Hello sir, I khow it lakes preciouS Lab. Sheet-Level time, Please, I have a report on this Semester 1 subject. Only 5 papers within a Experiment Title: Cytotoxicity Test discussion solution. I want to solve the text of the keyboard. Thank you, Experiment no. 2 sir 1 12:34 PM Objectives: Study the cytotoxicity and how it is measured. 1-Background Cytotoxicity can lead healthy living cells to three potential cellular fates 1. Necrosis (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 and look for an adequate compound/condition (cancer and immunotherapy) 2. Or you wwant to exclude cytotoxicity in specifc cells (chemicals and drugs) Applications - Drug discovery process - Oncological research - Safety evaluation of pesticides, plant extracts, food additives, cosmetics, and 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. Colorinmetric 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 cells into 96-well tissue culture plates. In general, 5000 -10,000 cells per well according to the experimental 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 l MTT stock solution to each well 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 DMSC to each well to diasolve the formazan 6- rotate the plate for 5 min to distribute evenly 7- Record absorbance at 540nm
Hello sir, I khow it lakes preciouS Lab. Sheet-Level time, Please, I have a report on this Semester 1 subject. Only 5 papers within a Experiment Title: Cytotoxicity Test discussion solution. I want to solve the text of the keyboard. Thank you, Experiment no. 2 sir 1 12:34 PM Objectives: Study the cytotoxicity and how it is measured. 1-Background Cytotoxicity can lead healthy living cells to three potential cellular fates 1. Necrosis (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 and look for an adequate compound/condition (cancer and immunotherapy) 2. Or you wwant to exclude cytotoxicity in specifc cells (chemicals and drugs) Applications - Drug discovery process - Oncological research - Safety evaluation of pesticides, plant extracts, food additives, cosmetics, and 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. Colorinmetric 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 cells into 96-well tissue culture plates. In general, 5000 -10,000 cells per well according to the experimental 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 l MTT stock solution to each well 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 DMSC to each well to diasolve the formazan 6- rotate the plate for 5 min to distribute evenly 7- Record absorbance at 540nm
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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Transcribed Image Text:Biomedical Engincering Dept.
Biocompatibility Hello sir, I know it takes precious
Lab. Sheet-Level time. Please, I have a report on this
Semester 1
subject. Only 5 papers within a
Experiment Title: Cytotoxicity Test discussion solution. I want to solve
the text of the keyboard. Thank you,
Experiment no. 2
sir
1 12:34 PMA
Objectives: Study the eytotoxicity and how it is measured.
1- Background
Cytotoxicity can lead healthy living cells to three potential
cellular fates.
1. Necrosis (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 and look for an adequate
compound/condition (cancer and immunotherapy)
2. Or you want to exclude cytoloxicity in specific cells (chemicals and drugs)
Applications
- Drug discovery process
- Oncological research
- Safety evaluation of pesticides, plant extracts, food additives, cosmetics, and
industrial chemicals
Classification of cytotoxicity ad 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. Colorinmetric 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 cells into 96-well tissue cuiture plates. In general, 5000 -10,000 ceils
per well according to the experimental factors tested. Carry out your
experiment by adding chemicalIs or biological agents into the appropriate
well, Incubate for 6 to 24 hours,
2- remove 96 well plates fiom the incubator and add 20 pul MTT stock
solution to each wel
3- place 96 well into an incubator at37 °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 dissolvec the formazan
6- rotate the plate for 5 min to distribute evenly
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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