Figure 1. Cytotoxicity test on 96 well plates that was monitored by ELISA reader 3- Discussion 1- Explain another methods by which cytotoxicity is measured ?
Figure 1. Cytotoxicity test on 96 well plates that was monitored by ELISA reader 3- Discussion 1- Explain another methods by which cytotoxicity is measured ?
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Author:Margaret Rodriguez, Paul Price
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![Experiment Title: Cytotoxicity Test
Experiment no. 2
Objectives: Study the cytotoxicity and how it is measured.
1- Background
Cytotoxicity can lead healthy living cells to three potential
cellular fates.
1. Neerosis (accidental cell death): Rapid loss of membrane integrity and cell
lysis
2. Apoptosis (programmed cell death): slower, more orderly, and genetically
controlled
3. Cytostasis (a decrease in cell viability): cells remain alive but fail to
actively grow and divide
Importance of measuring cytotoxicity two major reasons
1. Either you
want specific cells to die
look for an adequa
compound/condition (cancer and immotherapy)
2. Or you want to exclude cyto icity in specific cells (chemicals and drugs)
i have a report on this
subject, please solve
Applications
- Drug discovery proces
- Oncological research
- Safety evaluation of pesti ides, plantextracts, food additives, cosmetics, and
my topic.
industrial chemicals
Classification of cytotoxicity and cell viability assays
These essays are classified according to measurement types of endpoints
(color changes, fluorescence, luminescent, etc.).
1. Dye exclusion: Trypan blue, cosin, Congo red, erythrosine B assays.
2. Colorimetric assays: MTT assay, MTS assay, XTT assay, WST-I assay.
WST-8 assay, LDH assay, SRB assay, NRU assay, and crystal violet assay.
3. Fluorometric assays: alamarBlue assay and CFDA-AM assay.
4. Luminometric assays: ATP assay and real-time viability assay.
2- Experimental Procedure
1- Plate ceils into 96-well tissue culture plates. In general, 5000 - 10,000 ceils
per well according to the cxperimental factors tested. Carry out your
experiment by adding chemicals or biological agents into the appropriate
well. Incubate for 6 to 24 hours.
2- remove 96 well plates from the incubator and add 20 ul MTT stock
solution to each weil
3- place 96 well into an incubator at 37°C, 5% CO2 for 4 hours
4- remove 96 well plates fiom the incubator and aspirate the solution
5- add 100 ul DMSO to each well to dissolve the formazan
6- rotate the plate for 5 min to distribute eveniy
7- Record absorbance at 540nm
Figure 1. Cytotoxicity test on 96 well plates that was monitored by ELISA reader
3- Discussion
1- Explain another methods by which cytotoxicity is measured ?](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F8a7bd013-1c2c-44bf-91c9-42be6d8325bc%2F1fc95de6-392b-4310-a373-13cbe952f6b2%2F9mkg1x_processed.jpeg&w=3840&q=75)
Transcribed Image Text:Experiment Title: Cytotoxicity Test
Experiment no. 2
Objectives: Study the cytotoxicity and how it is measured.
1- Background
Cytotoxicity can lead healthy living cells to three potential
cellular fates.
1. Neerosis (accidental cell death): Rapid loss of membrane integrity and cell
lysis
2. Apoptosis (programmed cell death): slower, more orderly, and genetically
controlled
3. Cytostasis (a decrease in cell viability): cells remain alive but fail to
actively grow and divide
Importance of measuring cytotoxicity two major reasons
1. Either you
want specific cells to die
look for an adequa
compound/condition (cancer and immotherapy)
2. Or you want to exclude cyto icity in specific cells (chemicals and drugs)
i have a report on this
subject, please solve
Applications
- Drug discovery proces
- Oncological research
- Safety evaluation of pesti ides, plantextracts, food additives, cosmetics, and
my topic.
industrial chemicals
Classification of cytotoxicity and cell viability assays
These essays are classified according to measurement types of endpoints
(color changes, fluorescence, luminescent, etc.).
1. Dye exclusion: Trypan blue, cosin, Congo red, erythrosine B assays.
2. Colorimetric assays: MTT assay, MTS assay, XTT assay, WST-I assay.
WST-8 assay, LDH assay, SRB assay, NRU assay, and crystal violet assay.
3. Fluorometric assays: alamarBlue assay and CFDA-AM assay.
4. Luminometric assays: ATP assay and real-time viability assay.
2- Experimental Procedure
1- Plate ceils into 96-well tissue culture plates. In general, 5000 - 10,000 ceils
per well according to the cxperimental factors tested. Carry out your
experiment by adding chemicals or biological agents into the appropriate
well. Incubate for 6 to 24 hours.
2- remove 96 well plates from the incubator and add 20 ul MTT stock
solution to each weil
3- place 96 well into an incubator at 37°C, 5% CO2 for 4 hours
4- remove 96 well plates fiom the incubator and aspirate the solution
5- add 100 ul DMSO to each well to dissolve the formazan
6- rotate the plate for 5 min to distribute eveniy
7- Record absorbance at 540nm
Figure 1. Cytotoxicity test on 96 well plates that was monitored by ELISA reader
3- Discussion
1- Explain another methods by which cytotoxicity is measured ?
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