My internship at the weiss lab

1. Fall 2015
Amelia Payne | Big Picture South Burlington

2. Introduction
• Feel free to use electronic devices during my presentation
• Ask questions during presentation
• You will get candy

3. Introduction
• Internship
• Classes
• Online Learning
• Time tracking
• Clubs
• STP

4. Internship At
Weiss Lab

5. Freezing Decellurized Lungs
• Cultivate cells in sterile conditions
• Freeze tissue in different mediums
• Defrost tissues and seed with cells
• Test for cell growth with WST-assay

6. Growing Cell Lines
Human Lung Fibroblasts (HLF)
• Defrosted cells
• Grew under sterile conditions
• Counted
• Replated and frozen

7. Counting
• Inject 10 µl of cell suspension into “Neubauer chamber”
(device for cell counting)
• Count number of cells in one quadrant
• Multiply by 10^4 to get total cells per ml cell suspension

8. Freezing
Lungs
• At rate of 1°C per minute
to -80°C
• Transferred to liquid
nitrogen (-195°C = -320°F)
for long term storage
• Will defrost at 1, 3, 6, and
12 months and do WST-
assay
• Control: fresh tissue
• We froze tissue in
• 5% glycerol
• 10 % DMSO in PBS

9. Seeding
Defrosted
Tissue
We will defrost tissue
rapidly and seed with
cells once we reach the
first time points.

10. WST-assay
• Indicates metabolic activity of
cells
• WST-1 (water soluble
tertrazolium salt-1, red) is
cleaved into formazan (yellow)
by an enzyme produced by the
mitochondria. The resultant
color changes indicates
metabolic activity of the cells
• Absorbance measurement
(440 nm) to determine how
much formazan was produces

11. Data from WST Assay Control

12. The Team
Thanks to all my awesome
mentors who are here today
*Sorry if you’re not in the
picture because you weren't
at lunch on Thursday.