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Lab Test > Direct Microscopic Somatic Cell Count > Flashcards

Direct Microscopic Somatic Cell Count Flashcards

1
Q

What is the maximum time unpreserved samples can be tested after initial collection?

A

72 hours

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2
Q

What are the preservatives that allow samples to be tested up to 7 days after collection?

A

0.02% 2-bromo-2-nitropropane-1,3-dio (Bronopol™) or 0.05% potassium dichromate (K2Cr2O7)

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3
Q

What is the required face velocity for a functional fume hood?

A

100 ft/min

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4
Q

What are the dimensions of the clean microscope slides used in the procedure?

A

2.54 x 7.62 cm

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5
Q

What is the diameter of the areas delineated on the microscope slides?

A

11.28 mm

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6
Q

What is the purpose of the metal syringe in the process?

A

For rapid and convenient transfer of 0.01 mL of milk

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7
Q

What is the acceptable weight delivery range for the metal syringe?

A

0.0103 ± 0.0005 g

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8
Q

What must be done annually for the fume hood?

A

Check, maintain records, and tag unit

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9
Q

What identification is required for the metal syringe?

A

Etched identification or imprinted serial number

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10
Q

Fill in the blank: Samples may be run up to _____ after initial collection if preserved.

A

7 days

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11
Q

True or False: Micropipettors are used for transferring more than 0.01 mL of milk.

A

False

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12
Q

What is the average weight to be checked for accuracy with the metal syringe?

A

0.0103 g

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13
Q

What is the significance of the accuracy check date for the metal syringe?

A

To ensure proper functioning and reliability

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14
Q

What is the average weight specified for accuracy check in item 6.e.4?

A

0.0103 +0.0005g

This is based on the average of 10 consecutive weighings with milk.

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15
Q

What should be done if using Artel PCS according to CP item 6.e.5?

A

See CP item 6.e.5

Specific instructions for using Artel PCS are outlined in that section.

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16
Q

What is required for the micropipettor used in the procedure?

A

Etched with identification (imprinted serial number acceptable); tag with accuracy check date

This ensures traceability and compliance with accuracy checks.

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17
Q

What type of needle is suitable for spreading milk film?

A

Dissecting Needle, Bent Point

This needle design facilitates even spreading of the film.

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18
Q

What temperature range should the drying device maintain?

A

40-45°C

This temperature range is crucial for effective drying.

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19
Q

What is required to monitor the temperature of the drying device?

A

Temperature measuring device

Accurate temperature monitoring ensures proper drying conditions.

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20
Q

What tools are required for dipping and holding slides?

A

Forceps or Slide Holder

These tools help in handling slides without contamination.

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21
Q

What should staining jars or trays have?

A

Tight fitting covers

This helps to prevent evaporation and contamination of solvents and stains.

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22
Q

What is the recommended storage condition for slides?

A

Clean, dust-free insect-proof boxes, cases or files

Proper storage conditions are necessary to maintain slide integrity.

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23
Q

What type of microscope is specified for use?

A

Binocular with 1.8 mm oil immersion objective, rack and pinion sub-stage, condenser with iris diaphragm

This microscope configuration is essential for detailed observation.

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24
Q

What ocular magnification is recommended for the microscope?

A

10X (12X or 12.5X), Huygenian or wide-field

These magnifications provide a suitable field of view for analysis.

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25
Q

What is the maximum Single Strip Factor (SSF) specified for the optics?

A

6070 or smaller

This ensures sufficient resolution for the analysis performed.

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26
Q

How often should each analyst measure field diameter and calculate SSF?

A

Annually

This regular check helps maintain accuracy in measurements.

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27
Q

To how many significant figures should the SSF be rounded?

A

Three significant figures

This precision is important for consistency in reporting.

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28
Q

What is the formula to compute the Single Strip Factor (SSF)?

A

SSF = 10,000/(11.28 x D)

D is the field diameter in mm measured using a stage micrometer.

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29
Q

What is the field diameter (D) used for in the calculation of SSF?

A

D is measured in mm using a stage micrometer.

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30
Q

What are the characteristics of a suitable mechanical stage for microscope examination?

A

Smooth action, does not drift, allows proper tracking of smears.

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31
Q

What is the purpose of the microscope lamp?

A

Provides adequate illumination.

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32
Q

What divisions are ruled on a stage micrometer?

A

0.1 mm and 0.01 mm divisions.

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33
Q

What is the refractive index of immersion oil?

A

1.51-1.52.

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34
Q

What is the first step in preparing the Levowitz-Weber Modification of the Newman-Lampert Stain?

A

Slowly add 0.6 g certified methylene blue chloride to 52 mL of 95% ethyl alcohol and 44 mL of tetrachloroethane in a 200 mL flask.

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35
Q

What safety precautions should be taken when making the stain?

A

Use gloves and prepare in a fume hood.

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36
Q

How long should the stain stand at the specified temperature?

A

12-24 hours at 4.5-7.5°C.

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37
Q

What type of filter paper should be used to filter the stain?

A

Whatman No. 42 filter paper or equivalent.

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38
Q

What should be added to the stain after filtering?

A

4 mL of glacial acetic acid.

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39
Q

How should the prepared stain be stored?

A

In a clean, tightly closed container.

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40
Q

What may cause problems with the stain when stored?

A

Traces of water or solvent.

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41
Q

What are alternative forms of the stain that can be used?

A

Commercially prepared (xylene or tetrachloroethane).

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42
Q

What information is required when using commercially prepared stains?

A

Brand, Lot #, Exp. Date.

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43
Q

What is Canadian Formula Stain?

A

Commercially prepared stain using xylene or tetrachloroethane

Includes lot number, expiration date, and brand information.

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44
Q

What are the components for preparing Alternate Methylene Blue Stain?

A

Combine:
* Cert. Methylene Blue Chloride (0.5 g)
* 95% Ethyl Alcohol (56 mL)
* Xylene (40 mL)
* Glacial Acetic Acid (4 mL)

Follow the preparation method outlined in item 14.

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45
Q

What is the fixative used for Pyronin Y-Methyl Green Stain for Goat, Sheep or Camel Milk?

A

Carnoy’s fixative

Components include chloroform, glacial acetic acid, and 100% ethyl alcohol.

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46
Q

What are the components for Carnoy’s fixative?

A

Combine:
* Chloroform (60 mL)
* Glacial Acetic Acid (20 mL)
* 100% Ethyl Alcohol (120 mL)

Can be prepared or commercially obtained.

47
Q

What is the preparation method for Pyronin Y-Methyl Green Stain?

A

Combine:
* Pyronin Y (1.0 g)
* Methyl Green (0.56 g)
* Water (196 mL)

Filter through Whatman No. 1 paper before use.

48
Q

How should Pyronin Y-Methyl Green Stain be stored?

A

Stain is light sensitive; store in brown bottle

Can also be commercially prepared.

49
Q

What are the steps for cleaning slides?

A
  1. Physically clean
  2. Soak new slides in strong cleaning solution
  3. Rinse thoroughly in flowing water (10-15 sec)
  4. Soak used slides in hot detergent or wetting agent until residues are removed; rinse as above

Ensures slides are free from contaminants.

50
Q

What is the recommended method for drying slides?

A

Air or heat dry with minimal exposure to dust, insects, etc. and store dry

51
Q

What should be done to slides before use if stored in alcohol?

A

Flame just before use

52
Q

How should each sample area on a slide be identified?

A

Legibly and indelibly identify each sample area on margin of slide

53
Q

What is the procedure for sample agitation?

A

Mix samples or subsamples by shaking 25 times in 7 sec with a 1 ft movement or vortex for 10 sec at maximum setting

54
Q

What is the time limit for using samples after agitation?

A

Use within 3 min

55
Q

What temperature can high fat samples be warmed to before testing?

A

40°C for no longer than 10 min

56
Q

What is the rinsing procedure for a metal syringe before use?

A

Rinse syringe 2-3 times in clean, 25-35°C tap water

57
Q

How deep should the syringe be inserted into the milk before transferring the test portion?

A

Not over 1 cm below surface of milk

58
Q

What should be done after rinsing the syringe with milk?

A

Fully depress and release plunger and withdraw test portion

59
Q

How should excess milk be removed from the syringe tip?

A

With clean paper tissue, wipe downward away from tip

60
Q

What is the correct technique for expelling the test portion onto the slide?

A

Touch the slide with the tip and expel the test portion

61
Q

When spreading the milk on the slide, what should be avoided?

A

Do not release plunger until after touching off and removing tip from slide

62
Q

What technique should be used to spread the milk on the slide?

A

Spread milk with point of bent needle point

63
Q

What should be done between preparing multiple smears?

A

Complete steps 21.a through 21.e.4 before starting the next smear

64
Q

At what temperature should smears be dried?

A

40-45°C within 5 min on level surface

65
Q

What should be avoided to prevent smears from cracking and peeling during staining?

A

Heating too rapidly

Rapid heating can cause damage to the smear, affecting the quality of the staining process.

66
Q

How should smears and slides be treated until read?

A

Protected from damage

Proper handling is essential to maintain the integrity of the samples.

67
Q

What should be done to prevent residues from drying on a metal syringe?

A

Clean immediately after use

Prompt cleaning helps maintain the functionality and hygiene of the instrument.

68
Q

What should not be removed from the syringe unless necessary?

A

Spring

The spring mechanism is critical for the operation of the syringe.

69
Q

What type of detergents should be used for cleaning syringes?

A

Soap-less detergents and/or fat solvents

These cleaning agents are effective without leaving harmful residues.

70
Q

How should residues be cleaned from the measuring tube of a syringe?

A

By circulating detergent with bulb on delivery end

This method ensures thorough cleaning of the measuring tube.

71
Q

What should be used to clean the piston of a syringe?

A

Dry paper tissue

Using dry tissue prevents moisture accumulation that could affect performance.

72
Q

What should be done with the micropipettor tip for each sample?

A

Use a new tip

This practice prevents cross-contamination between samples.

73
Q

What is the correct method for withdrawing a sample with a micropipettor?

A

Depress plunger, insert tip below surface, release plunger slowly

This technique minimizes the introduction of foam and bubbles.

74
Q

How should excess liquid be removed from the exterior of a micropipettor tip?

A

Wipe away from the tip with clean paper tissue

This prevents contamination of the sample being processed.

75
Q

Where should the micropipettor tip be placed when preparing a smear?

A

Near the center of the area for smear

Proper placement ensures an even distribution of the sample.

76
Q

What should be done if the pipettor has two stops when expelling sample?

A

Depress plunger to second stop, touch off

This ensures complete expulsion of the sample.

77
Q

What technique should be used to spread milk on the slide?

A

With the point of a bent needle, not hockey stick style

This technique allows for a more controlled and even smear.

78
Q

What should be done between samples when using a needle for spreading?

A

Wipe needle dry on tissue

This prevents contamination of subsequent samples.

79
Q

What is the recommended drying temperature for smears?

A

40-45°C

This temperature range ensures proper drying without damaging the smear.

80
Q

Within what time frame should smears be dried?

A

Within 5 minutes

Timely drying helps preserve the quality of the smear for analysis.

81
Q

What should be avoided to prevent smears from cracking and peeling during staining?

A

Do not heat too rapidly

82
Q

How should smears and slides be protected until they are read?

A

Protect from damage

83
Q

What is the first step in the staining process using Levowitz-Weber and Methylene Blue Stains?

A

Use ventilated hood for steps 24.a.2-4

84
Q

For how long should slides be submerged or flooded in stain during Levowitz-Weber and Methylene Blue Stains?

A

2 min

85
Q

What should be done to drain excess stain from the slide?

A

Rest edge of slide on absorbent paper

86
Q

What temperature should the tap water be for rinsing stained slides?

A

35-45°C

87
Q

What is the key characteristic of the Pyronin Y-Methyl Green Stain?

A

Stain is light sensitive

88
Q

List the steps in the staining scheme for Pyronin Y-Methyl Green Stain.

A
  • Carnoy’s Fixative
  • 50% Ethanol
  • 30% Ethanol
  • DI or MS Water
  • Stain
  • N-Butyl Alcohol
  • Xylene
89
Q

What is the duration for the Carnoy’s Fixative step in the Pyronin Y-Methyl Green Stain?

A

5 min

90
Q

What color do cells stain in the Pyronin Y-Methyl Green Stain?

A

Blue or blue-green

91
Q

What color do RNA and the background stain in the Pyronin Y-Methyl Green Stain?

A

Pink

92
Q

What should be done if smears will not adhere to slides after Carnoy’s fixative?

A

Allow slide to dry for approx. 10 min, protected from light

93
Q

What adjustment should be made to the microscope lamp during examination?

A

Adjust to provide maximal optical resolution

94
Q

What is the first step to locate the edge of the smear to be read?

A

Use low power

95
Q

What should be placed on the smear for examination?

A

1 drop immersion oil

96
Q

What is the first step when using an oil immersion lens?

A

Carefully lower oil immersion lens

97
Q

What should be focused on and located at the beginning of cell counting?

A

Center of edge of area

98
Q

How should cells be counted in a smear?

A

Count all cells in field wide strip across diameter of a single smear

99
Q

What type of cells possess a nucleus that stains dark blue in bovine secretory systems?

A

Somatic cells

100
Q

In which secretory systems do nuclei stain blue or blue-green?

A

Apocrine (caprine) secretory systems

101
Q

When counting cells, which cells should be included?

A

Cells touching one edge of the strip but not the other edge

102
Q

How should fragments be counted?

A

Only if more than 50% of the nuclear material is visible

103
Q

How should clusters of cells be counted?

A

Count as one unless nuclear units are clearly separated

104
Q

What should be done if there is doubt about counting a cell?

A

Do not count

105
Q

What should be recorded after examining each smear?

A

Record strip count

106
Q

How often should comparative counting between analysts be conducted?

A

Monthly

107
Q

What is the first step in reporting somatic cell counts?

A

Record of strip count for each smear examined

108
Q

How is DMSCC/mL computed?

A

Multiply number of cells counted (strip count) by the SSF

109
Q

What should somatic cell counts be reported as?

A

DMSCC/mL

110
Q

How should the first two left-hand digits of DMSCC/mL be recorded?

A

Round as necessary

111
Q

What rounding rule applies when the third digit is 5 and the second digit is odd?

A

Round up

112
Q

What rounding rule applies when the third digit is 5 and the second digit is even?

A

Round down

113
Q

What form is maintained for Direct Microscopic Somatic Cell Count?

A

FORM NCIMS 2400d

Lab Test (13 decks)

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