UMBC Institute of Fluorescence (IoF)

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http://hdl.handle.net/11603/10906

The IoF was founded in 2001 by Dr. Chris D. Geddes, Professor and Director. Under the leadership of Dr. Chris D. Geddes, the IoF has earned a well-deserved international reputation for its advances in Fluorescence Spectroscopy and Plasmonics. Approaches and concepts both developed and discovered by the group, such as Metal-Enhanced Fluorescence (MEF), Metal-Enhanced Chemiluminescence (MEC), Surface-Plasmon Coupled Phenomenon and the glucose-sensing contact lens, are both well-recognized, highly cited and currently used in laboratories around the world today.

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Now showing 1 - 11 of 11

Heavy carbon nanodots: a new phosphorescent carbon nanostructure
(Royal Society of Chemistry, 2018-05-16) Knoblauch, Rachael; Bui, Brian; Raza, Ammar; Geddes, Chris
Carbon nanodots are nanometer sized fluorescent particles studied for their distinct photoluminescent properties and biocompatibility. Although extensive literature reports the modification and application of carbon nanodot fluorescence, little has been published pertaining to phosphorescence emission from carbon nanodots. The use of phosphors in biological imaging can lead to clearer detection, as the long lifetimes of phosphorescent emission permit off-gated collection that avoids noise from biological autofluorescence. Carbon nanodots present a desirable scaffold for this application, with advantageous qualities ranging from photostability to multi-color emission. This research reports the generation of a novel phosphorescent “heavy carbon” nanodot via halogenation of the carbon nanodot structure. By employing a collection pathway that effectively incorporates bromine into the nanostructure, T₁ triplet character is introduced, and subsequently phosphorescence is observed in liquid media at room temperature for the first time in the nanodot literature. Further experiments are reported characterizing the conditions of observed phosphorescence and its pH-dependence. Our approach for producing “heavy carbon nanodots” is a low-cost and relatively simple method for generating the phosphorescent nanodots, which sets the foundation for its potential future use as a phosphorescent probe in application.
Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
(PLOS, 2018-07-25) Santaus, Tonya; Li, Shan; Ladd, Paula; Harvey, Amanda; Cole, Shannon; Stine, O. Colin; Geddes, Chris
Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It® for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It® employs a conventional microwave and Lyse-It® slides for intensely focused microwave irradiation onto the sample. High microwave power and a <60 second irradiation time allow for rapid cellular lysis and subsequent intracellular component release. The pathogenic bacteria are identified by quantitative polymerase chain reaction (qPCR), which subsequently demonstrates the viability of DNA for amplification post microwave-induced lysis. Intracellular component release, degradation, and detection of L. monocytogenes and V. cholerae has been performed and shown in this paper. These results demonstrate a rapid, low-cost, and efficient way for bacterial sample preparation on both food and water-borne Gram-positive and -negative organisms alike.
Molecular Characterization of Markers Associated With Antimicrobial Resistance in Neisseria gonorrhoeae Identified From Residual Clinical Samples
(American Sexually Transmitted Diseases Association, 2018-05) Melendez, Johan; Hardick, Justin; Barnes, Mathilda; Barnes, Perry; Geddes, Chris; Gaydos, Charlotte A.
Background The emergence and spread of antimicrobial-resistant (AMR) Neisseria gonorrhoeae (NG) is a major public health concern. In the era of nucleic acid amplifications tests, rapid and accurate molecular approaches are needed to help increase surveillance, guide antimicrobial stewardship, and prevent outbreaks. Methods Residual urethral swabs, collected prospectively in the Baltimore City Health Department during a 6-month period, were analyzed by real-time polymerase chain reaction assays for NG DNA and AMR determinants to fluoroquinolones, penicillin, and extended-spectrum cephalosporins. Results N. gonorrhoeae DNA was detected in 34.8% (73/210) of samples, including 67.3% (68/101) of the swabs that had been previously identified as NG positive by culture. Markers associated with decreased susceptibility to fluoroquinolones were detected in 22.4% of the polymerase chain reaction NG-positive samples. The rate of penicillinase-producing NG was very low (1.6%), and no markers associated with decreased susceptibility to extended-spectrum cephalosporins were detected in this cohort of men using the AMR assays herein described. Conclusions Detection of molecular markers associated with AMR in NG can be performed directly from residual clinical samples, although the recovery rate of adequate DNA for molecular testing from these samples can be suboptimal. A high number of samples with mutations associated with decreased susceptibility to fluoroquinolones were identified.
Heavy carbon nanodots 2: plasmon amplification in Quanta Plate™ wells and the correlation with the synchronous scattering spectrum
(Royal Society of Chemistry, 2018-12-14) Knoblauch, Rachael; Ra, Estelle; Geddes, Chris
Brominated carbon nanodots are a new carbon nanostructure that exhibits strong phosphorescence without fixation. Herein we report plasmonic amplification of this phosphorescence in silver-coated Quanta Plate™ wells, a technique called metal-enhanced phosphorescence (MEP). Subsequently we correlate the excitation and emission components of brominated carbon nanodots to their respective enhancement values. These properties are then discussed in relation to the synchronous scattering spectrum of the plasmonic substrate, in the first report of its kind for MEP. These results set the foundation for expanded application of carbon nanodots, as the photophysical characteristics of phosphorescence are improved, and augment the growing understanding of MEP.
Spectral Distortions in Metal-Enhanced Fluorescence: Experimental Evidence for Ultra-Fast and Slow Transitions
(ACS Publications, 2020-02-05) Knoblauch, Rachael; Hamo, Hilla Ben; Marks, Robert; Geddes, Chris
Metal-enhanced fluorescence (MEF) has become an increasingly important technology in recent years, with thorough research addressing the fundamentals of MEF. In many studies, spectral distortion is observed in the enhanced spectra as compared to free-space fluorescence emission profiles. Despite this observation, very little experimentation has hitherto been undertaken to investigate the mechanistic underpinnings of spectral distortion in MEF. Herein, we investigate MEF spectral distortion using Rose Bengal and fluorescein on silver nanoparticle substrates, subsequently isolating the coupled fluorescence spectrum for a deeper understanding of the spectral modifications. Clear experimental evidence for bathochromic distortion is reported. Remarkably, we also report hypsochromic distortion in one of the first experimental observations of plasmonic coupling to high-energy excited states. Additionally, the coupled fluorescence spectra from other published literature have also been both extracted and examined, and the subsequent spectral distortions are reported here. The previously asserted theory of radiative decay rate modification for spectral distortion is discussed in the context of both plasmonic properties as well as fluorophore photophysical characteristics including lifetime and quantum yield. The dual enhancement mechanism of MEF is also explored in the context of spectral distortion. The results and discussion reported herein subsequently provide one of the first comprehensive examinations of spectral distortion in MEF to date.
Spectral distortions in zinc-based metal-enhanced fluorescence underpinned by fast and slow electronic transitions
(Elsevier, 2020-02-19) Knoblauch, Rachael; Hamo, Hilla Ben; Marks, Robert; Geddes, Chris
Metal-enhanced fluorescence (MEF) is a promising technology with impact in diagnostics, electronics, and sensing. Despite investigation into MEF fundamentals, some properties remain unresearched, notably spectral distortion. To date, publications have described its underpinnings, yet comprehensive analysis is needed, as presented recently for silver films. Herein we expand this description using zinc substrates (ZnNPs). Significant red-edge and blue-edge distortions are reported using Rose Bengal. Radiative decay rate modification is identified as key in amplifying fast/slow electronic transitions by the enhanced emission mechanism. Furthermore, we identify distortion in published studies, bolstering our thinking that spectral distortion is an intrinsic property of MEF.
Antimicrobial carbon nanodots: photodynamic inactivation and dark antimicrobial effects on bacteria by brominated carbon nanodots
(Royal Society of Chemistry, 2020-11-11) Knoblauch, Rachael; Harvey, Amanda; Ra, Estelle; Greenberg, Ken M.; Lau, Judy; Hawkins, Elizabeth M.; Geddes, Chris
The evolving threat of antibiotic resistance development in pathogenic bacteria necessitates the continued cultivation of new technologies and agents to mitigate associated negative health impacts globally. It is no surprise that infection prevention and control are cited by the Centers for Disease Control and Prevention (CDC) as two routes for combating this dangerous trend. One technology that has gained great research interest is antimicrobial photodynamic inactivation of bacteria, or APDI. This technique permits controllable activation of antimicrobial effects by combining specific light excitation with the photodynamic properties of a photosensitizer; when activated, the photosensitizer generates reactive oxygen species (ROS) from molecular oxygen via either a type I (electron transfer) or type II (energy transfer) pathway. These species subsequently inflict oxidative damage on nearby bacteria, resulting in suppressed growth and cell death. To date, small molecule photosensitizers have been developed, yet the scalability of these as widespread sterilization agents is limited due to complex and costly synthetic procedures. Herein we report the use of brominated carbon nanodots (BrCND) as new photosensitizers for APDI. These combustion byproducts are easily and inexpensively collected; incorporation of bromine into the nanodot permits photosensitization effects that are not inherent to the carbon nanodot structure alone—a consequence of triplet character gained by the heavy atom effect. BrCND demonstrate both type I and type II photosensitization under UV-A irradiation, and furthermore are shown to have significant antimicrobial effects against both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus and Listeria monocytogenes as well. A mechanism of “dark” toxicity is additionally reported; the pH-triggered release of reactive nitrogen species is detected from a carbon nanodot structure for the first time. The results described present the BrCND structure as a competitive new antimicrobial agent for controllable sterilization of bacteria.
Development of a Microplate Platform for High-Throughput Sample Preparation Based on Microwave Metasurfaces
(IEEE, 2021-03-08) Nichols, Zach E.; Saha, Lahari; Knoblauch, Rachael; Santaus, Tonya; Geddes, Chris
Sample preparation is one of the most time-consuming steps in diagnostic assays, particularly those involving biological samples. In this paper we report the results of finite-difference time-domain (FDTD) simulations and thermographic imaging experiments carried out with the intent of designing a microplate for rapid, high-throughput sample preparation to aid diagnostic assays. This work is based on devices known as microwave lysing triangles (MLTs) that have been proven capable of rapid sample preparation when irradiated in a standard microwave cavity. FDTD software was used to model a microplate platform as a polystyrene substrate with an array of various passive scattering elements (PSEs) of different sizes, shapes, and interelement spacings in a 2.45 GHz field identical to that of a common microwave oven. Based on the FDTD modeling, several PSE arrays were fabricated by cutting PSEs out of aluminum foil and adhering them to the bottom of regular polystyrene microplates to make prototypes. Each prototype microplate was then irradiated in a microwave cavity for a range of different times, powers, and source angles and the heating effects were observed via a forward looking infrared (FLIR) camera. Based on the results, two prototype microplate platforms have been shown to demonstrate electromagnetic and thermal enhancements similar to those seen with MLTs as well as tunable thermal responses to radio frequency (RF) irradiation.
Elucidation of a non-thermal mechanism for DNA/RNA fragmentation and protein degradation when using Lyse-It
(PLOS, 2019-12-02) Santaus, Tonya M.; Greenberg, Ken; Suri, Prabhdeep; Geddes, Chris D.
Rapid sample preparation is one of the leading bottlenecks to low-cost and efficient sample component detection. To overcome this setback, a technology known as Lyse-It has been developed to rapidly (less than 60 seconds) lyse Gram-positive and–negative bacteria alike, while simultaneously fragmenting DNA/RNA and proteins into tunable sizes. This technology has been used with a variety of organisms, but the underlying mechanism behind how the technology actually works to fragment DNA/RNA and proteins has hitherto been studied. It is generally understood how temperature affects cellular lysing, but for DNA/RNA and protein degradation, the temperature and amount of energy introduced by microwave irradiation of the sample, cannot explain the degradation of the biomolecules to the extent that was being observed. Thus, an investigation into the microwave generation of reactive oxygen species, in particular singlet oxygen, hydroxyl radicals, and superoxide anion radicals, was undertaken. Herein, we probe one aspect, the generation of reactive oxygen species (ROS), which is thought to contribute to a non-thermal mechanism behind biomolecule fragmentation with the Lyse-It technology. By utilizing off/on (Photoinduced electron transfer) PET fluorescent-based probes highly specific for reactive oxygen species, it was found that as oxygen concentration in the sample and/or microwave irradiation power increases, more reactive oxygen species are generated and ultimately, more oxidation and biomolecule fragmentation occurs within the microwave cavity.
Effects of Lyse-It on endonuclease fragmentation, function and activity
(PLOS, 2019-09-30) Santaus, Tonya M.; Zhang, Fan; Li, Shan; Stine, O. Colin; Geddes, Chris D.
Nucleases are enzymes that can degrade genomic DNA and RNA that decrease the accuracy of quantitative measures of those nucleic acids. Here, we study conventional heating, standard microwave irradiation, and Lyse-It, a microwave-based lysing technology, for the potential to fragment and inactivate DNA and RNA endonucleases. Lyse-It employs the use of highly focused microwave irradiation to the sample ultimately fragmenting and inactivating RNase A, RNase B, and DNase I. Nuclease size and fragmentation were determined visually and quantitatively by SDS polyacrylamide gel electrophoresis and the mini-gel Agilent 2100 Bioanalyzer system, with a weighted size calculated to depict the wide range of nuclease fragmentation. Enzyme activity assays were conducted, and the rates were calculated to determine the effect of various lysing conditions on each of the nucleases. The results shown in this paper clearly demonstrate that Lyse-It is a rapid and highly efficient way to degrade and inactivate nucleases so that nucleic acids can be retained for down-stream detection.
A comparison of Lyse-It to other cellular sample preparation, bacterial lysing, and DNA fragmentation technologies
(PLOS, 2019-07-23) Santaus, Tonya M.; Li, Shan; Saha, Lahari; Chen, Wilbur H.; Bhagat, Siya; Stine, O. Colin; Geddes, Chris D.
The ability for safe and rapid pathogenic sample transportation and subsequent detection is an increasing challenge throughout the world. Herein, we describe and use bead-beating, vortex, sonication, 903 protein saver cards, and Lyse-It methods, aiming to inactivate Grampositive and -negative bacteria with subsequent genome DNA (quantitative Polymerase Chain Reaction) qPCR detection. The basic concepts behind the four chosen technologies is their versatility, cost, and ease of use in developed and underdeveloped countries. The four methods target the testing of bacterial resilience, cellular extraction from general and complex media and subsequent DNA extraction for qPCR detection and amplification. These results demonstrate that conventional high temperature heating, 903 protein saver cards, and Lyse-It are all viable options for inactivating bacterial growth for safe shipping. Additionally, Lyse-It was found to be particularly useful as this technology can inactivate bacteria, extract cells from 903 protein saver cards, lyse bacterial cells, and additionally keep genomic DNA viable for qPCR detection.

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