Module 1 [cont]: Methods of cytology and histology

Question 1

How many lenses does compound light microscope hae.

Answer 1

Two or more

Question 2

Light source for compound LM?

Answer 2

Transmitted light

Question 3

T/F, compound light mic. Uses live specimens such as examination of stool for parasites

Answer 3

True

Question 4

Total magnification of CLM?

Answer 4

Not more than 2000x

Question 5

Max magnification of CLM?

Answer 5

400x

Question 6

T/F: eye piece only enlarges the images obtain by the objective and doesnt improve the resolution of the image

Answer 6

True

Question 7

3 objectives of CLM & their magnifications

Answer 7

a. Scanning objective- 4x
b. Low power objective- 10x
c. High power objective -40x (blue line)

Question 8

Part of CLM where it serves as a platform upon which the slide rests

Answer 8

Stage

Question 9

A transparent lens through which light reveals. Size and brightness can be adjusted

Answer 9

Iris diaphragm

Question 10

Light source of the clm

Answer 10

Lamp

Question 11

Steps on using microscope (shortcut)

Answer 11

a. Power on, make sure light is working; objective is set to LPO or scanning
b. Always start at scanning objective
c. Place glass slide on stage; fix using specimen holder
d. Move the slide to focus on image you want to see
e. Adjustment of course adjustment & fine adjustment knob
f. Adjust light intensity
g. Once image is clear, you can switch to a higher magnification to see individual details
h. If finished, switch back to LPO
i. Remove slide carefully and put it in proper box
J. Turn off microscope

Question 12

Knob that lowers the objective lenses

Answer 12

Course adjustment knob

Question 13

Knob that gives clearer image

Answer 13

Fine adjustment knob

Question 14

T/F, to preserve the morphologic and chemical integrity of the cell in as life-like manner as possible is the primary aim of fixatives

Answer 14

True

Question 15

Size of a tissue block used in light microscopy

Answer 15

2 x 0.4 sq. cm

Question 16

Size of a tissue block used in electron microscopy

Answer 16

1-2 sq. mm

Question 17

Main factors involved in fixation

Answer 17

a. Hydrogen ion concentrion (6-8)
b. Temperature
c. thickness of sxn
d. Osmolality
e. Concn. of Fixative
f. Duration of fixative (6-24hrs)

Question 18

Process of removing intracellular and extracellular water

Answer 18

Dehydration

Question 19

What percent of alcohol is used to dehydrate delicate tissues (i.e embryonic tissues)

Answer 19

30%

Question 20

Most recommended alcohol for dehydration and best dehydrating agent

Answer 20

Ethanol

Question 21

Why do we need to dehydrate tissue samples?

Answer 21

To remove water from previous fixation where in melted paraffin that will be used to infiltrate the tissue subsequently is Hydrophobic

Question 22

Most used clearing agent

Answer 22

Xylene

Question 23

Most common embedding medium being used

Answer 23

Paraffin

Question 24

It is used to adhere tissue sections in an untreated glass slide

Answer 24

Bonding Agent/ Adhesive

Question 25

Two example of bonding agents

Answer 25

a. albumin | b. poly-l-lysine

Question 26

Most common bonding agent

Answer 26

Albumin

Question 27

Two methods of staining

Answer 27

a. direct staining | b. indirect staining

Question 28

A process of giving color to the given sections by using aqueous or alcoholic dyes

Answer 28

Direct staining

Question 29

A staining method wherein the action of dye is intensified by using a mordant which serves as a link between tissue and dye

Answer 29

Indirect staining

Question 30

T/F, nucleus is basic in nature hence stains blue by Hematoxylin

Answer 30

False, acidic

Question 31

T/F, Cytoplasm is basic in nature hence in stained by Eosin pink

Answer 31

True

Question 32

A dye of combination of an acid and basic dye

Answer 32

Neutral dye

Question 33

Example where Neutral dye is used?

Answer 33

Leishman staining

Question 34

It gives a protective cover over the smear and make a permanent bond between the coverslip and the slides

Answer 34

Mounting media

Question 35

Requirement for a mounting media

Answer 35

a. same refractive index as the coverslip and glass slide (1.52 -1.54) b. should be colorless c. quick drying and stick to the slide d. resist contamination particularly the growth of microbes e. miscible with clearing agent

Question 36

What structures can PAS stains?

Answer 36

a. glycoprotein b. mucin c. mucoprotein d. glycoprotein e. basement membranes f. capsules g. blood vessels

Question 37

Color stain of PAS for positive

Answer 37

Purple/ Red/Magenta

Question 38

Color stain of PAS for nuclei

Answer 38

Blue

Question 39

Modified PAS which can stain DNA of cell nuclei

Answer 39

Feulgen reaction

Question 40

Lipid soluble dyes that lipid rich structure (phospholipids and neutral fats)

Answer 40

Sudan black

Question 41

Color stain of Sudan black if positive

Answer 41

blue black

Question 42

Color stain of Sudan black for nuclei

Answer 42

red

Question 43

Uses silver salts to stain certain ECM fibers and specific cellular elements in nervous tissues

Answer 43

Metal impregnation

Question 44

What is produced in Metal impregnation?

Answer 44

opaque black deposit on bacteria or cell surface

Question 45

T/F, Light is either passed through or Reflected in Brightfield microscopy

Answer 45

True

Question 46

Other term for eyepiece

Answer 46

Ocular lens

Question 47

T/F, eyepiece or ocular lenses improves the resolution of the object

Answer 47

False, it does not improve but only enlarges the image

Question 48

Objective used for locating objects and scanning slides quickly

Answer 48

Scanning objective

Question 49

Color and magnification of line in scanning obj

Answer 49

red line- 4x

Question 50

color and magnification of line in Low power obj

Answer 50

yellow line- 10x

Question 51

color and magnification of line in high power obj

Answer 51

blue line- 40x

Question 52

Lens that allows you to find and center object in a slide

Answer 52

Low power object

Question 53

Lens used to zoom in for closer viewing

Answer 53

High power objective

Question 54

It determines the quality of the image, clarity, and richness of detail

Answer 54

Resolving power

Question 55

What is the fluorescent stain used?

Answer 55

Acridine orange

Question 56

T/F, acridine orange binds only to the DNA

Answer 56

False, both DNA and RNA

Question 57

Light emitted by DNA and RNA under fluorescence microscopy

Answer 57

DNA- Yellow | RNA- orange

Question 58

Specimen for Phase contrast microscopy

Answer 58

unstained cells and tissue sections

Question 59

uses lens that produces visible images from transparent objects

Answer 59

PCM

Question 60

Used to study cell cycle in live cell

Answer 60

PCM

Question 61

Digitally reconstructs the specimen into a 3D image; records ion movements; determines enzymatic activities in tissues

Answer 61

Confocal microscopy

Question 62

Used in detecting cardiac amyloidosis (apple green birefringence)

Answer 62

cardiac amyloidosis

Question 63

It can visualize individual viral particles, diagnosis of glumerulopathies

Answer 63

Electron microscopy

Question 64

Fixative for electron microscopy

Answer 64

Glutaraldehyde

Question 65

Cut object in TEM

Answer 65

40-90 nm

Question 66

T/F, in TEM, electrons pass directly through the specimen

Answer 66

true

Question 67

T/F. in SEM, electrons bounce off the surface of the specimen at an angle and 3D image is produced

Answer 67

true

Question 68

Complex solutions that is used to sustain cells and tissue culture

Answer 68

Salts, Amino acid, vitamins, serum and specific growth factor

Question 69

Refers to as low passage or finite cell lines, aka diploid

Answer 69

Established cell line

Question 70

T/F, established cell lines can be maintained longer than primary cell line

Answer 70

True

Question 71

Altered cells that can be maintained indefinitely, the cells are heteroploid

Answer 71

Continuous cell lines

Question 72

W1-38, MRC-5, IMR-90 are examples of what cell line

Answer 72

Established cell line

Question 73

Hela, Hep-2, A549 and vero cells are example of what cell lines

Answer 73

Continuous cell line

Question 74

What is used in autoradiography as microdetector

Answer 74

Silver bromide crystals

Question 75

In autoradiography, they indicate the cells or regions of cells where macromolecules are synthesized

Answer 75

Silver grains

Question 76

Technique used to identify diagnostically useful enzyme or other tissues

Answer 76

Enzyme histo/Cytochemistry

Question 77

T/F, histochemistry and cytochemistry serves to detect early metabolic changes in biopsies and autopsy tissues

Answer 77

true

Question 78

T/F, immunohistochemistry, enables us to visualize the distribution and localization of specific cellular components within cell or tissue

Answer 78

true

Question 79

The most important requirement of Immunohistochemistry is the availability of an antibody against the protein to be detected

Answer 79

true

Question 80

A procedure in IHC wherein it uses heat , couple with specific buffered solutions to recover antigen reactivity

Answer 80

Antigen retrieval

Question 81

Procedure for IHC

Answer 81

(insert photo)

Question 82

Clinical application for IHC

Answer 82

useful in cancer diagnosis

Question 83

Allows a specific identification of sequences in genes or RNA

Answer 83

hybridization technique