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CC 4530 Basic Laboratory Techniques > Blood Culture Setup / Harvesting > Flashcards

Blood Culture Setup / Harvesting Flashcards

1
Q

Dividing cells could be stopped at the cell cycle in which phase for chromosomes?

A

M-Phase

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2
Q

Which mitogen does cytogenetics uses?

A

PHA

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3
Q

As a mitogen what does PHA stimulate?

A

T-cells

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4
Q

Between how many hours can peripheral blood cultures stimulated with PHA can be used for chromosome preparations?

A

48-96 Hours

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5
Q

Two tubes of samples were received, one in EDTA and one in sodium heparin; where does each one belong?

A

The sodium heparin tube goes to cytogenetics and the EDTA tube goes to the molecular lab.

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6
Q

What does not have a buffer capacity?

A

L-Glutamine

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7
Q

A magenta color of media is an indication of what?

A

That it is Basic

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8
Q

Combined acetic acid and methanol can do what?

A

Fix/harden the cell membrane

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9
Q

A hypotonic solution can do what?

A

Swell cells

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10
Q

The results of hypotonic treatment depend on all of the following:

A

Composition
Degree
Length of exposure
Temperature

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11
Q

What is excluded, or does not affect hypotonic treatment/chromosome spreading?

A

The amount of hypotonic/the number of washes with hypotonic.

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12
Q

True or False? Cells should be left in the first full fix for some time to allow them to harden before additional fixative washes.

A

True

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13
Q

True or False? Cold fixative will lead to poorer chromosome morphology.

A

False, cold fixative can lead to better morphology.

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14
Q

True or False? Fixative can be stored for days or weeks at room temperature before being used.

A

False, fixative should always be made fresh.

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15
Q

What is the proper ration of Carnoy’s fixative?

A

1:3, acetic acid:methanol

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16
Q

Which step of the harvest procedure makes cells more resistant to damage from centrifugation?

A

Addition of fixative to the hypotonic

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17
Q

Which of the following biological safety cabinets are recommended for use in cytogenetic laboratories?

A

Biological Safety Cabinet Class II

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18
Q

Mitotic arrestants prevents cells from entering what phase?

A

Anaphase

19
Q

What is the concentration of KCL (hypotonic solution) most often used for harvesting?

A

0.075 KCL

20
Q

What stops hypotonic action?

A

Carnoy’s Fixative

21
Q

What are the 4 major steps of blood harvesting?

A

Adding Colsamid
Adding Hypotonic Solution
Adding Pre-Fix
3 cycles of Fixative

22
Q

What temperature is the incubator set for blood culture incubation?

A

37C

23
Q

How many ml of blood and medium do you use to create a culture?

A

9 ml of PB-Max medium to 1 ml of blood specimen.

24
Q

In cytogenetics the anticoagulant of choice for peripheral blood is?

A

Sodium heparin

25
Q

When dropping slides the majority of chromosomes appear to be short. What is the possible reason?

A

A long exposure to Colcemid

26
Q

What supplement in culture media is NOT stable and needs to be added into the media freshly before the culture?

A

L-glutamine

27
Q

The most commonly used PHA in cytogenetics?

A

PHA-M

28
Q

What pH range is optimal for cell growth?

A

7.0-7.4

29
Q

True or False? When doing hypotonic treatment, the optimal time should be the same with different cell types and tissue.

A

False

30
Q

Peripheral blood is used for determining what kind of karyotype?

A

Constitutional

31
Q

True or False? Peripheral blood is used for the determination of the acquired karyotype of patients with hematologic malignancies or solid tumors for comparison abnormalities in their neoplastic cells.

A

False, it’s constitutional karyotypes.

32
Q

What mitogen is used to stimulate B-cells?

A

LPS

33
Q

What kind of system is used for peripheral blood and bone marrow cultures?

A

Closed system

34
Q

What is the difference between open and closed culture systems?

A

Open culture system has vessels that remain open to the environment, closed culture system culture vessels that are tightly closed so there is no free gas exchange.

35
Q

What are the steps to unpacking a biological sample?

A

Always wear PPE.
Use the safety cabinet.
Identify the specimen type and reason for referral.
Document everything, including suspect conditions.
Always wash your hands before leaving the lab.

36
Q

If 0.75M KCL is used in harvesting instead of 0.075M, what will happen?

A

The cell will shrivel up.

37
Q

After a successful harvesting, what color of cell pellet should be?

A

White.

38
Q

What is the function of the fixative solution?

A
  • Kill the cells (precipitates the free hemoglobin)
  • Fixes the cell membrane
  • Removes the H1 fraction of the histone proteins from the chromosomes
39
Q

Which white cells are used for cytogenetic analysis?

A

Lymphocytes

  • T cells
  • B cells
40
Q

What buffer is used in cytogenetics?

A

Sodium bicarbonate

41
Q

Why is a Class II Biological Safety Cabinet used in cytogenetics?

A

Because it protects both the specimen and

worker from contamination.

42
Q

What are the major steps to a blood culture setup?

A
  • Use a sterile pipet to transfer 9 ml of culture, and 1 ml of sample, into a 25 ml culture flask.
  • Close cap tightly and mix.
  • Put the culture flask in 37C incubator, incubate for 72 hours.
43
Q

What is the purpose of harvesting?

A
  • Stop the dividing cells at metaphase stage
  • Get rid of red cell and only keep white cells
  • Prepare for chromosome spreading and banding
44
Q

What is Colcemid?

A

Mitotic inhibitor, stops the cells in metaphase by disrupting the spindle fibers.

CC 4530 Basic Laboratory Techniques (3 decks)

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