Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Immunology final > Practice #4 > Flashcards

Practice #4 Flashcards

1
Q

Flow cytometry/ FACS

A

Fluorescence activated cell sorting

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Advantages of flow cytometry as oppose to microscope

A
  1. Objective 2. Large num of cells can be examined in 1 measurment 3. Automatic 4. Fast
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

The basic structure of flow cytometer

A
  1. Fluidics: cell flow and hydrodynamic focusing to the point of detection 2. Optics: Laser beam is concentrated and passes through one cell at the time on the other side the emitted light is detected by FSC (front scatter detector) and a series of SSC (side scatter detectors) 3. Electronics: transforming light signals to electric ones for FSC its photodiode and for SSC its PMT 4. Analysator unit: collection, storage and processing
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Hydrodynamic focusing is done by

A
  1. The shape of the flow cell (funnel shape) 2. Sheath fluid (isitonic buffer) : flowing in laminar flow to the opposite direction of the sample stream
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What are the functions of FSC and SSC?

A

FSC - Indicator of the cell size with direct proportionality SSC - Indicator for the complexity of the cell (granular or not)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What are the possible fluorescent markers used to tag the cell

A

Direct, indirect, biotin-avidin, intracellular molecules

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Possible data imaging methods

A
  1. Histogram - one single parameter depending on cell count 2. Dot plot - 2 or more parameters
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

Applications of flow cytometry

A
  1. cell sorting 2. total cell count 3. detecting different cell populations in the sapmle 4. detection of the functional state of cells in the sample
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

Preparation of the sample for flow cytometry

A
  1. Fresh suspension (not coagulated blood) 2. Erythrolysis 3. Staining if necessary
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Dot plot complexity vs. size

A
  1. Eosinophils
  2. Neutrophils
  3. Monocytes
  4. Lymphocytes and Basophils
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

Dot plot diagnosis

A

Left: Normal bone marrow smear

Right: Bone marrow leukemia

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

What happened between A and B?

A

In A there is more fluorescence proteins attached to each cell

Left shift

(intensity shift)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

What happened between A and B?

A

In B the number of cells expressing CD3 is increased

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

What are the different quality check possibilities in flow cytometry?

A
  1. Technical:

setting the machine

Fluorescence compensation for molecules with lower intensity fluorescence

  1. Biological:

Unstained preparation

or

Unspecific binding to control isotope

Detection of autofluorescence

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Markers for NKC

A

CD3-

CD56+

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Markers for B cells

A

CD19+

17
Q

Markers for B1 cells

A

CD5+

18
Q

Markers for plasma cells

A

ic light chain +

CD19-

19
Q

Markers for Th cells

A

CD3+

CD4+

20
Q

Markers for T cells

A

CD3+

21
Q

Markers for NKT cells

A

CD3+

CD56+

22
Q

Markers for Tc cells

A

CD3+

CD8+

23
Q

Markers for gamma/delta T cells

A

CD3+

gamma/delta TCR +

CD4-

CD8-

24
Q

Markers for Treg cells

A

CD4+

CD25++

Foxp3+

25
Q

Markers for Th1 cells

A

IFNgamma+

IL12+

TNFalpha+

TNFbeta+

26
Q

Markers for Th2 cells

A

IL4+

IL5+

IL10+

IL13+

27
Q

Markers for Th17 cells

A

IL17+

TGFbeta+

28
Q

Markers for lymphoid progenitor cells

A

CD34+

cKit+

HLA-DR+

TdT+

29
Q

Soluble cytokine detection by flow cytometry

A

A microbead with specific Ab against the cytokine is mixed with the sample

then a secondary Ab (with fluorescence dye) against the cytokine is added

30
Q

Cell cycle analysis by flow cytometry

A
  1. membrane lysis
  2. DNA staininng by propium iodide

* elevation of the S is an indicatorfor malignancy

31
Q

Phagocyte function analysis by flow cytometry

A

Phagosytosis of fluorescnece labled latex beads will release the dye and the intensity of fluorescence will be higher

(the x axis on the right the y axis on the left)

32
Q

Cell sorting by flow cytometry

A

Two possibilities:

  1. Mechanic separation by laser interrogation
  2. Electrostatic separation
33
Q

Acute myeloid leukemia markers

A

CD14+

HLD-DR+

CD13+

CD33+

icMPO+

34
Q

Acute lymphoid leukemia markers

A

HLA-DR+

icTdT+

Non T all

CD19+ CD20+

(CALLA)

CD10+

T all

CD2+ CD3+ CD5+ CD7+

35
Q

CD45

A

Present on all WBC except plasma cells

36
Q

CD38

A

Typical activation marker of Th, Tc, B, NKC

Prognostic marker for leukemia

37
Q

CD138

A

Highly expressed on all tumor cells

38
Q

FCM in pharmacology

A

Pgp detection by immunophenotyping

Measure the time it takesto the cell to pump out the fluorescent labled drug

39
Q
A

Immunology final (24 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions