Experiment 3: Cell Viability Testing and Counting with Trypan Blue Exclusion Method
Anna Coretta R. Santos
I. Introduction
Cell viability assay plays a crucial role is in the cells of the cell culture. This assay can be used to identify the relationship of cell count from the cell’s behavior (Stoddart 2011). This test help researchers to determine the number of viable and non-viable cells in a cell culture. This assay helps in estimating the cell count of a cell culture and to determine if there is sufficient amount of viable cells in the cell culture that a person will use for future research (Riss et al. 2013).
A vital tool used in cell viability testing is a hemocytometer. This counting chamber is used for counting cells and helps researchers
…show more content…
Conclusion and Recommendations
Ultimately, testing cell viability and knowing the count of viable and non-viable cells in a cell line is an important key in any research using cells. Having enough number of viable cells in a suspension would give accurate results in an experiment. This also shows how trypan blue is an important dye in cell viability and in the study of cells. This dye makes researcher’s life convenient in identifying and differentiating viable versus non-viable cells or live versus dead cells.
It is really needed that researchers should ensure new and fresh cell cultures to ensure more viable cells in a culture. It also better to ensure that the materials like hemocytometer and pipettes are sterilized and clean so that there would be lesser or no artifacts would be seen under the microscope. The researcher was to use trypan blue exclusion method to test for cell viability, observe the non-viable and viable cells, and was able to have a cell count using the
…show more content…
[Internet]. [Updated: 2012 Aug 10]. Houston: Rice University. [cited 2017 Feb 4]. Available from http://www.ruf.rice.edu/~bioslabs/methods/microscopy/cellcounting.html
Riss, T., Moravec, R., Niles, A., Duellman, S., Benink, H., Worzella, T., Minor, L. 2013. [Internet]. Cell Viability Assays. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences. [cited 2017 Feb 4]. Available from https://www.ncbi.nlm.nih.gov/books/NBK144065/
Stoddart, M. 2011. Cell viability assay: Introduction. Methods in Molecular Biology 740: 1-6.
ThermoFisher Scientific. [Internet]. Masasachusetts: ThermoFisher Scientific. 2016 Dec- [cited 2017 Feb 2]. Available from https://www.thermofisher.com/ph/en/home/life-science/cell-analysis/flow-cytometry/cell-health-and-viability-assays-for-flow-cytometry/cell-viability-assays-for-flow-cytometry.html
Tran, S., Puhar, A., Camus, M., Ramarao, N. 2011. Trypan Blue Dye Enters The Viable Cells Incubated with the Pore-Forming Toxin Hlyll of Bacillus cereus. PLoS One 6(9).
Yip, D., Auersperg, N. 1972. The dye exclusion test for cell viability: Persistence of Differential staining following fixation. In Vitro 7(5): 323-329.
V. Answer to Guide
Last test, inoculation of phenylalanine agar is used to determine if phenylalanine deaminase oxidizes phenylalanine into phenylpyruvic acid and ammonia. Sixth test, is a Multiple Test Media used to determine the physiological characteristics of unknown #398. First test, Inoculation of Kligler 's Iron agar was used to determine the production of hydrogen sulfide from cysteine and fermentation of glucose and lactose. Last test, inoculation of litmus milk is used to determine the fermentation of lactose, casein, lactalbumin, and
Problem 3 solutions using different approaches in the literature Approach Reference Machine cells Number of intercellular
These cells have been used in countless biomedical research efforts, are actively used in laboratories
WARD’S SIMULATED BLOOD Objective: The objective of this lab is to use Ward’s simulated blood to know the blood types and Rh factor of four people. Hypothesis: Based on the exercise, if Mr. Smith is a blood type A+, then he will coagulate with anti A and anti Rh. If Mr jones is a blood type B-, then he will coagulate with Anti- B serum. If Mr Green is a AB+, then he will coagulate with Anti- A, B and Rh.
Starch solution is then placed into the test tube at a quantity of 5 mL. 5 drops of Lugol’s Iodine solution is added to the test tube. If the color changes, then it is known that starches are present in the solution. Proteins are next tested. In order to do this, 5 mL of gelatin solution is added to the test tube. 10 drops of Biuret’s reagent are added to test for protein.
These cells have been the oldest and most commonly used human cell line, and it is also remarkably durable and prolific
LABORATORY REPORT EXERCISE #5 INTRODUCTION TO THE COMPOUND LIGHT MICROSCOPE, PLANT AND ANIMAL CELLS Name_______________________________Section_____Teacher______________Date________ PRE-LAB QUESTIONS - answer the following questions using your textbook and valid internet sources. Be sure to cite your sources at the end of the prelab. You can type your answers to all questions except #1 and #9 directly into this document and then submit via Canvas. Type the answers for #1 and #9 at the end of the document. 1.
We then obsevered the two slides for number of cells as well as for food vacuoles inside a cell using a microscope at times of 0,5,10,20, and 30 minutes. Results The following graphs show the results of this experiment. The tetrahymena sample that was introduced to concentrated tobacco had a lower cell/vacuole ratio than the tetrahymena sample that was not exposed to
In this lab when looking at cells, we observed the salinity and osmolarity of the cell when placed in the environment. With the different concentrations of NaCl, we are able to see how different environment can constrain an organism and see the wide range of responses to regulate in cell’s osmolarity. The cells we studied was sheep red blood cells (erythrocyte), because they are the most studied membrane system and therefore used as ideal membrane to study the relationship between water and the passing of the different concentration of NaCl across the membrane. The purpose of the experiment was to observe the cell and infer which direction of the flow of the water due to the cell volume change.
This root tip was choosen because of its rapid growth and it can be easily avaliable and grown in large numbers. The rapid root growth proved advantageous as it allowed the observation of multiple cells in each mitotic stage within a small sample. It was expected that the majority of the cells found would be in interphase as a large proportion of the cell division cycle is spent with the cell performing its normal cellular functions. Materials: The Materials required for this experiment include; a
Additionally, mitotic spindle disintegrates and chromosomes are uncoiled into chromatin, returning the cell to the interphase structure. Even though mitosis is finished with telophase, the final cell segregation occurs at cytokinesis. It starts before telophase and divides the cytoplasm of the cell, therefore making two daughter cells with the genetically identical nucleus. The aim of the practical was to observe the sample of the onion root tip under the microscope and to count cells in different mitotic phases.
Demonstration of Cellular Respiration of a celery cell in vivo. Purpose Cellular respiration is a metabolic process consisting of a series of oxidation reactions in which oxygen is utilized and therefore is called an aerobic reaction. The process of cellular respiration takes place in the mitochondria, located in the cell of an organism and which converts biochemical energy from nutrients into adenosine triphosphate (ATP), and then releases waste products. The mitochondria is surrounded by two membranes, the inner membrane and outer membrane.
→ Calculation of the absolute values of the different leukocytes in the differential was accomplished via the following formula: (total WBC count x relative value in %)/100 On the chemistry tests: increased uric acid (8.4 with normal being 4-6 mg/dL) increased lactate dehydrogenase (LD) (692 with normal being 140-280
However, cancer treatments nowadays are very risky and have severe side effects such as blood clotting, bone issues, and hair loss. More efforts should be put in the deeper understanding of glycolysis and other pathway. In addition, it would me more efficient to find other treatments with less side effects that target the stage of glycolysis in tumor cells, inhibiting the overexpressing of enzymes and the production of lactic, leading to stop the abnormal cell division, and cure
They are not exposed to as many outside factors as epithelial cells, in skin cells, therefore not dividing as much as when needed. The Allium Root Tip cell average mitotic index, (the number of cells going through mitosis compared to the total number of cells), was greater than human bone cells. The way different cells react to mitosis, and stage length