Maximum diversity of life exists within the estuaries and coral reefs of the Globe. The absence of vertebrate and other land dwelling adaptations has resulted in an enormous range of complexity among invertebrates and their symbiotic biome resulting in the generation of compounds finding uses in anti-tumor and antibiotic applications. It has been widely reported that the greatest factor limiting progress in characterizing and processing new therapeutics derived from invertebrates is the lack of adequate original material. Symbiotic bacteria within specific tunicates often synthesize antitumor compounds as secondary metabolites. We describe a 3-stage protocol that utilizes acoustic and photonic analysis of large areas of marine ecosystem and life forms. We refer to this as Estuary Assessment System (EAS), which includes a multi-frequency acoustic transducer/sensing instrument mounted on our research vessel. This generates a topological map of surveyed tracks of marine locations known to be habitats of useful actinobacteria laden invertebrates. Photonic devices are used to generate image and pulse data leading to location, identification and isolation of tunicates and actinobacteria.
Highly evolved bacteria living within immobile marine animals are being targeted as a source of antitumor pharmaceuticals. This paper describes 2 electo-optical sensor systems developed for identifying species of tunicates and actinobacteria that live within them. Two stages of identification include 1) a benthic survey apparatus to locate species and 2) a laboratory housed cell analysis platform used to classify their bacterial micro-biome. Marine Optics Sampling- There are over 3000 species of Tunicates that thrive in diverse habitats. We use a system of cameras, GPS and the GPS/photo integration application on a PC laptop to compile a time / location stamp for each image taken during the dive survey. A shape-map of x/y coordinates of photos are stored for later identification and sampling. Flow Cytometers/cell sorters housed at The Medical University of South Carolina and The University of Maryland have been modified to produce low-noise, high signal wave forms used for bacteria analysis. We strive to describe salient contrasts between these two fundamentally different sensor systems. Accents are placed on analog transducers and initial step sensing systems and output.
Spectroscopic and light scattering methods were used to gain insight into the existence
and characterization of the cancer stem cell. Fundamental technical description of devices
used have been reported elsewhere. We included alterations and implementation of these
biophotonic instruments as applied to our objectives. We disassociated human tumor and
submitted the cells to optical characterization to support our working hypothesis of stem
cell origins to cancer and mechanisms. Single cell combined with population based
analysis within the Pancreatic cancer system led us to information regarding the
polarization state of cells possessing anchor proteins and drug influx pumps.
Multispectral imaging combined with flow cytometry enabled us to target rare cells that
appear to retain template DNA. rendering them resistant to anti-cancer drug therapy. In
this study we describe an optical method that combines high-throughput population
pattern and correlates each cell with an individual fluorescent and bright-field image.
We designed and fabricated a flow cytometry work bench for the purpose of testing optical schemes utilized in
two specialized flow cytometry techniques. Fluorescent spectroscopy and multi-angle scattering of laser light
has potential for generating novel classes of cell population information. The acquisition of full fluorescence
spectrum of dye bound to living cells may associate specific function to spectral shift. Similarly, detection of 2
discrete angles of light scatter was shown to describe internal structures of cells based on refractive and
diffractive properties. We utilized a flow cytometry electro- cells based on refractive and diffractive properties.
We utilized a flow cytometry electro- optical test bench to develop and describe alternate light collection
schemes for these two techniques. Mouse bone marrow was chosen as the subject of analysis because it contains
a consistent mix of cells that possess distinct biophysical properties. More importantly, these cells are the source
of various biological systems that are targeted by environmental stress and clinical disease.