Eukaryotic Cells

1. Cell Culture

   1. Introduction to cell culture

   2. Media preparation

   3. DMEM

   4. Dialysis of FBS

   5. Splitting

   6. Hemacytometer

   7. Coulter counter

   8. Trypan blue staining

   9. Freeze/thaw

   10. Freezing cell lines

   11. Thawing cells

   12. HeLa S3 cell culture

   13. Xenopus oocyte microinjection

   14. Preparation of extracts from human placenta

   15. Chicken blood injections

   16. Sacrifice of adult hen

   17. Bleeding eggs

   18. Chicken embryonic fibroblasts

2. Cell Transfection

   1. CaPO4 method

   2. 3% CO2 method

   3. Ecogpt

   4. Trypsinization transfection

   5. Polyfect, Lipofectamine, and Effectene

   6. Cell sorting for transfected cells: MACs protocol

3. Establishing Stable Cell Lines

   1. Calcium Chloride Transfection

   2. Lipofectamine Transfection (Gen’s Protocol)

4. Radiolabeling

   1. 32P orthophosphate and 35S methionine/cysteine

   2. Metabolic labeling

5. Cell Growth/Death Assays

   1. Cell Death ELISA

   2. MTT

   3. TUNEL

   4. Drug Assays

6. Luciferase Assays

   1. Luminometer operation

   2. Single luciferase

   3. Dual luciferase

   4. 96-well plate format

7. RNAi – mammalian cells

8. Cell Imaging

   1. Intro to immunofluorescence

   2. Method

   3. FRET

9. Transferrin Uptake using Fluorescence Microscopy