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13 Cards in this Set
- Front
- Back
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Flow cytometry is a measure of two things, what are they?
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Light scattered by cells and fluorescence from fluorochromes attached to cells.
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How do cells flow through the central core?
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In single file
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Does the sample fluid mix with the sheath fluid?
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kNOw
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Fluorochromes bound to cells are excited by the laser and emit light at higher or lower wavelengths?
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Higher
Laser is usually at wavelenght of 488nM |
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How do you deal/adjust for compensation in flow cytometry of more than one wavelength?
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Conjugate beads to just FITC, and beads to just PE and then you can subtract for compensation.
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In flow cytometry are you measuring in all blood cells?
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No, you lyse the RBC's prior to the procedure.
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If you are measuring with 4 "filters", how many measurements will you have in the end?
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Six. Forward scatter (size), 90 degree scatter (internal strx) and the four filters.
(filters have increasingly higher frequency.) |
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How is the graph set up for flow cytometry?
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Horizontal axis (x) is the fluorescence intensity, and the vertical (y) axis and the cell number.
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What is "Gating"
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Selecting to measure info on only one group of cells.
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Can multiple antigens be in the same tube?
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No, but don't ask me what that means:)
"Each antigen requires a diff tube" |
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Light is collected at 90degrees and:
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In front (forward scatter)
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More forward scatter, means:
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A larger cell
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1. Small, (so little forward scatter), no granules or segmentation (so not much 90degree scatter).
2. Larger, more internal strx 3. Larger, segmented, more granules (so more 90 degree scatter) |
1. Lymphocytes
2. Monocytes 3. Granulocytes |
