really shitty biotech review for flow cytometry part 2 (rhyme for the mixtape)

Question 1

Monocytes originate in the _____ ______ from a rapidly growing pool of myeloid progenitor cells called __________.

Answer 1

1 - bone marrow

2 - promyelocytes

Question 2

Monocytes are a type of ________ (WBC) that constitute the basic cellular components of the ________ blood in which they circulate in and make up between ___ and ___ % of the total WBCs in the body.

Answer 2

1 - leukocyte
2 - peripheral
3 - 5;10

Question 3

After a short _______ ______ of 1-3 days, they then circulate briefly in the blood and migrate into the _______ where they assume their ultimate role as ________.

Answer 3

1 - developmental period
2 - tissues
3 - macrophages

Question 4

_____ - ______ macrophages on the other hand, reside in the the tissue and are dedicated to fulfilling tissue specific functions including?

Answer 4

1 - Tissue resident
Functions include: clearance of cellular debris, play a central role in immune surveillance and the inflammatory response.

Question 5

Macrophages respond in two primary ways. These are?

Answer 5

chemoattraction and adhesion.

Question 6

What is chemoattraction?

Answer 6

When macrophages in the affected area release cytokines. This in turn causes the endothelial cells near the area, to express cellular adhesion molecules resulting in the localization of circulating leukocytes to the site of injury or infection.
To sum up:
macrophages release cytokines –> causes endothelial cells to express cellular adhesion molecules -> result is localization of circulating leukocytes to site of injury/infection

Question 7

What are examples of cytokines?

Answer 7

IL-1 and TNF-alpha

Question 8

What are examples of cellular adhesion molecules?

Answer 8

P- and E-selectin

Question 9

What are endothelial cells?

Answer 9

cells that line blood vessels

Question 10

What is the mechanism of adhesion?

Answer 10

Macrophages release chemokines (oh yeah thx scientists, put chemokines with adhesion rather than chemoattraction…) –> causes surface integrin molecules on the endothelial cells to kick into overdrive (definitely going in the lab report) going into a high-affinity state. –> causes immobilization of leukocytes to that area (Since integrins become tightly bound to complementary receptors on the endothelial cells) –> they then enter the tissues as an influx in order to differentiate and replenish tissue-specific macrophages

Question 11

The differentiation process is due to what?

Answer 11

Result of signals from locally produced factors such as M-CSF, which prepares the cells to actively participate in teh inflammatory and innate immune response.

Question 12

Upon differentiation, the cells lose their ability to _______ leading to enhanced ___________ activity enabling them to _______ and _______ (or _________) foreign material, dead cells, etc.

Answer 12

1 - replicate
2 - antibacterial
3 - engulf and digest or phagocytose

Question 13

M-CSF stands for?

Answer 13

Macrophage - colony stimulating factor

Question 14

What are examples of integrin molecules?

Answer 14

VCAM-1 and alpha4beta1

Question 15

The human monocytic leukemia cell line, ____, can be induced to differentiate into ________ by treatment with ___ or _____.

Answer 15

1 - THP-1
2 - macrophages
3 - PAM or Vit-D3

Question 16

Primary tissue macrophages are not readily expandable in _____ systems.

Answer 16

in-vivo

Question 17

What does not expandable mean?

Answer 17

cannot grow them easily in large quantities

Question 18

The _________ state and _________ of this cell line however can vary based on the type of _________ and _______ used to induce differentiation and can therefore only moderately predict the behaviour of typical ________ ________ in living systems.

Answer 18

1 - differentiation state
2 - phenotype
3 - treatment
4 - duration
5 - primary macrophages

Question 19

Differentiation of monocytic cell lines into highly differentiated macrophages is a primary focus in order to closely resemble what?

Answer 19

MDM - monocyte derived macrophages

Question 20

Further culture of ____ cells following treatment with ____ in regular _____, drives cells more towards the differentiation states of typical ____.

Answer 20

1 - THM-1
2 - PAM
3 - media
4 - MDMs

Question 21

The expansion of the ________ is central to the function of differentiated _________ as this enhances their ________ capacity allowing for more cellular debris and foreign microbes to be cleared from the tissue.

Answer 21

1 - cytoplasm
2 - macrophages
3 - phagocytic

Question 22

Further analysis by flow cytometry generally detects an observable increase in _____ _____ or _____/________.

Answer 22

1 - side scatter

2 - granularity/density

Question 23

____ treatment also leads to a more mature ________ resulting in a slower rate of ________, higher levels of _______, higher rate of _________ in addition to an increased cell ______ expression of ____ and _____.

Answer 23

1 - PMA
2 - phenotypes
3 - proliferation
4 - phagocytosis
5 - adherence
6 - surface
7 - Cd14 and CD11b

Question 24

Classically, an increase in _____ expression has been widely used as a factor to identify _______ of monocytes to macrophages however, more recently, scientists have discovered that the _____ culture conditions (______ and _______ length) of ____ monocytes, prior to any stimuli (such as ____ treatment), is a critical factor that can influence the _______ response and ____ of macrophages.

Answer 24

1 - CD14
2 - differentiation
3 - basal
4 - confluence and culture length
5 - THP-1
6 - PMA
7 - differentiated
8 - state

Question 25

What is another well used approach for the classification and identification of macrophages?

Answer 25

Autofluorescence.

Question 26

The full biological mechanism behind the resultant autofluorescence is not yet fully understood however, it has been suggested that it could be derived as a result of what?

Answer 26

increased complexity in the cells, flavins and/or cellular lipids

Question 27

In this lab you will use ____ ______ as a means to examine the characteristic differences between ________ and _________ in ____ treated ____ cells.

Answer 27

1- flow cytometry
2 - monocytes and macrophages
3 - PMA treated THP-1

Question 28

We will be analyzing the _____ light ____ and ____ _____ plots to identify features and differences in ____ and _______ of monocytes compared to macrophages.

Answer 28

1 - forward and side scatter

2 - size and granularity

Question 29

How can autofluorescence be measured?

Answer 29

by the FL1 channel

Question 30

The lysosome concentration should increase with _______ due to an increase in ______ volume, measured by the ___ channel.

Answer 30

1 - differentiation (monocytes –> macrophages)
2 - cytoplasm
3 - FL3

Question 31

The different expression of cell-surface antibodies is measured by what?

Answer 31

1 - CD14-FITC via the FL1 channel

2 - CD4-Texas red via the FL3 channel

Question 32

The phagocytic capacity of THP-1 cells should also _____ with differentiation due to an overall increase in ____ _____ and ______ and will be monitored through the ___ channel.

Answer 32

1 - increase
2 - size and volume
3 - FL3