in IHC

A

HER2/ neu

Meducina (6 decks)

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