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Top 100+ Flow Cytometry Interview Questions And Answers - May 30, 2020

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Top 100+ Flow Cytometry Interview Questions And Answers

Question 1. Flow Cytometry Is A Measure Of Two Things, What Are They?

Answer :

Light scattered via cells and fluorescence from fluorochromes attached to cells.

Question 2. In Flow Cytometry Are You Measuring In All Blood Cells?

Answer :

No, you lyse the RBC's prior to the procedure.

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Question three. What Is Flow Cytometry Used Commonly For?

Answer :

Identify Ag expression styles on cells. Determine if cells are normal or bizarre based on diagnosed styles.

Question four. What Is The Technique Of Flow Cytometry?

Answer :

Fluorescence-labeled Abs incubated with cells. Laser excites fluorochromes (on Abs), which emits fluorescence. Fluorescence of Ag-Ab complicated is measured, together with scatter houses of cells.

Question five. What Are The Contests Of The Test Tube During Flow Cytometry?

Answer :

Cells from whatever specimen you're testing. Often blood, BM, or LNs in hematopathology. Tissue answers in which cells are disaggregated. Abs with fluorochromes attached to them.

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Question 6. What Happens In The Flow Cytometer?

Answer :

Cells undergo chamber and ideally skip thru 1–via–1. Laser light source shines on cells. If Abs (from reagent you brought) are connected to cells they may supply off their fluorochrome and you will see fluorescence. Detects ahead scatter properties and facet scatter residences on different cells.

Question 7. What Kind Of Data Is Collected From Flow Cytometry?

Answer :

Forward scatter. Side scatter. Fluorescence emitted.

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Question 8. What Does Forward Scatter Tell You?

Answer :

Reflects the scale of the cell. As these one of a kind sized cells skip through Flow Cytometer and cross beyond the laser, laser collects ahead scatter based totally on how plenty mild receives scattered while these cells pass beyond the beam. *Smallest cells = lowest quantity of forward scatter. *Larger cells = maximum ahead scatter.

Question nine. What Does Side Scatter Tell You?

Answer :

Reflects the complexity of the mobile cytoplasm. Neutrophil: many granules → very complex cytoplasm → excessive amount of facet scatter. Reactive lymphocyte: few granules → much less complex cytoplasm → low quantity of side scatter.

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Question 10. During Flow Cytometry, How Do You Assess The Size Of The Cell?

Answer :

Based on Forward Scatter: *Smallest cells = lowest quantity of forward scatter. *Larger cells = most forward scatter.

Question eleven. During Flow Cytometry, How Do You Assess The Complexity Of The Cell Cytoplasm?

Answer :

Based on Side Scatter: *Lowest complexity = lowest facet scatter. *Highest complexity = most aspect scatter.

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Question 12. What Are The Expected Findings For A Lymphocyte?

Answer :

Teal and purple. Low forward scatter (smallest cells in blood and BM). Low side scatter (lowest complexity cytoplasm – no granules).

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Question thirteen. What Are The Expected Findings For Blasts Aka Stem Cells?

Answer :

Red. Higher ahead scatter (very big cells). Low facet scatter (do not have prominent granules).

Question 14. What Are The Expected Findings For Monocytes?

Answer :

Dark blue. Higher forward scatter (larger cells in blood and BM). Intermediate aspect scatter (a few granularity in cytoplasm).

Question 15. What Are The Expected Findings For Granulocytes (neutrophils)?

Answer :

Green. Low to intermediate forward scatter (small to medium length). Very high facet scatter (plenty of granules).

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Question sixteen. What Is On The Axises For A Graph Generated By The Fluorescence?

Answer :

X–axis: wavelength. Y–axis: depth.

Question 17. What Determines The Wavelength?

Answer :

Depends on what fluorochrome you categorised in your reagent. Each fluorescent marker offers of mild at a unique wavelength.

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Question 18. How Do You Deal/regulate For Compensation In Flow Cytometry Of More Than One Wavelength?

Answer :

Conjugate beads to just FITC, and beads to simply PE and then you could subtract for compensation.

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Question 19. When You Use Many Different Ab Reagents Specific For The Cell Types You Want To Identify, What Can Make Understanding Results Difficult?

Answer :

There is a lot of go-reactivity. Cells might have severa amounts of antigens on them. What's expressed at the green cell can also be expressed at the blue cell (so Ab in opposition to inexperienced mobile Ag will also bind to the blue mobile). Cells bypass thru and feature variable quantities of various Abs attached to them.

Question 20. What Would You Expect For Ag Expression In Normal Cells Vs Abnormal Cells?

Answer :

Normal: ought to see very particular Ag expression pattern. Abnormal: inappropriate expression of Ag (eg, blast inappropriately expressing a totally mature marker).

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Question 21. How Do You Interpret This?

Answer :

CD38 (+): something above horizontal line.

CD20 (+): anything to the proper of the vertical line.

Red cells = CD38+ and CD20–.

Blue cells = CD38– and CD20+.

Question 22. What Are The Antibodies Expressed On Granulocytes?

Answer :

CD45 = Leukocyte Common Antigen, LCA.
CD15 = Granulocyte marker.
CD13.
Question 23. What Are The Antibodies Expressed On T Cells?

Answer :

CD45 = Leukocyte Common Antigen, LCA.
CD3 = T–mobile marker.
CD4 = helper T cells.
CD8 = cytotoxic T cells.
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Question 24. What Is Cd3 Expressed On?

Answer :

T cells.

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Question 25. What Is Cd4 Expressed On?

Answer :

Helper T cells.

Question 26. What Is Cd8 Expressed On?

Answer :

Cytotoxic T cells.

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Question 27. What Is Cd13 Expressed On?

Answer :

Granulocytes, monocytes.

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Question 28. What Is Cd14 Expressed On?

Answer :

Monocytes.

Question 29. What Is Cd15 Expressed On?

Answer :

Granulocytes, monocytes.

Question 30. What Is Cd19 Expressed On?

Answer :

B cells.

Question 31. What Is Cd20 Expressed On?

Answer :

B cells.

Question 32. What Is Cd34 Expressed On?

Answer :

Blasts / stem cells.

Question 33. What Is Cd45 Expressed On?

Answer :

All leukocytes.

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Question 34. What Are The Antibodies Expressed On Monocytes?

Answer :

CD13, CD14, CD15 (CD45 - all).

Question 35. What Are The Antibodies Expressed On B Cells?

Answer :

CD19, CD20 (CD45 – all).

Question 36. What Are The Antibodies Expressed On Blasts / Stem Cells?

Answer :

CD34 (CD45 – all).

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Question 37. How Do You Assess B Cell Clonality?

Answer :

Look at their light chain expression patterns.

Question 38. How Do You Assess T Cell Clonality?

Answer :

Express specific antigens, we will appearance carefully at hem to search for clonality.

Question 39. How Do You Assess Plasma Cell Clonality?

Answer :

Expect blend of Abs to be on plasma cells. Expect mix of mild chains (kappa and lambda). When these predicted patterns deviate, can examine/become aware of clonality in these cellular kinds.

Question forty. If You Are Measuring With four "filters", How Many Measurements Will You Have In The End?

Answer :

Six. Forward scatter (length), ninety diploma scatter (inner strx) and the 4 filters (filters have increasingly more better frequency).

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Question 41. What Is "gating"?

Answer :

Selecting to degree information on best one organization of cells.

Question forty two. Can Multiple Antigens Be In The Same Tube?

Answer :

No, however don't ask me what which means: "Each antigen calls for a diff tube".

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