CELL BIOLOGY LAB FINAL
Where do you dispose of broken glass that has not come in direct contact with a specimen? –
Broken glass waste bin
If you wanted to see the cells present in a culture plate without disturbing the cells, in the plate,
which microscope would you use? – Inverted microscope
As magnification increases, field of view – decreases.
As magnification increases, depth of field – decreases.
While looking at a cell sample, you are trying to locate the lysosome that is 396 nm in diameter.
Which objective would you use to resolve the organelle? - 100x - resolution 259 nm
Which of the following is NOT a connective tissue? – muscle.
Which type of tissue is specialized for secretion and absorption? – epithelium
In today's experiment, you would look at tissue slides to determine the identity of the tissue
samples. Which microscope would you use to accomplish this? - compound light microscope
Name a type of nervous tissue – neurons
What is a morphological difference between smooth muscle and skeletal muscle? - smooth
muscle does not have any striations, while skeletal muscle does
Where can cardiac muscle be found? – heart
Epithelial tissue function - responsible for absorption, secretion, and protection
Muscular tissue function – specialized for contraction
Nervous tissue function - transport of electrochemical signals in the body
Connective tissue function - abundance of extracellular matrix (ECM), provides organ support
Adipose is a type of ___ which is ___. - connective tissue; used for energy metabolism and
insulation
Hyaline cartilage is a type of epithelial tissue – false
What is the point of using an incubator? - -to maintain optimal temperature for cells, -to control
carbon dioxide content of the atmosphere inside, -to maintain optimal humidity for cells
Why are cells grown in specialized culture media? - specialized culture medias are used to
guarantee adequate reproducibility that natural medias lack
It's important to set the incubator at a very high temperature, so the cells don't die – False
A biological safety cabinet is used to - protect yourself from harmful agents inside the cabinet,
protect the organisms you're working with from contamination, protect the environment from
harmful contaminants.
What is a hemocytometer use for? – to count cells
How would you know it is time to change the growth media in a cell culture dish? - waste
produced by the cells disrupts the pH of the media
What is the purpose of adding trypsin to your cell plate? - detach cells from the bottom of the
plate
What dye would you use if you wanted to stain dead cells? - trypan blue
After using a hemocytometer to count fibroblast cells, you find a concentration of 500,000
cells/mL. You now want to split cells and make a new culture that contains 250,000 cells. How
many microliters from your original culture would you have to add? – 250,000/500,000= 0.5
cells/ml. Then convert cells/ml into cells/ul. 0.5*1000 = 0.5 ul
Calculate the percent viability of a sample that has 45 white cells and 75 blue cells (120 cells
total) – 37.5
, Mounting is the process of adding a chemical fixative that cross links proteins into rigid lattice
structures – False
Name two of three different types of cell staining methods (detection
methods) - antibody staining, fluorescent staining
What does the process of permeabilization do to the cells? - dissolve cell membranes
How would you differentiate a primary from a secondary antibody? - Primary antibodies that
bind directly to the target will have a fluorophore attached.
Secondary antibodies with an attached fluorophore bind to the primary antibody which binds to
the target. Indirect immunofluorescence with the use of secondary antibodies are brighter.
In today's lab, we utilized Triton X instead of Digitonin to fully permeabilize the inner membrane
because Digitonin is only able to penetrate the outer membrane. Had we used Digitonin instead,
what dye would have not worked at all? – DAPI
Name one difference in the staining pattern observed between the two cells
(Dictyosteliumdiscoideum vs. L-fibroblasts) - Dicty are more grouped together and smaller, while
the Fibroblasts were larger, more spread out, and showed more of its structure
DAPI function - binds to the nucleus and dyes fluorescent blue
Paraformaldehyde function - used to "fix" or preserve cell morphology
Fluorescein-phalloidin conjugate function - binds to actin filaments, carries fluorescent green
dye
Triton X function - permeabilizes both inner and outer membranes
The nutritional status of the Dictyostelium cells has no impact on whether they would prefer
folic acid or cAMP as a chemoattractant – False
Waves of CAMP cause the Dictyosteliumcells to respond on a ___ level – colonial
Based on the relative quick speed of cell movement observed this week (under 3 hours),we
know that Dictyostelium utilize ___ cell migration – amoeboid
What question were we trying to answer (Chemotaxis lab) today? - Which chemoattractant will
Dictyostelium cells preferably chemotaxis to, cAMP or folic acid? Will they chemotaxis to them
at all?
What cells did we work with today? (Chemotaxis) – Dictyostelium
The two main types of cell migration are - mesenchymal and amoeboid
Chemotaxis specifically refers to the directed migration of a cell or organism specifically toward
an external chemical stimulus – False
Name 2 of the 3 chemoattractants we tested - cAMP and folate
What was the purpose of the Endocytosis experiment? - We want to determine what type of
endocytosis THP1 macrophage human cells utilize to engulf E. coli bacteria with two treatments:
Lat A and DMSO
What was Triton-X used for in the endocytosis lab? - cell membrane permeabilizer
Phagocytosis results in the cytoskeletal rearrangement of tubulin in order to envelop particles –
False
Name one type of micropinocytic mechanism - clathrin-mediated endocytosis
The "large scale endocytic processes" (engulf particles larger than .2 um) are - phagocytosis and
macropinocytosis
What three things were fluorescently stained in today's experiment? - cell membranes, cell
nucleus, and bacterial cells
, Why did you incubate your cells at 4C after adding bacteria to the culture dish? (Endocytosis lab)
- to stop the metabolism of the cells and let the bacteria settle on them
What was the control used in the Endocytosis lab? Why was it chosen as our control? - DMSO. It
was chosen because Lat A is in (and is soluble) 0.1% DMSO. Having DMSO as our control can
show us the effects of Lat A.
When selecting macrophages to count bacteria, it was very important to select ___ to ensure
accurate results – random cells
What is Wheat Germ Agglutinin and what did we use it for today? (Endocytosis lab) - it is a red
fluorescent stain; we used it to stain the cell membrane
What is the question we were trying to answer in the Adipogenesis lab? - What is the effect of
insulin concentration on adipogenesis in white adipose tissue?
What are the two types of adipose tissue? – white, brown
What 3 compounds were important to add to the culture media to induce adipogenesis? -
insulin, DEX, & IBMX
What did you use to stain adipocytes? – oil red-o
What type of cells were we working with in the Adipogenesis lab? – fibroblast
To stimulate adipogenesis on the first day, cells were treated with – differentiation media
If the absorbance value for your sample goes above 0.9 you must - dilute your sample because it
is too concentrated
Using the vacuum trap on a lower power was very important in the Adipogenesis lab because -
adipose tissue is more viscous than other tissues
(Adipogenesis lab) When analyzing our data, why did we make a second graph in which the
value for 0mg/ml of insulin was graphed at 0.0? - in order to see the direct effect insulin
concentration has without the effects of iBMX and DEX
How did the elution process to extract Oil Red O (ORO) from the adipocytes work? - Isopropanol
was added to the dish to extract ORO, since isopropanol is more nonpolar than the fat inside the
adipocytes
During the last lab, students used HRP enzyme to catalyze the reaction between o-
phenylenediamine and H2O2. After 8 minutes of incubation in the dark, the students used what
to stop the reaction? Why was this reagent successful in stopping the enzymatic reaction? -
sulfuric acid, denatures the enzyme and stops the reaction
Chlorpromazine (endocytic pathway?) - clathrin micropinocytosis
Nystatin (endocytic pathway?) - lipid raft-caveolae micropinocytosis
Amiloride (endocytic pathway?) – macropinocytosis
T/F The accumulation of cholesterol inside of the cells transforms macrophages into the so-
called phagocytic cells – False
Atherosclerosis is a disease in which the inside of the arteries narrows after a buildup of ___
(organic compound) - LDL-c -holesterol
What type of cell line that was isolated from a sick patient with cholesterol plaque accumulation
will we be testing today? - fibroblast cells
What is the purpose of today's lab (endocytosis)? - to determine which endocytic mechanism is
responsible for the excessive uptake of LDL-cholesterol
What cell signaling pathway is inhibited by Trametinib? – RTK-Ras
What cell signaling pathway is inhibited by SQ536? – gPCR
Where do you dispose of broken glass that has not come in direct contact with a specimen? –
Broken glass waste bin
If you wanted to see the cells present in a culture plate without disturbing the cells, in the plate,
which microscope would you use? – Inverted microscope
As magnification increases, field of view – decreases.
As magnification increases, depth of field – decreases.
While looking at a cell sample, you are trying to locate the lysosome that is 396 nm in diameter.
Which objective would you use to resolve the organelle? - 100x - resolution 259 nm
Which of the following is NOT a connective tissue? – muscle.
Which type of tissue is specialized for secretion and absorption? – epithelium
In today's experiment, you would look at tissue slides to determine the identity of the tissue
samples. Which microscope would you use to accomplish this? - compound light microscope
Name a type of nervous tissue – neurons
What is a morphological difference between smooth muscle and skeletal muscle? - smooth
muscle does not have any striations, while skeletal muscle does
Where can cardiac muscle be found? – heart
Epithelial tissue function - responsible for absorption, secretion, and protection
Muscular tissue function – specialized for contraction
Nervous tissue function - transport of electrochemical signals in the body
Connective tissue function - abundance of extracellular matrix (ECM), provides organ support
Adipose is a type of ___ which is ___. - connective tissue; used for energy metabolism and
insulation
Hyaline cartilage is a type of epithelial tissue – false
What is the point of using an incubator? - -to maintain optimal temperature for cells, -to control
carbon dioxide content of the atmosphere inside, -to maintain optimal humidity for cells
Why are cells grown in specialized culture media? - specialized culture medias are used to
guarantee adequate reproducibility that natural medias lack
It's important to set the incubator at a very high temperature, so the cells don't die – False
A biological safety cabinet is used to - protect yourself from harmful agents inside the cabinet,
protect the organisms you're working with from contamination, protect the environment from
harmful contaminants.
What is a hemocytometer use for? – to count cells
How would you know it is time to change the growth media in a cell culture dish? - waste
produced by the cells disrupts the pH of the media
What is the purpose of adding trypsin to your cell plate? - detach cells from the bottom of the
plate
What dye would you use if you wanted to stain dead cells? - trypan blue
After using a hemocytometer to count fibroblast cells, you find a concentration of 500,000
cells/mL. You now want to split cells and make a new culture that contains 250,000 cells. How
many microliters from your original culture would you have to add? – 250,000/500,000= 0.5
cells/ml. Then convert cells/ml into cells/ul. 0.5*1000 = 0.5 ul
Calculate the percent viability of a sample that has 45 white cells and 75 blue cells (120 cells
total) – 37.5
, Mounting is the process of adding a chemical fixative that cross links proteins into rigid lattice
structures – False
Name two of three different types of cell staining methods (detection
methods) - antibody staining, fluorescent staining
What does the process of permeabilization do to the cells? - dissolve cell membranes
How would you differentiate a primary from a secondary antibody? - Primary antibodies that
bind directly to the target will have a fluorophore attached.
Secondary antibodies with an attached fluorophore bind to the primary antibody which binds to
the target. Indirect immunofluorescence with the use of secondary antibodies are brighter.
In today's lab, we utilized Triton X instead of Digitonin to fully permeabilize the inner membrane
because Digitonin is only able to penetrate the outer membrane. Had we used Digitonin instead,
what dye would have not worked at all? – DAPI
Name one difference in the staining pattern observed between the two cells
(Dictyosteliumdiscoideum vs. L-fibroblasts) - Dicty are more grouped together and smaller, while
the Fibroblasts were larger, more spread out, and showed more of its structure
DAPI function - binds to the nucleus and dyes fluorescent blue
Paraformaldehyde function - used to "fix" or preserve cell morphology
Fluorescein-phalloidin conjugate function - binds to actin filaments, carries fluorescent green
dye
Triton X function - permeabilizes both inner and outer membranes
The nutritional status of the Dictyostelium cells has no impact on whether they would prefer
folic acid or cAMP as a chemoattractant – False
Waves of CAMP cause the Dictyosteliumcells to respond on a ___ level – colonial
Based on the relative quick speed of cell movement observed this week (under 3 hours),we
know that Dictyostelium utilize ___ cell migration – amoeboid
What question were we trying to answer (Chemotaxis lab) today? - Which chemoattractant will
Dictyostelium cells preferably chemotaxis to, cAMP or folic acid? Will they chemotaxis to them
at all?
What cells did we work with today? (Chemotaxis) – Dictyostelium
The two main types of cell migration are - mesenchymal and amoeboid
Chemotaxis specifically refers to the directed migration of a cell or organism specifically toward
an external chemical stimulus – False
Name 2 of the 3 chemoattractants we tested - cAMP and folate
What was the purpose of the Endocytosis experiment? - We want to determine what type of
endocytosis THP1 macrophage human cells utilize to engulf E. coli bacteria with two treatments:
Lat A and DMSO
What was Triton-X used for in the endocytosis lab? - cell membrane permeabilizer
Phagocytosis results in the cytoskeletal rearrangement of tubulin in order to envelop particles –
False
Name one type of micropinocytic mechanism - clathrin-mediated endocytosis
The "large scale endocytic processes" (engulf particles larger than .2 um) are - phagocytosis and
macropinocytosis
What three things were fluorescently stained in today's experiment? - cell membranes, cell
nucleus, and bacterial cells
, Why did you incubate your cells at 4C after adding bacteria to the culture dish? (Endocytosis lab)
- to stop the metabolism of the cells and let the bacteria settle on them
What was the control used in the Endocytosis lab? Why was it chosen as our control? - DMSO. It
was chosen because Lat A is in (and is soluble) 0.1% DMSO. Having DMSO as our control can
show us the effects of Lat A.
When selecting macrophages to count bacteria, it was very important to select ___ to ensure
accurate results – random cells
What is Wheat Germ Agglutinin and what did we use it for today? (Endocytosis lab) - it is a red
fluorescent stain; we used it to stain the cell membrane
What is the question we were trying to answer in the Adipogenesis lab? - What is the effect of
insulin concentration on adipogenesis in white adipose tissue?
What are the two types of adipose tissue? – white, brown
What 3 compounds were important to add to the culture media to induce adipogenesis? -
insulin, DEX, & IBMX
What did you use to stain adipocytes? – oil red-o
What type of cells were we working with in the Adipogenesis lab? – fibroblast
To stimulate adipogenesis on the first day, cells were treated with – differentiation media
If the absorbance value for your sample goes above 0.9 you must - dilute your sample because it
is too concentrated
Using the vacuum trap on a lower power was very important in the Adipogenesis lab because -
adipose tissue is more viscous than other tissues
(Adipogenesis lab) When analyzing our data, why did we make a second graph in which the
value for 0mg/ml of insulin was graphed at 0.0? - in order to see the direct effect insulin
concentration has without the effects of iBMX and DEX
How did the elution process to extract Oil Red O (ORO) from the adipocytes work? - Isopropanol
was added to the dish to extract ORO, since isopropanol is more nonpolar than the fat inside the
adipocytes
During the last lab, students used HRP enzyme to catalyze the reaction between o-
phenylenediamine and H2O2. After 8 minutes of incubation in the dark, the students used what
to stop the reaction? Why was this reagent successful in stopping the enzymatic reaction? -
sulfuric acid, denatures the enzyme and stops the reaction
Chlorpromazine (endocytic pathway?) - clathrin micropinocytosis
Nystatin (endocytic pathway?) - lipid raft-caveolae micropinocytosis
Amiloride (endocytic pathway?) – macropinocytosis
T/F The accumulation of cholesterol inside of the cells transforms macrophages into the so-
called phagocytic cells – False
Atherosclerosis is a disease in which the inside of the arteries narrows after a buildup of ___
(organic compound) - LDL-c -holesterol
What type of cell line that was isolated from a sick patient with cholesterol plaque accumulation
will we be testing today? - fibroblast cells
What is the purpose of today's lab (endocytosis)? - to determine which endocytic mechanism is
responsible for the excessive uptake of LDL-cholesterol
What cell signaling pathway is inhibited by Trametinib? – RTK-Ras
What cell signaling pathway is inhibited by SQ536? – gPCR
