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ASCP CG Certification Practice
What is the best agent to use against mycoplasma? - ✅ BM cyclin - a combination of two antibiotics (tiamulin and minocycline) that effecticely targets mycoplasma growth For successful harvest, each new batch of hypotonic solution should be evaluated by: - ✅ comparing the new lot to a lot already in use Colagenase, pronase, and trypsin are: - ✅ Proteolytic enzymes - break down tissues into single-cell suspensions When harvesting peripheral blood cultures, asceptic technique no longer matters following incubation with: - ✅ Colcemid - mitotic spindle poison that halts cells at metaphase Samples that arrive at the cytogenetics lab should first be: - ✅ Logged in to the system Phytohemagglutinin (PHA) stimulates which type of cells into division? - ✅ T-cell lymphocytes To avoid culture failure, monitoring reagents is crucial. When logging a culture failure, what is the MOST important to document? - ✅ Expiration dates of reagents used Which type of microscope is used when cleaning a CVS (chorionic villus) sample? - ✅ Dissection Which cell type is most problematic if found in a CVS (chorionic villus) sample? - ✅ Decidua (maternal tissue) When looking at a slide under a phase contrast scope and a lot of cytoplasm is still seen around the mets, what is the most likely cause? - ✅ innappropriate slide drying time What will help prevent microbial contamination and cross contamination of cultures? - ✅ Asceptic technique When there are not enough mets to complete the study of a patient/case, what needs to be done next? - ✅ prepare additional slides from any remaining sample When looking at a dropped slide under a phase scope and mets are very spread out and sparse, what could help fix the issue? - ✅ Increading the drying time by using warm slides What is true of an inverted microscope? -
✅ the objectives are beneath the stage High-resolution banding is indicated for a possible diagnosis of: - ✅ DiGenorge syndrome -- microdeletion of 22q11, also called velocardiofacial syndrome "velo" - soft palette, "cardio" - congenital heart defects, "facial" - unique abnormal facial features, short philtrum (upper lip indent), tubular nose, wide spread eyes, etc When verifying if a specimen is appropriate for cytogenetic analysis, what is the MOST important data to accompany the specimen? - ✅ Reason for referral Which type of specimen should be requested for a cytogenetic analysis for turner syndrome? - ✅ Heparinized peripheral blood stored at room temperature (NOT FROZEN) A blood sample can be used to study the cytogenetics of a malignancy only if: - ✅ Dividing leukemic cells are present in circulation All of the cultures in a CO2 incubator failed, what aspects of the incubator are important to monitor to assess the source of the problem? - ✅ Temperature, CO2 level, and Humidity level The most important factor affecting the quality of chromosomes viewed under a phase contrast scope is: - ✅ Slide drying after the cells have been dropped What is the modal number (default number of chromosomes in a cell) for a Down syndrome translocation carrier? - ✅ 45 What is the best tissue to use for both a good success rate and abnormality detection in spontaneous abortion miscarriages? - ✅ Chorionic villi What is a possible cause for a drying oven to not maintain a proper temperature? - ✅ The door does not seal properly If a tech with a cold and sneezing does not take proper precautions while setting up amniotic fluid samples, the specimen could become contaminated with: - ✅ Mycoplasma --> the most common contaminant by a sick tech and can be detected through a specific stain used on the sample
✅ The active regions of the chromosome aka euchromatin -- gene-rich and GC rich DNA, early replicating Which of the following is not assocated with a microdeletion?
- Prader-Willi
- DiGeorge
- Rubinstein-Taybi
- Langer-Giedion - ✅ 3. Rubinstein-Taybi Associated with an autosomal dominant mutation in the CREB or CREBBP gene(s) Rarely assocated with a deletion in 16p13.3 (12% of cases, not the predominant cause, detected by FISH) Presents as facial dysmorphism, broad thumbs, broad big toes, and growth and mental retardation The dark bands produced by routune GTG banding represent: - ✅ AT-rich regions of the chromosome aka heterpchromatin, and gene-poor chromatin, late replicating DNA If GTG banded chromosomes appear ghost-like with little visible banding, one should alter banding prep by: - ✅ Decreasing trypsin time (chewed out and non-distinct) If 50 out of 200 interphase nuclei have 3 signals in a prenatal aneusomy FISH analysis for chromosome 18, the results are: - ✅ Consistent with trisomy 18 or a structural anomaly of chromosome 18 If the same hybridization incubator is used for several different probe strategies, the stringency for each probe can be adjusted by: - ✅ adjusting the formamide and salt concentrations Optimal (best) banding quality is achieved for each patient sample by banding: - ✅ one trial slide per patient to ensure BEST quality In fluorescent microscopy, the exciter and barrier filters used depend on ___? - ✅ Stain - because the wavelength for emission can vary per type of fluorescent stain used (i.e. DAPI) What is a possible reason a metaphase cannot be located when switching from the 40x lens to the 100x oil lens? - ✅ The 100x objective is slightly unscrewed (can occur when cleaning the scope) What is an action a technologist can take to reduce the risk of repetitve motion injuries when doing analysis on a computer? - ✅ Supporting the arm that moves the mouse, Standing and moving around often, and
alternating hands while operating the computer mouse What is the correct modal number for a patient with Turner syndrome with an isochromosome Xq? - ✅ 46 -- because the isochromosome replaces the normal X, causing Turner syndrome What should be used on a cultured CVS sample to obtain more slides for analysis? - ✅ Trypsin-EDTA -- to remove adherent cells from the flask or coverslip to split into more vessels EDTA improves tryp's effectiveness (chelating agent) Cyto studies were ordered on a prenatal patient at 12 weeks gestation after an abnormality was detected via ultrasound. 2 of 15 colonies from seperate primary cultures showed an abnormal cell line of 47,XX,+21. The next step would be to ___? - ✅ Report the result as this is a true mosaic case and suggest: repeating amniocentesis, repeating a detailed ultrasound, or request percutaneous umbilical blood sample (PUBS) After staining, the chromosomes appear to have indistinct bands with little contrast. The problem is corrected by: - ✅ Increasing trypsin exposure (chroms were muddy and dark) When scoring FISH for DiGeorge syndrome, a tech notices the TUPLE1 signal is present, but one of the control ARSA probe signals is missing in every cell. This result suggests __? - ✅ The patient has a deletion of the ARSA region CLIA 1988- (Clinical Laboratory Improvement Amendments) stipulates that equipment must be recalibrated __? - ✅ Every 6 months (Standard 493.1217) What type of stained chromosome would produce a better computer image? - ✅ Lightly stained chromosomes (best contrast and can be digitally edited easier) What is the nomenclature on a patient with down syndrome referred for a hematologic disorder where only trisomy 21 was found? - ✅ 47,XY,+21c[20] 24 hours after culture initiation, a single dish from an amniotic fluid turns cloudy. Which cultures should be disposed? - ✅ The single cloudy dish only What is used to reduce mycoplasma contamination? - ✅ BM cycline CLIA '88 stipulates that equipment must be recalibrated ____? - ✅ After a major repair to ensure still operational How should caustic and corrosive chemicals be stored? - ✅ Both should be stored in a nonflammable cabinet
Chromosome 12q Chromosome 17q The morning after being set-up, a single dish of an amniotic fluid culture turns cloudy and appears contaminated. None of the other cultures set-up that day appears contaminated. What should be done first? - ✅ Harvest the culture as soon as possible When the addition of nonhomologous material to the q-arm of chromosome 16 is suspected after G- banding, what banding technique should be used? - ✅ C-banding What chromosome is most different between C- and G-banding? - ✅ Chromosome 9 What chromosome region does quinacrine and acridine orange dyes stain? - ✅ AT rich DNA Under an electron microscope, what is observed as a consequence of all banding procedures? - ✅ Chromosomal collapse Case #777 has an uncertain diagnosis after G and C bands. The director suspects satellites are involved with the chrom marker in question. What banding/stain should be used to confirm satellites are in question? - ✅ NOR banding What procedure should be performed first if sequential banding is required? - ✅ Q-banding A tech suspects an inversion of chrom 9 with G-banding analysis. What banding technique would confirm this? - ✅ C-banding What chromosome polymorphisms can be detected with Q-banding? - ✅ Y-chrom polymorphisms Chrom 3 polymorphisms Poly's of the satellites of acrocentrics After G-banded analysis, a cyto director suspects chrom 10 has a terminal del. What banding procedure would be MOST helpful in determining if a deletion has occurred? - ✅ FISH Cytogenetic studies have been requested for a patient with a diagnosis of questionable leukemia. This patient has a low white cell count, circulating myeloblasts, and has received chemotherapy and a blood transfusion. What procedure is LEAST likely to be informative? - ✅ PHA-stimulated blood culture Special culture conditions or medium are required to demonstrate what conditions? - ✅ Fragile X - folic acid deficient medium
Inactive X - use bisulfite, but now diagnosed via molecular methods Sister chromatid exchange - BrdU culture medium 46,XY,der(9)(t(9;10)(q32;q12). What term best describes this written karyotype? - ✅ Unbalanced ("der" means "a derivative chromosome"; "t" translocation. This indi. has only one copy of chromosome 9q from band 9q33 to 9qter), therefore is unbalanced What is the CORRECT ISCN nomenclature for a male with fragile X syndrome? - ✅ 46,Y,fra(X)(q27.3) (the normal sex chromosome is written first followed by the abnormal sex chromosome) NOT 46,XY,fra(X) bc that indicated the presence of another X (= 47 chroms) The peripheral blood culture of a patient suspected of having CML is 46,XY. In order to rule out the presence of the t(9;22) the following tissue would be cultured next: - ✅ Bone marrow (sometimes, the cancer cells go into circulation blood. The bone marrow will be the origin of the cancer cells) Twenty-nine out of 30 metaphases are normal in the amniotic fluid culture of a pregnant woman who has a history of multiple miscarriages. One cell has a translocation. What does this signify? - ✅ Cultural artifact (in order to be considered as mosaicism, you have to have more than one cell in 30-cell count, and it needs to be confirmed by skin tissue if possible) A Cri-du-chat patient has a chromosome abnormality of: - ✅ r(5)(p15.1q35) (this ring chromosome is missing the tip end of 5p and the tip end of 5q. Deletion of 5p15 is cri-du- chat syndrome) When an abnormal cell is detected in an in situ culture all of the following should be considered to rule out pseudomosaicism: - ✅ Confirm the abnormality in other cells in the colony. Confirm the abnormality in another culture. Confirm the abnormality in other colonies on the same coverslip. The optimal objective lens for capturing chromosome images is ___. - ✅ Planapochromatic (this lens is corrected for the both color and spherical aboration) If an incubator runs out of carbon dioxide for 6 hrs, what would you expect? -
One should not use the aperture diaphragm to: - ✅ Control brightness of light For the imaging of chromosomes, I want the best resolution possible, so I always use: - ✅ An obj lens with an NA of 1.3 to 1. Koehler illumination should be performed daily to observe chroms under the scope. How does one describe the process of koehler illumination? - ✅ The process of centering the light source and spreading the light uniformly over the field of view The light histogram on the monitors of an automated karyotype system is used to: - ✅ Achieve optimal light intensity for the met being captured, set the threshold which affects the contrast (A and B values), and to enhance the image selecting values above or below which a pixel is either white or black What type of objective provides the greatest depth of field and field of view? - ✅ Low power After performing koehler illumination, what other scope part should be adjusted to ensure optimal contrast between light and dark bands of the chroms? - ✅ The condenser diaphragm should be closed down by 1/4 to 1/ The primary limiting factor for resolution in a light microscope is: - ✅ The wavelength of light r = (wavelength)/2NA NA can be adjusted through the type of objective being used, so the only part that cannot be actively changed is the wavelenth of light Why is it necessary to adjust the amount of light after changing the objective lens? - ✅ Higher magnification needs more light, Proper lighting is needed to see specimen detail, and the lens aperture decreases with higher magnificaction lenses When reviewing a final karyotype a possible overlapper chromosome is suspected. What is needed to confirm the chrom # of this cell? - ✅ ID # of the patient, Scope used to capture the cell analyzed, and the coordinates of the met A fetus with an increased risk for trisomy 18 would have a serum analytes that is: - ✅ low AFP, low hCG, and low uE AFP = Alpha feto protein hCG = human chorionic gonadotropin uE3 = Unconjugated estriol Parental chrom analysis would be recommended for these karyotypic abn's: -
✅ 46,XX,der(14;21)(q10;q10),+ del(13q) dup(11q) 50% of cells from an amniotic flask have 92,XXYY and the other 50% have 46,XY. This probably indicated: - ✅ Pseudomosaicism The type of cell analyzed with the CVS direct preparation is: - ✅ cytotrophoblast Peripheral blood G-band karyotype analysis is requested for all the following: - ✅ Multiple malformations seen in a newborn Mental retardation of unknown etiology Multiple miscarriages A technologist wants to stain chromosomes using a banding technique that manifests AT-rich regions on chromosomes. What staining procedure should he use? - ✅ QFQ What is the optimum pH for the trypsin? - ✅ 7.2 - 7. Quinacrine molecules bind to the DNA by: - ✅ Binding to the AT rich regions What statements about G-banding are true? - ✅ Methanol-acetic fixation is necessary for chromosome spreading and has aneffect on G- banding, G-dark bands are late replicating DNA regions, G-banding correlates with the chromomere pattern of pachytene chromosomes of meiosis. Which location can be shown by C-band? - ✅ Constitutive heterochromatin The culture method used to obtain chromosome preparations from amniotic fluid is considered: - ✅ Short term culture What is the purpose of establishing a PHA-stimulated blood culture for a leukemia patient? - ✅ To see if an abnormality is constitutional rather than acquired What is the purpose of establishing an unstimulated blood culture for a leukemia patient? - ✅ To check for circulating immature cells in the peripheral blood. All of the following ARE characteristics of the horizontal laminar flow hood? - ✅ Protects the culture from contamination Should not be used with transformed cell lines. Sterlizes the air entering the hood
What happens to the enzymatic activity of trypsin as pH increases to 8? - ✅ The enzymatic activity increases What do you add in order to detect SCE (sister chromatid exchange)? - ✅ BrdU Media in the incubator has turned magenta and cell death has occurred. Most likely the: - ✅ CO2 was too low You have just received fetal and placental tissue suspected of fungal contamination. How do you culture this sample? - ✅ Fetal tissue w/ Nystatin What is true about slide preparation for FISH? - ✅ Slides should be kept at room conditions. When using the Applied Imaging system to capture the FISH image, which program should be chosen to use? - ✅ Probe The M-FISH procedure makes use of combinatorial labeling in order to uniquely label all chromosomes human chromosomes. Which of the following represents the correct mathematical equation for determining the number of combinations based on the number of fluorochromes (n) available? - ✅ (2^n - 1) (i.e.) How many combos of 3 fluorocromes are there? (2^3 -1) = 7 What is the maximum number of color combinations that can be obtained from 5 fluorochromes? - ✅ 31 Which of the following ARE advantages of FISH diagnostics concerning interphase testing? - ✅ Can rapidly detect sex chromosomes or aneuploidy of chromosomes. Requires no culture Requires only simple microscope analysis. The temperature of -20oC is necessary in which of the following FISH procedures? - ✅ Probe storage The temperature of 4oC is necessary in which of the following FISH procedures? - ✅ FITC-avidin storage The temperature of 90oC is necessary in which of the following FISH procedures? - ✅ simultaneous denaturation Most important pretreatment to age slides for FISH is: - ✅ 2x SSC at 37 degrees for 30 minutes What would you do if you see high non-specific background on the FISH slide just postwashed? -
✅ Increase postwash temperature Which is preferred as a metaphase probe? - ✅ SKY probe A MLL FISH gave the following signal pattern 1F 1R 1G. What is the ISCN of this cell? - ✅ nuc ish (MLLx2)(5?MLL sep 3?MLLx1) Fluorescence can be caused by what light source? - ✅ Mercury vapor lamps To AVOID empty magnification, the total magnification seen by the eye should: - ✅ Not exceed 1000X the N.A. of the objective What ARE functions of the CCD camera? - ✅ Capturing light Controlling the resolution of the image Determination of matrix size based on the number of photo cells How many bands are on chromosome 10 in the 350 to 400 band level range? - ✅ 12 to 14 How many bands are on chromosome 10 in the 425 to 475 band level range? - ✅ 15 to 21 How many bands are on chromosome 10 in the 500 to 550 band level range? - ✅ 22 to 28 AML M0 type - ✅ AML with minimal differentiation (no chromosome change associated) AML M1 type - ✅ AML w/o differentiation (no associated chrom changes either) AML M2 type - ✅ AML with differentiation t(8;21)(q22;q22) AML M3 type - ✅ Acute Promyelocytic Leukemia (APL) t(15;17)(q22;q11-12) AML M4 type - ✅ Acute Myelomonocytic Leukemia (AMML) inv(16)(p13q22) AML M5 type - ✅ Acute Monocytic Leukemia t(9;11)(p22;q23) other 11q23 rearrangements
