Cell Biology Lab Manual

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Mammalian Cell Culture Lab Manual
Student guide of Cell Biology Laboratory Techniques

2012
Jay M. Feldman
Cell Biology Professor Lagier
11/26/2012
Mammalian Cell Culture Lab Manual
Student guide of Cell Biology Laboratory Techniques

2012
Jay M. Feldman
Cell Biology Professor Lagier
11/26/2012

Table of Contents
I. Introduction
* About the authorpage 2
* Prefacepage 3

II. Laboratory Techniques
* Cell Culture Techniquepage 4-5
* Sterile Techniquepage 6-7
* Media Preparationpage 8-9
* Seed cellspage 10
* Subculturepage 11-12
* Cell countpage 13-14
* Cryopreservationpage 15

III. Laboratory Experiments
* Intentional Contaminationpage 16-17
* Tonicitypage 18-19
* DNA Transfectionpage 20-23
* Cytoskeletonpage 24-25

IV.
* Answer Keypage 26-27
* Works Citedpage 28

About the Author
Hi there! My name is Jay Feldman and I am a junior at Florida Gulf Coast University. I am biology major with a concentration in pre-professional studies. As a 20 year-old full time student, I also work part-time in the emergency room at NCH North Collier. My job title is an emergency department scribe and I have held this position for one year. In my free time I enjoy weight lifting, basketball, and partying with my friends. I am a proud member of Sigma Phi Epsilon fraternity as well as the beach volleyball club and the men’s indoor volleyball team at FGCU. My goal in life is to become a big time plastic surgeon in the city, I love medicine and I work hard every day to achieve my goals. I hope you enjoy my lab manual, be nerdy!

Preface

This laboratory manual was created in order to direct future students through to various laboratory procedures and concepts that a student will learn throughout a semester of cell biology. Following the guidelines in this lab manual will direct you in the techniques to carry out procedures, keep track of your results, and determine if your experiment was a success. This manual will only cover labs having to do with the eukaryotic cell, which was the only cell that was studied throughout the course of the semester. This manual will cover a general list of cell culture techniques, a description and function of some general lab equipment, and various lab procedures. These lab assignments include the intentional contamination lab, Tonicity lab, DNA transfection, Cytoskeleton lab, as well as how to prepare your media, how to count cells, and how to use sterile technique in your lab experiments. This laboratory manual will also include several questions that are included in order to test your understanding of the material that is covered throughout the script. I suggest that you answer the questions at the end of each section in order to strengthen your understanding of cell biology lab techniques. I hope that you enjoy, be nerdy!

Introduction to Cell Culture Techniques

* What is Cell Culture?

Cell culture (tissue culture) is the process of growing cells in an artificial environment outside of the organism, also called (in vitro)

-The artificial environment we will be using is called ‘media’

* Advantages and disadvantages of growing cells in culture?

Advantages:
1) Ability to control the physiochemical environment (pH, temperature, oxygen, carbon dioxide, osmotic pressure, etc.) 2) Controlled and defined physiological conditions (constitution of medium, etc.) Disadvantages:

1) More expensive
2) Requirement of controlled and defined physiological conditions

* Equipment/Supplies

1) Biology safety cabinet- Class II: Hood used for cell culture; protects environment, person, and the cell line from contamination from outside environment 2) CO2 Incubator: Environment where cells are grown; maintains cells original environment by keeping temperature at 37 degrees Celsius, CO2 levels at 5%, and maintaining a humidity...
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