Test Questions Flashcards
The following enzymes can be used to dissociate Chorionic villus samples (cvs) except for:
RNAse(correct)
Collagenase
Trypsin
Protease
All of the following have buffering capability except:
Sodium bicarbonate
L-glutamine(correct)
Hepes
FCS
What type of media is necessary for the culture of fragile x cases?
Media with extra FCS
Folic acid-deficient media(correct)
Complete media
Lysine-rich media
The most important reason why biological safety cabinets are used instead of horizontal laminar flow hoods in the handling of human tissues is
To provide a clean working area
To prevent contamination of the specimen
To protect the technologist who is processing the specimen(correct)
To prevent cross-contamination between two different cell lines
In which of the following situations would one suspect bacterial contamination?
The culture media is red-orange
The culture had a dark mass floating on the surface of the media
The culture media is turbid in appearance(correct)
The ph of the media is 7.2
Which of the following can be used for serial culture from amniotic fluid?
Erythrocyte
Lymphocyte
Epithelial cells
Fibroblast(correct)
Which of the following chemicals is used for cryopreservation?
Methanol
Ethyl alcohol
Dimethyl sulfoxide DMSO(correct)
Sodium hypochlorite
Adding the drug Colcemid to cell cultures can cause all except:
Chromosome condensation
Increase in the number of metaphases
Cell swelling( correct)
depolymerization of the spindle apparatus
Which of the following is not a necessary ingredient which is added to modified media
L-glutamine
Vitamins
Antibiotics (correct)
Buffer
During slidemaking all of the following factors are important except for:
Drying rate(evaporation)
Quality of the glass slides
Slides are first dipped in water
Slides are first dipped in acetic acid(correct)
A good bonding pattern will be expected when we observe chromosomes before banding under the phase scope with the following appearance:
Dark(correct)
Pale
Shiny
Separated chromatids
Which of the following factors are not important for chromosome spreading?
Hypotonic treatment time
Dropping the cell from an increased height
Cell suspension concentration
Mitotic index(correct)
If short chromosomes result after harvesting, what change may alleviate this condition for the next harvest?
Increase concentration on methanol in fixative
Decrease exposure to hypo
Decrease concentration of colcemid(correct)
Increase concentration of colcemid
Which of the following apply to mitotic arrestants?
Result in depolymerization of the mitotic spindle
Colchicine
Colcemid
All of the above(+)
Which of the following apply to hypo solutions?
Are responsible for lysing rbc
Cause cells to swell
0.075 M KCL
All of the above(+)
Which statements regarding the cvs procedure are true?
It was introduced in 1968
It involves removal of a sample of placental tissue by transcervical aspiration with a thin plastic catheter
It involves removal of a sample of placental tissue by trans abdominal aspiration with a 20 gauge needle
All of the above(+)
B and C
What prenatal test can be done with the highest accuracy and as early in the pregnancy as possible?
Percutaneous umbilical cord blood
CVS(+)
Amniocentesis
Maternal serum triple screening
Which of the following has the lowest toxicity to exposed cells?
Sodium heparin(+)
Lithium heparin
Sodium hydroxide
Gentamicin
A pathologist is sending solid tumor specimens to a lab for analysis. What instructions should be given to the pathologist concerning the preparation and transport of the specimen to ensure the highest chance of successful culture?
Ensure the sterility of the specimen by placing it in formalin until it reaches the lab
Ensure that the specimen does not contain necrotic or fatty tissue(+)
Ensure that the specimen is transported on dry ice to prevent decomposition
All of the above
It is possible for the collecting syringe used in amniocentesis to be toxic to the amniotic cells
True(+)
False
A solid tumor specimen cannot be shipped from the hospital to a lab in another city due to a major holiday. How should the sample be stored so that the cells will remain alive until they can be delivered and placed in culture?
The specimen should be preserved by placing it in formalin until it reaches the lab
The specimen should be placed in media and then in a 37C incubator until it can be shipped
The specimen should be stored at -20C or on dry ice to prevent cellular decay
The specimen should be placed in media and then refrigerated until it can be shipped(+)
Blood is received for cytogenetic analysis in a busy clinical laboratory. It will be several hours before a cytogenetic technologist will be able to do the set-up procedure. How should this blood be stored during the interim period?
The blood should be stored at room temp(+)
The blood should be stored on ice
The blood should be stored in the freezer at 0C
All of the above
What is optimal temp for transport of amniotic fluid specimens?
37C
40C
0C
20C(+)
All of the following will result in culture failure except:
Transport at room temp(+)
Sample shipped on ice
Specimen exposed to 40C
Improper sterile techniques used
When amniotic fluid specimens are collected for culture, which of the following are safeguards against culture failure?
Samples are collected in one syringe
The first 5cc of fluid are discarded
Two cultures are set up with two different types of media(+)
T12 1/2 flasks are used
Of the tissue types selected from abortus specimens, the one that exhibits the best growth rate is:
Fetal skin
Spleen
Chorionic villi(+)
Umbilical cord
The type of cell analyzed with the CVS direct preparation is:
Yolk sac endoderm
Cytotrophoblast(+)
Mesenchymal stroma
Syncytiotrophoblast
When analyzing lymphomas for cytogenetics, lymph node biopsies are the tissue of choice
True(+)
False
Determining the wbc in bone marrow is important because:
Patients who have a low wbc have a worse prognosis
It is important to ensure that all cultures have the same number of dividing cells
It is important to maintain uniform culture conditions, and cell concentration varies greatly from sample to sample(+)
BM cells will not grow in culture unless the cell density is at least one million cells per ml
What is the recommended final cell concentration of cells for BM cultures for chromosome studies?
1mln cells per ml of media(+)
2-3 —//—
3-4 —//—
5 —//—
Solid tumor culture failure can be caused by which of the following?
Specimen contamination
Specimen necrosis
Growth of normal fibroblast cells
All of the above(+)
When a clotted BM specimen comes into the lab, the technologist should:
Request a new specimen
Discard the specimen
Place the specimen in a warm waterbath and attempt to break up clot(+)
Set up the clotted BM as usual
Peripheral blood G-band karyotype analysis is requested for all the following except:
Multiple malformations seen in a newborn
Single malformation seen in a newborn(+)?corr
Mental retardation of unknown etiology(+)?
Multiple miscarriages
Which of the following is not an indication for routine peripheral blood chromosome analysis?
Advanced maternal age(+)
To determine whether a chromosomal abnormality is constitutional or acquired in cancer patients
To study chromosome damage due to environmental exposure to chemicals, drugs, radiation
Multiple congenital anomalies with mental retardation
Failure rates for cytogenetic analysis of BM specimens can be influenced by:
Recent treatment of the patient The specific disease Amount of specimen Time between collection of specimen and set up of culture All of the above(+)
Which of the following samples would you assign the highest priority?
A peripheral blood sample from a 24 yo woman who has had three miscarriages and one normal child
Amniotic fluid from a 36 yo woman who is 18 wks pregnant
A body who is in the ICU with multiple malformations(+)
A13 yo child with ambiguous genitalia, moderate mental retardation and developmental delay
Which of the following situations would be the most likely to benefit from high resolution banding?
Detecting a complete deletion of the short arm of chromosome 10
Detecting an i(8q)
Detecting the extra Y in a 47,XXY case
Detecting a microdeletion on chromosome 15q(+)
2 ml of 20x10^6 cells/ml BM was received. How many 10 ml cultures of 1x10^6 cells/ml can be set up?
1
2
3
4(+)
A tissue sample has been received in isotonic solution after being in transit for 3 days. What should you do first?
Set up the sample immediately
Rinse in transport media with antibiotics(+)
Reject the sample, since it has been in transit for too long time
Rinse in fresh isotonic solution
Which of the following is most likely to result in culture failure?
Collection in a non-sterile container
Fungal contamination
Collection of sample in EDTA(+)
Receiving sample after 2 days
When dropping slides, the majority of chromosomes appear to be short. This can result from which of the following?
A long exposure to Colcemid(+)
A short exposure to Colcemid
A short exposure to hypo
Improper fixation
If short chromosomes result after harvesting, what change may alleviate this condition for the next harvest?
Increase concentration of methanol in fix
Decrease exposure to hypo
Decrease concentration of colcemid (+)
Increase concentration of colcemid
After what period of time do PHA peripheral blood lymphocytes reach their first division?
12 hrs
24 hrs
36 hrs
48 hrs(+)
Which of the following is information that is important to consider when freezing cells?
Age of the culture
Concentration of cells(+)
Cell type
Media used
24 BM and 72 BM LPS cultures will be set up on a new CLL with 50*10^6WBC
True(+)
False
Which of the following may cause cell death?
5%CO2 atmosphere
A 24 hr delay in culturing
A ph of 7.2
Incubation at temperature of 40C(+)
For optimal cell growth, which of the following ph will provide that?
- 8-7.0
- 0-7.2
- 2-7.4(+)
- 4-7.6
Which of the following tissues grows most slowly in culture?
Fetal tissue
Skin(+)
Gonad
Amniotic fluid cells
Which of the following media supplements is optional for long-term cultures?
L-glutamine
Antibiotics antifungicides(+)
Growth factors
Fetal calf serum
Which of the following best describes a media that has a low serum level?
Defined
Maintenance(+)
Growth
Complete
The statements below refer to culturing methods. Which statement is False?
The preferred mode for cytogenetic analysis of solid tumors is long-term culture(+)
The success rate of cytogenetic analysis of solid tumors is less than that of leukemias currently.
Culture of epithelial tumors is more difficult than other types of tumors
A Biosafety cabinet is required for tissue culture
Regarding the following statements, which is False?
Epithelial cells cannot be morphologically distinguished from mesenchymal cells in culture when viewed under an inverted phase contrast microscope(+)
Contamination with normal fibroblasts can give an erroneous result
Different media are used in the culture of different tumor types
All of the above
Harvest of a 14-day-old amniotic fluid culture results in many cells, but few mitoses. The most likely cause of the low mitotic rate is:
Senility of the culture
Chromosome abnormality
Maternal age
Contact inhibition due to high cell confluence(+)
The culture method used to obtain chromosome preparations from amniotic fluid is considered:
Short term culture(+)
Intermediate term culture
Long term
Non of the above
The fixative step of cell harvesting does not:
Kill the cell by fixing the cell membrane
Remove water from the cells
Extract nuclear histones
Disperse the chromosomes (+)
Which of the following is not a mitotic inhibitor
Colcemid
Vinblastine
Ethidium bromide(+)
Colchicine
Carnoy’s fixative is the standard fixative used in chromosome harvesting procedures. Which of the following is a description of this solution?
3(glacial acetic acid):1(methanol)
1(glacial acetic acid):2(methanol)
2(glacial acetic acid):1(methanol)
1(glacial acetic acid):3(methanol)(+)
If the metaphase portion of the cell cycle is 15 min and a cytogeneticist chooses to treat a peripheral blood specimen with Colcemid for 45 min, what is the increase in mitotic cells, which will be observed after the harvest procedure is completed?
100%
200%
300%(+)
400%
What is the concentration of KCL most often used for harvesting?
- 04 M
- 06 M
- 075 M(+)
- 75M
How many grams of KCL is added to 1000 ml of double deionized water to make 0.075M KCL (m.w=74.4)?
- 58g(+)
- 79g
- 79g
- 5g
Mitotic arrestants prevent cells from entering:
S phase
Prophase
Anaphase(+)
Telophase
A technologist is having a problem with cytoplasm remaining around the cells. Two extra fixative steps have been added, but the problem persists and the cell pellet is white and fluffy in appearance. What else can this technologist do to alleviate this problem?
Manipulate the drying time(+)
Decrease the methanol concentration
Wash 2 more times with fix
Drop the cell suspension onto the slide from a lesser height
Which of the following factors concerning hypo solutions do not affect chromosome spreading
Concentration of the hypo solution
Temperature of the hypo solution
Time period cells are exposed to the hypo
Number of washes with hypo(+)
Disadvantages of the direct harvest method as opposed to harvesting cultured cells for CVS analysis are:
Poor quality chromosomes and increased pseudomosaicism(+)
Increased incidence of maternal cell contamination
Requires fluorescent marker studies
Increased pseudomosaicism
Cytogenetic studies have been requested for a patient with a diagnosis of questionable leukemia. This patient has a low white count, circulating blasts, and has received chemotherapy and a blood transfusion. Which of the following procedures is least likely to be informative?
Direct harvest of BM
Unstimulated culture of peripheral blood lymphocytes
PHA stimulated blood culture(+)
Unstimulated culture of washed blood cells
PHA is a common additive to blood cultures. What category of additive is it?
A mitogen(+)
An anticoagulant
Growth serum
Arresting solution
On Monday morning the tech noticed widespread cell death in incubator A. He checked the environment and conditions of the cultures. What condition would likely be the cause of the cell death?
5% CO2 atmosphere
40C incubator temperature(+)
Ph of 7.3
All conditions would cause cell death
The following are indicators for amniocentesis except for:
Maternal age
Parental chromosome abnormality
A family history of an X-linked disorder
A family history of an autosomal recessive disorder(+)
How long do you wait before requesting an amniotic fluid if no cell attachment is observed?
3days
10days(+)
21days
28days
Which media component is unstable?
serum
L-glutamine(+)
antibiotics
amino acids
What is the purpose of establishing a PHA-stimulated blood culture for a leukemia patient?
To check for circulating immature cells in the peripheral blood.
To see if an abnormality is constitutional rather than acquired(+)
To look for early rearrangements which may have predisposed this individual to develop cancer.
To study leukemic cells of T-cell origin
What is the purpose of establishing an unstimulated blood culture for a leukemia patient?
To check for circulating immature cells in the peripheral blood(+)
To see if an abnormality is constitutional rather than acquired.
To look for early rearrangements which may have predisposed this individual to develop cancer
all of the above
PHA will affect which of the following types of cells?
B-cells
Platelets
T-cells(+)
Phagocytes
This type of culture has a decreased turn around time and allows easier distinction between mosaicism and pseudomosaicism
Primary amniotic fluid cell culture
In situ-amniotic fluid culture(+)
Flask-amniotic fluid culture
Short term amniotic fluid culture
The lab supervisor detected contamination in a 5 day old in situ amniotic fluid culture. This was one of four
cultures set-up on this patient. What would be the best course of action in this situation?
Increase antibiotic concentration
Refeed the culture
Transfer the specimen to another culture vessel
Harvest immediately(+)
How would a technologist know when to harvest an in situ amniotic fluid culture?
By the number of days in culture
Within 12 hours of subculture
The number of colonies present(+)
Two of these
The major difference between the in situ and flask methods lies in the harvest technique.
True (+)
False
Monday morning around 2:30, Incubator A ran out of carbon dioxide. What change in the cultures, if any would
you expect for the technologists to find at 8:00 am Monday.
About the same pH
A lower pH
A higher pH(+)
A moderate pH
Culture reagents and chemicals may be used without adverse effects after they have expired
True
False(+)
Amniotic fluid cultures are generally started in a medium with what percentage of serum.
5%
10%(+)
15%
20%
PHA
Stimulates the large lymphocytes
Transforms polymorphonuclear leukocytes
Causes decrease in RNA activity
Causes peak in DNA synthesis at 72 hours(+)
Is needed throughout cell cycle to produce sufficient metaphases
PHA
Swells cells Removes water from cells Stimulates mitotic activity(+) Precipitates nucleic acid Prevents clotting
In short term culture the following is/are true:
Incubation in a CO2 atmosphere is essential.
Most commercially prepared media is suitable(+)
Plasticware is never toxic to cells.
All of the above.
“Starring-in” is a method of
Obtaining the specimen from the patient Mincing the tissue Immobilizing the tissue in the culture vessel(+) Fixing the cells Preparing the slides
A change of 1 pH unit in culture medium is caused by a ________ change in CO2 concentration.
10-fold(+) 25-fold 50-fold 100-fold 1000-fold
Five percent CO2 results in a pH of
- 5
- 8
- 0
- 4(+)
- 8
What is the most commonly used growth supplement for culture medium?
Insulin
Transferrin
L-glutamine
Fetal bovine serum(+)
When subculturing a long-term cell line, a technician finds the cells are not easily removed by their trypsin
solution. What can be added to the trypsin to increase its effectiveness in removing the cells from the flask?
Serum
Calcium and magnesium
EDTA(+)
L-glutamine
Which of the following microorganisms can adversely effect long-term cultures?
Mycoplasma
Bacteria
Fungus
All of the above(+)
Which of the following is NOT an advantage of tissue culture (in vitro) studies over patient studies (in vivo)?
Less expensive
More ethical
Possibility of contamination(+)
Can directly target cells with test agent
What microscope is used to view living monolayer cell cultures?
Brightfield microscope
Fluorescent microscope
Stereomicroscope
Inverted phase microscope(+)
Unstimulated blood cultures grow out of what type of cells?
Segmented neutrophils
Lymphocytes
Blast forms(+)
Monocytes
Stimulated blood cultures grow out what type of cells?
Basophils
Lymphocytes(+)
Immature forms
What is NOT a principle behind the culturing of solid tumors?
Culturing of living cells provides better morphology chromosomes for analysis than chromosomes obtained by direct harvest.
To generate an expandable cell population which may be used for sequential and long-term experimentation.
To determine which chromosomal rearrangements are due to in vitro artifacts vs. those which represent the true in vivo condition.(+)
None of the above are principles behind the culturing of solid tumors.
Collagenase is used in the analysis of solid tumors to:
Dissociate cells(+)
To supplement culture media
To prevent mycoplasma contamination
To subculture the monolayer cultures.
The inverted microscope is essential in the monitoring of cell cultures by allowing the technologist to visualize all but which of the following cellular features?
Cellular confluency
Chromosomal abnormalities(+)
Mitotic cells
Cellular attachment
The next day after a solid tumor specimen has been set up in culture the tissue culturist notices that the culture medium has turned a bright yellow color. When the flask is observed under the inverted microscope, a large number of small dot shaped structures are observed. What is the most likely cause of this pH change and the appearance of these structures?
The specimen was received from the necrotic center of the tumor.
The specimen was not collected under sterile conditions.(+)
The specimen was not grown in the appropriate media.
The specimen was not transported at the appropriate temperature.
Which of the following is not an effect of fixation?
Remove water from the cells
A short exposure to Colcemid(+)
A short exposure to hypotonic
Improper fixation
Upon what cell structure does Colchicine have its effect?
Centrioles
Spindle fibers(+)
Centromeres
Chromosomes
This chemical is more toxic than its more recent synthetic counterpart.
Thymidine
KCL
Colchicine(+)
Ethidium bromide
This chemical is used to synchronize cells.
Thymidine(+)
KCL
Colchicine
Ethidium bromide
Which of the following is not a mitotic stimulant?
PHA
Citrate(+)
Pokeweed
LPS
Carnoy’s fixative:
Swells cells
Stops hypotonic action(+)
Stimulates mitotic activity
Prevents clogging
Which of the following chemicals is NOT an intercalating agent?
Actinomycin D
Ethidium bromide
Methotrexate (Amethopterin)(+)
Quinacrine dihydrochloride
This cell cycle blocking agent causes cells to be arrested in the S-phase of the cell cycle through the inhibition of dihydrofolate reductase.
Methotrexate (Amethopterin)(+)
5-Bromodeoxyuridine
Ethidium bromide
Bleomycin
This chemical is a thymidine analog that replaces thymidine easily in mammalian cells.
5-Fluorodeoxyuridine
Adenine
5-Bromodeoxyuridine(+)
Hoechst 33258
Which of the following environmental factors can have an impact on drying time and therefore impact
chromosome spreading during slide making?
Temperature
Humidity
Air currents
All of the above(+)
A new hypotonic solution causes overspreading of chromosomes. What technique can be used to prevent the overspreading?
Drop cells on a wet slide.
Increase the height from which cells are dropped.
Decrease the cell concentration
Reduce the drying time(+)
The hypotonic treatment of cells during the harvesting procedure is used to:
Remove water from the cells
Disperse the chromosomes(+)
Enhance chromosome morphology
Remove the H1 fraction of histones
All of the following may be used for a hypotonic during the harvest procedure EXCEPT:
Methanol(+)
Sodium citrate
Water
Potassium chloride
You have 2 ml of peripheral blood. What sort of cultures should you set-up with this blood?
One 24 hour culture
One 24 hour and one 48 hour culture
Two 72 hour cultures(+)
One 120 hour culture
The cell cycle can be blocked by using which of the following:
Ethidium bromide
Excess thymidine(+)
BrdU
Mitomycin-C
All will result in reduced mitotic index, EXCEPT:
Not adding PHA-m to culture media before culturing
Adding twice the amount of Colcemid(+)
Allowing chromosomes to overspread on the slides
Treated with hypnotic solution for too long time
A culture that was left in the incubator has turned yellow. What is most likely the cause of this?
Increase in temperature
Decrease in temperature
Increase in pH
Decrease in pH(+)
A tumor in saline arrives in the lab, what do you do?
Add more saline to the tumor
Rinse tumor in culture media and antibiotics(+)
Harvest tumor immediately
Reject the specimen since the tumor should be
transported in transfer media
What is the best time to freeze cells for long term storage?
10% confluent log phase
20% confluent log phase
80% confluent log phase(+)
100% confluent log phase
What happens to the enzymatic activity of trypsin as pH increases to 8?
The enzymatic activity stays the same
The enzymatic activity decreases
The enzymatic activity increases(+)
The enzymatic activity stops
Which of following does not have buffer capabilities?
L-Glutamine(+)
Hepes
Na2HPO4
NaHCO3
Which has the highest chance of Maternal Cell contamination?
Peripheral blood
Amniocentesis
Unidentified POC(+)
PUBS
What do you add in order to detect SCE?
Ethidium bromide
Thymidine
MTX
Brdu(+)
Media in the incubator has turned magenta and cell death has occurred. Most likely the:
Temperature was too high
Temperature was too low
CO2 was too low(+)
CO2 was too high
You have just received fetal and placental tissue suspected of fungal contamination. How do you culture this sample?
Fetal tissue w/ Kanomycin
Fetal tissue w/ Nystatin(+)
Placental Tissue w/ Mycostatin
Placental Tissue w/ Streptomycin
