Illuminating Biology from Benchtop to Bedside: Biophotonics across Energy, Space, and Time
Biophotonics refers to the use of light to image, probe, manipulate or modify biological systems. The use of biophotonics methods has become increasingly pervasive throughout the biomedical sciences. We present current research from our IGERT trainees that highlights the utility of biophotonics across energy, space, and time to address a diversity of biomedical issues. In one project we are developing a non-linear imaging system capable of generating sub-micron resolution, 3D images of biological structures within a minute. This device will be used to study collagen fiber nanostructure. A second project utilizes highly-focused laser pulses to generate microscale cavitation bubbles to study cellular mechanotransduction with potential applications to drug screening. In a third research project we are developing bioluminescent probes using Gaussia luciferase that will provide dynamic information regarding cell-cell interactions. This technology will be used to generate images of tumor cell interaction on the micrometer scale within a minute.
In addition to basic science investigations, our IGERT group is also developing biophotonics technologies for clinical translation. We are developing a long-range optical coherence tomography system capable of generating 3D images at 50 frames per second with a resolution of 10 microns, which will be used in the clinic to evaluate sleep apnea. A second clinical translation project focuses on the development of spatial frequency domain imaging, which provides wide-field functional images of biochemical tissue compositions on the order of a minute with millimeter resolution. SFDI will be used in the clinic to monitor kidney health during partial nephrectomy procedures.
Jon Kellar
Faculty: Project Co-PI
What is the mechanism by which HUVECs release Ca++ from the endoplasmic reticulum?
Justin Luo
Graduate Student
Hello Professor Kellar,
The cavitation bubble will expose HUVECs to mechanical deformation and stimulate G-protein coupled receptors on the cell membrane. Stimulation of the G-protein coupled receptor will lead to production of inositol triphosphate (IP3). IP3 is produced from the protein phospholipase C (PLC) cleaving phosphatidylinositol 4,5-biphosphate (PIP2). IP3 diffuses and binds to the IP3 receptor on the endoplasmic reticulum surface which then induces Ca++ release.
Thank you for your question.
Sincerely,
Justin
Marc Porter
Faculty: Project PI
What is the specific structure-function relationship(s)you are attempting to examine the cornea project? That is, what are the minor changes (not clear what you mean by “arrangement”) in the corneal lamella you are attempting to connect to cornea function?
Julie Hsu
Graduate Student
Hello Dr. Porter,
Structurally, type I collagen is known to be composed of fibrils which align in a parallel or antiparallel fashion to produce collagen fibers and lamellae. It has been observed through methods such as electron microscopy and second harmonic generation how the collagen lamellae orient themselves in three-dimensional space within the cornea, forming layers which run in different directions depending on the region within the cornea. Fibers of one lamella may interweave with adjacent lamellae, forming a pattern that is believed to provide structural support for the cornea as well as functional importance. However, although methods such as these can image the physical orientation of lamellae and the fibers within them, they are unable to provide information on the polarity of the molecules. Therefore, one can not distinguish between parallel and antiparallel fibers within lamellae. The sum frequency generation microscope that we use has the capability to resolve these structural questions through implementation of an interferometric setup providing phase-sensitive detection and enabling polarization studies. It is of interest to us whether more understanding of how the cornea functions can be obtained from studying the polarity orientations of fibers within lamellae, particularly those that participate in interweaving.
Thank you for your question.
Sincerely,
Julie
Adriane Ludwick
Faculty: Project Co-PI
For any of the five presenters…or all…how would this approach apply to a study of the various heart-related illnesses?
Joe Jing
Graduate Student
Hello Dr. Ludwick,
Cardiovascular studies are a very popular field within biophotonics. Fiber optic technologies allow for the delivery of light to many locations including the heart. Our group is actually actively studying cardiovascular plaques using Optical Coherence Tomography. We have built miniature fiber probes that can be inserted from the femoral artery and guided to lesions of interest for precise imaging. We can then measure different properties such as the elastic properties and wall thickness to better assess possibly plaque rupture. Multiphoton imaging can further improve upon these diagnostics as well by specifically targeting for molecular contrast such as lipids which form the interior of plaques. With these techniques, we can asses plaques at far higher resolutions and with higher specificity than compared to current techniques such as intravascular ultrasound.
Thank you for your question,
Joe
Peter Gannett
Faculty: Project Co-PI
Are there potential applications for the IRI technology for stroke, either for basic research or clinical applications?
Kyle Nadeau
Hi Dr. Gannett,
Thanks for your question. We are in fact interested in monitoring stroke using our technology, and have published work pertaining to a preclinical rat model for stroke. Since most strokes are a result of ischemia, we can monitor this process in a similar fashion to how we monitor IRI shown in our presentation. In our stroke model, we perform an open vascular occlusion on the middle cerebral artery. Using SFDI, we can monitor light absorption and scattering in the rat brain. By interrogating the brain at several wavelengths of light, we are able to derive values for oxy/deoxy hemoglobin, from which we can quantify oxygen saturation, and thus tissue metabolism. In a similar manner to our kidney IRI study, we can also use light scattering changes to infer micro-scale changes such as cellular swelling and tissue edema.
Sincerely,
Kyle Nadeau
Antal Jakli
Faculty: Project Co-PI
Can you explain how non-centrosymmetry leads to a sum-frequency generation?
Julie Hsu
Graduate Student
Hi Dr. Jakli,
When a sample is driven by waves of frequency w1 and w2, the molecules in the sample act like anharmonic oscillators, producing oscillations at frequency w1+w2. The induced dipole can be expressed as P(w1+w2)=X*E(w1)E(w2), X being the nonlinear susceptibility. For a centrosymmetric sample that has inversion symmetry, if the sample were driven by -E(w1) and -E(w2) instead, the induced dipole should be -P(w1+w2). However, the equation shows us that X-E(w1)*-E(w2) still produces P(w1+w2). In order for this to be true, X must be zero. Therefore, the sum frequency generation only exists for samples with non-centrosymmetry.
Thank you for your question.
Sincerely,
Julie