Ashley Renner
Martin Luther King Jr. Magnet
AP Biology, Biology Teacher

Biomedical Optics Laboratory

Josh Beckham, Ph.D. Candidate

I worked in a Biomedical Optics Laboratory under the supervision of Josh Beckham.  There, my research focused on determining collateral tissue damage caused by lasers.  The cell line that I worked with was derived from a mouse embryonic epithelial cell, specifically NIH3T3.  The cell line was transfected to produce two cell types, pGL3 and HSP70luc.  Cells of the pGL3 line were transfected with the pGL3 plasmid that allowed constant expression of a gene called luciferase.  Cells of the HSP70luc (Heat Shock Protein 70) were transfected to incorporate the luciferase gene with the HSP Promoter.  Cells of this line required heat to cause the HSP 70 promoter to produce luciferase.              

The goal of my research was to determine amounts of tissue damage by exposing the two cell lines to varying amounts of heat energies and time exposures.  To accomplish this, a HoYAG laser of 488nm was used to lase the cells.  A computer program was developed to lase a specific pattern on our cell plates.  After laser exposure the cells were incubated then treated with luciferin.  The luciferin reacted directly with the luciferase gene causing luminescence.

Cells of the pGL3 line luminesced if they were alive and received no tissue damage.  Cells that were damaged or killed were unable to produce luciferase and therefore a reaction with luciferin was unable to take place.  Cells of the HSP70 line luminesced if they received adequate heat shock to produce luciferase.  See images below. 

pGL3 Cells                                                      HSP70 Cells

Other tasks of my research included cell culturing, Cell Titer Blue Assays, gel electrophoresis, cell viability tests, and DNA analysis of a particular Heat Shock Protein gene.

I also had the opportunity to tour facilities and learn about equipment that were relevant to my research experience.      

My research ultimately focused on genetic engineering and transformation.  The curriculum encourages students to develop genetic engineering skills and learn about current research for genetic diseases such as Cystic Fibrosis, while solving inquiry-based challenges.