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Clinical Embryologist's exam samples. > ART-Laboratory procedures > Flashcards

ART-Laboratory procedures Flashcards

1
Q

Regarding oocyte assessment two anomalies remain that should be handled with caution:

A

Regarding oocyte assessment two anomalies remain that should be handled with caution:

  • (i) ‘giant’ oocytes because of their likely abnormal` genetic constitution and
  • (ii) sER clusters because of their potentially lethal outcomes.
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2
Q

The time for syngamy check is different for ICSI And IVF Insemination

A

true

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3
Q

The nature of the zona pellucida is important in oocyte grading.

A

false

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4
Q

Fast Cleaving embryos are considered normal

A

False

Fast Cleaving embryos are considered at risk of being abnormal

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5
Q

A fragmentation of 25% in a 4cell embryo equates to one blastomere in volume

A

true

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6
Q

A poor cleaving embryo may have either:…… , …….., …….

A
  • severe fragmentation
  • cells out of phase
  • multinucleation
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7
Q

A 4AA Grade and a 411 Grade Blastocyst Are the Same?

A

true

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8
Q

Following aspiration, human oocytes are best held in

  1. Follicular fluid until fertilization at body temperature
  2. Bicarbonate buffered media at room temperature
  3. HEPES or MOPS buffered media at room temperature
  4. HEPES or MOPS buffered media at 37ºC
A

4.

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9
Q

If Day 0 is time of ovulation. human embryos would naturally enter the uterus on:

  1. Day 1
  2. Day 2
  3. Day 3
  4. Day 4
  5. Day 5
A

4.

Embryo enter the Uterus d4 of ovulation in natural

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10
Q

Which Statement is NOT true?

  1. pH strips are useful for checking
  2. Phenol red is a pH indicator and potentially oestrogenic
  3. Phenol red turns yellow in media with a pH of 6.5
  4. pH strips are usually sold non-sterile
  5. Phenol red is a clear solution that turns red in culture media
A

5.

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11
Q

The Optimum Embryo Development Stage for Transfer is:

  1. Always the blastocyst stage as these are the stronger embryos
  2. What works best in your own clinic
  3. Cleavage stage embryos since some grading is possible
  4. Whatever the patient requests
  5. When the embryonic genome is activated
A

2.

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12
Q

In ICSI, Immotile Sperm may be used Successfully after:

  1. positive response following exposure to hypo-osmotic medium
  2. electroporation
  3. treatment with calcium ionophore A23187
  4. showing motility in pentoxifylline containing medium
A

1, 4

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13
Q

To increase Humidity in Incubator……..

A

Placing a tray with 1L of water in the incubator will create humidity levels > 75%

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14
Q

advantages of oil overlay in culture media…….

A

reduce changes in Temp- and PH

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15
Q

Sequential Media was developed because there are changes in………………..

A

Sequential Media was developed because there are changes in embryo metabolism during cleavage

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16
Q

High O2 Coc- may increase the……….

A

free radical formation

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17
Q

Mineral Oil and Paraffin Oil

A

Paraffin oil and mineral oil are chemically slightly different

The polycarbon lipid tail of mineral oil contains more unsaturated bonds than paraffin oil

This makes mineral oil more unstable and prone to attack by free oxygen radicals and photo-oxidation

Paraffin oil is derived from fractionated distillation

Silicone Oil is a polymer

Silicone Oil may be more stable that paraffin oil.

Silicon oil is derived from gas chromatogrphy

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18
Q

High Temperature may be detrimental to the Oil

A

true

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19
Q

Washing Media Before use is a good idea because….

A
  • Batch To Batch Variability
  • It May Reduce Or Remove Water Soluble Toxins
  • washing Media Containing EDTA may Reduce or Remove Cations Such as Zinc
    *
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20
Q

Some Mineral Oil may Contain Preservatives Such as

A

Alpha-Tocopherol

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21
Q

Fertilisation Media Not Contain EDTA because:

A
  • EDTA may chelate Ca2++
  • Ca2++ is requried for the acrosome reaction
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22
Q

Biopsy Media free of Ca2++ but contain EDTA because….

A

to perform biopsy ⇒►and reduce blastomere compaction

Ca2++ is required for compaction

If Ca2++ is absent, the blastomeres cannot adhere

► then it can be biopsied

EDTA is a chelating agent so it will remove Ca2++ and facilitate biopsy

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23
Q

The Oocyte has …………..requirement for glucose

A

little or limited

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24
Q

Glucose is added to fertilization media for…….

A

sperm metabolism and motility

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25
Q

Penicillamine is sometimes used as a replacement for EDTA

A

true

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26
Q

Zinc and Cadmium are two potentially toxic cations removed by EDTA

A

true

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27
Q

EDTA is also used as a Free Radical Scavenger

A

true

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28
Q

EDTA can …………..Glycolysis in early cleavage stage Embryos

A

suppress

29
Q

protein sources supplementation of culture media used in early years….

A
  • Bovine serum albumin
  • Human fetal cord serum
  • Fertile Human donor serum
  • Maternal Human serum

The main problems in their use include:

Occult toxicity

Immense lack of consistency

Variation in the source and grading

Untraceable risk of disease transmission

30
Q

The development of stage specific media formulations allowed:

A

Replacement of serum with Human Serum Albumin (HSA)

Development of serum substitutes to augment HSA

The substitutes were mainly serum globulins

31
Q

Pregnancy has been achieved in protein free media that includes:

  • Polyvinvyl pyrrolidone (PVP)
  • Polyvinyl alcohol (PVA)
  • Hyaluronic acid
A

true

32
Q

The requirements for HSA And Supplements such as Globulins and Growth Factors Appear Stage Dependent.

A

true

33
Q

Phenol Red is

A

a guide to media pH

Not toxic to embryos

Not toxic to sperm

Not toxic to eggs

Straw colour at pH 7.2

Yellow at pH 7.0

Red at pH 7.4

Maroon at pH 7.8

34
Q

The Actual PH Of Culture Medium With 5%-6% CO2 will be influenced by:

  1. Protein concentration
  2. The type of protein Supplementation
  3. Altitude
  4. Age of the medium
  5. The number of times a bottle has been opened
  6. The latitude
  7. The longitude
A

1.

35
Q

A Change in the color of culture media is most likely due to

  1. The age of the medium
  2. The number of times the bottle has been opened
  3. The loss of CO2 from the medium
  4. Contamination
  5. Infection
A

2, 3

36
Q

The Shelf Life of CM (at 5˚C) is:

  1. 2 weeks
  2. 4 weeks
  3. 6 weeks
  4. 8 weeks
  5. 10 weeks
  6. 12 weeks
A

5.

The Shelf Life of culture media CM (at 5˚C) is 10 weeks

37
Q

The Main Source of a buildup of ammonia in culture media is due to

  1. Glucose
  2. Pyruvate
  3. Bicarbonate
  4. Glutamate
  5. Glutamate dipeptide
  6. Non-essential Acids
A

4.

38
Q

The ability of an embryo to regulate its metabolism is directly related to its ability to implant and form a viable pregnancy

A

true

39
Q

There are suggestions growth that rapid cleavage leads to loss of viability

A

true

40
Q

precocious exposure to glucose may cause the early cleavage embryo to divide too fast leading to loss of energy stores and viability

A

true

41
Q

The Main Metabolic Pathways Include

A

Pyruvate and lactate

Glucose

Amino Acids

Fatty acid and b-oxidation

42
Q

The Oviduct concentration of pyruvate is:

  1. 0.22 mM
  2. 0.32 mM
  3. 0.42 mM
  4. 0.52 mM
  5. 0.62 mM
A

2.

43
Q

The Uterine concentration of pyruvate is:

  1. 0.1 mM
  2. 0.2 mM
  3. 0.3 mM
  4. 0.4 mM
  5. 0.5 mM
A

1.

44
Q

The Oviduct Concentration of LACTATE is:

  1. 6.5 mM
  2. 7.5 mM
  3. 8.5 mM
  4. 9.5 mM
  5. 10.5 mM
A

5.

45
Q

Oviduct natural environment

Uterine

Pyruvate conc-

  1. 32 mM
  2. 1 mM

LACTATE conc-

  1. 5 mM
  2. 87 mM

Glucose

  1. 5 mM
  2. 15 mM

Amino Acids

non-essential amino acids

Mix of ess & non-ess

A
46
Q

The Metabolic Quotient of an early-stage cleavage embryo is similar to

  1. heart tissue
  2. brain tissue
  3. muscle tissue
  4. liver tissue
  5. tumor tissue
A

2.

47
Q

The Metabolic Quotient of a blastocyst is similar to

  1. heart tissue
  2. brain tissue
  3. muscle tissue
  4. liver tissue
  5. tumor tissue
A

5.

remember

Metabolic Quotient of :

early stage embryo ≈►brain tissue

blastocyst is similar to ≈►tumor tissue

48
Q

Artificial oocyte activation is proposed when

  1. A few oocytes failed to fertilise
  2. Less than half the oocytes fail to fertilise
  3. More than half the oocytes fail to fertilise
  4. All the oocytes fail to fertilise
  5. All the mature oocytes fail to fertilise
A

5

49
Q

A Common Ionphore used is

  1. A23185
  2. A23187
  3. A13289
  4. A23191
  5. A23193
A

2.

50
Q

The Common Ionophore Is Known As

  1. Calcimycin
  2. Calcomycin
  3. Calcamycin
  4. Calcmycin
  5. Calcemycin
A

1.

51
Q

Oocyte Activation

A

Reinitiates the cell cycle

Induces oscillations in free calcium levels

Controls the timing of subsequent cell divisions

It is assumed to do so by overcoming a failure to up-regulate free calcium levels after sperm entrance into the Ooplasm

52
Q

The pattern of calcium oscillations (transients)

differs between

normally fertilized and artificial oocyte activation (AOA)

A

true

53
Q

the use of Ionophores in IVF

A

Should be considered experimental

Be restricted to only total fertilisation failure

Has been labelled as experimental by the HFEA

has the potential to be included in all ICSI cases to minimise the risk of failed fertilisation

The HFEA Recommendations:

recommends it use to restricted to selected patients e.g. PLC deficiencies

recommends that patients are provided with its efficacy and potential risks

54
Q

a post hoc indication

A
55
Q

logistic regression to evaluate interdependency

A

logistic regression to evaluate interdependency.
-Evaluating the interdependency between a variety of factors influencing success rates. Here are some specific examples:
-Evaluate the influence of environmental factors like temperature, pH, and oxygen levels on embryo development
-Compare different culture media formulations by using media composition as predictors and embryo developmental
-Analyze the relationship between sperm parameters like motility, morphology, and concentration with fertilization rates or embryo quality using logistic regression.

56
Q

What is the laboratory decisions in case of oocyte with large PB?

A

oocyte with large PB

(this may not be possible for oocytes that are inseminated via IVF, rather than ICSI).

The size of the polar body should only be noted if it is exceptionally large.

It was the consensus that oocytes with an abnormally large polar body should not be inseminated, due to the risk of oocyte aneuploidy.

57
Q

Oocytes with sER disk ?

A

oocytes with sER should not be inseminated. In addition, it was noted that the sibling oocytes should also be examined for the presence of sER disks

58
Q

Large vacuoles

A

Large vacuoles (>14 mm in diameter) are associated with fertilization failure.

In oocytes that are fertilized, those vacuoles that persist past syngamy can interfere with cleavage planes, resulting in a lower blastocyst rate

59
Q

pronuclei with no NPBs called?

A

pronuclei with no NPBs called “ghost pronuclei’

while

a single NPBs (‘bulls-eye pronuclei”)

60
Q

Multinucleation assessment on Day 2 is more reliable than Day 3 ?

A

Multinucleation assessment on Day 2 is more reliable than Day 3

due to difficult assess the small size blastomeres on Day 3 compared to D2

61
Q

early compaction on Day 3 indicates ……?

A

Early compaction on Day 3 is atypical, of unknown biological significance.

62
Q

Final Sperm conc after processing for convential IVF?

A

-After processing by sperm wash or DG,
-Final concentration and motility are reassesed.
Motile sperm are suspended in an amount of media that allows final conc 25*10^3 -50 *10^3 motile sperm per mL.

63
Q

Main components of the microinjection setupMicroscope for (ICSI)

A

ICSI setting includes
an inverted microscope capable of 400-600x magnification connected to a video system.
The microscope has a heated stage, two electric motor-driven coarse- and fine-control manipulators, and two microinjectors.
The microscope must be equipped with Nomarski or other phase interference optics

64
Q

True or false:
PGT-A allowed single ET

A

Euploid embryo after PGT-a is highly recommended for Single ET

65
Q

Epigenetics

A

Changes(reprogramming) in Gene expression by mechanism rather than DNA
-Histon modification
-DNA methylation

66
Q

women 47 old should be encouraged to ART

A

women 47 old should discourage to ART

67
Q

women attende to embryo tansfer but hasband didn’t, and she told that, he already signed the consent yesterday, what’s your decision?

A
  • postpone ET or freez all
  • Inseminatin and ET should be performed in presence of the couple.
68
Q
A

Clinical Embryologist's exam samples. (22 decks)

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