Devices, methods, and systems for fluorescence-based imaging and detection of location(s) and/or source(s) of potential contamination and/or pollution
A method for fluorescence-based imaging of a target to detect contamination and/or pollutants is disclosed. The method includes labelling a pre-selected biomarker at the target, illuminating the target with excitation light emitted by an excitation light source and having at least one wavelength or wavelength band causing at least the pre-selected biomarker to fluoresce, detecting fluorescence emissions of at least the pre-selected biomarker with an image detector of a handheld imaging device, and determining the presence, location, and/or quantity of contamination and/or pollutants on and/or in the illuminated target based on the detected fluorescence emissions of at least the pre-selected biomarker.
1 . A method for fluorescence-based imaging of a target to detect biological contamination of the target, comprising:
applying at least one fluorescing contrast agent to a target, wherein the at least one fluorescing contrast agent includes an exogenous, bacteria-specific contrast agent and is configured to label one or more pre-selected biomarkers at the target;
subsequent to application of the at least one fluorescing contrast agent, illuminating the target with excitation light emitted by an excitation light source of a handheld imaging device and having at least one wavelength or wavelength band configured to cause the one or more pre-selected biomarkers to fluoresce;
detecting fluorescence emissions from the one or more pre-selected biomarkers with an image detector of the handheld imaging device;
illuminating the target with white light emitted by a white light source of the handheld imaging device;
detecting reflections from the target in response to illumination of the target with the white light;
comparing fluorescence emission band(s) of the one or more pre-selected biomarkers to a predetermined look-up table of fluorescence emission spectra or fluorescence emission band(s) of biomarkers;
identifying one or more of a presence, a location, and a quantity of biological contamination on or in the target based, at least in part, on the detected fluorescence emissions of the one or more pre-selected biomarkers, wherein the biological contamination comprises one or more of colonies of bacteria and biological fluids; and
displaying a representation of the target indicative of one or more of the presence, the location, and the quantity of the biological contamination on or in the target, wherein the representation of the target spatially co-registers fluorescence emission data from the detected fluorescence emissions of the one or more pre-selected biomarkers and reflection data from the target and comprises fluorescence red-blue-green (RBG) ratio data displayed in combination with reflectance RBG ratio data.
2 . The method of claim 1 , wherein applying the at least one fluorescing contrast agent to the target comprises applying a combination of two or more contrast agents to the target, wherein the combination is specific to the one or more pre-selected biomarkers.
3 . The method as claimed in claim 1 , wherein the exogenous, bacteria-specific contrast agent is configured to make visible biological contamination comprising colonies of one or more of Listeria monocytogenes, Enterobacter sakazakii, Campylobacter coli, Campylobacter jejuni, Campylobacter lari , coliform bacteria, bacteria of the E. coli species, Salmonella , bacteria of the Staphylococcus aureus species, bacteria of the Staphylococcus genus, and Pseudomonas aeruginosa in or on the target.
4 . The method of claim 1 , wherein identifying one or more of the presence, the location, and the quantity of the biological contamination on or in the target includes detecting one or more of a presence, a location, and a quantity of a bacterial strain based on the fluorescence emissions of the one or more pre-selected biomarkers.
5 . The method as claimed in claim 4 , wherein the bacterial strain is at least one selected from the group consisting of Staphylococcus bacteria, Staphylococcus aureus, Pseudomonas aeruginosa, Listeria monocytogenes, Enterobacter sakazakii, Campylobacter species of bacteria, coliform bacteria, Escherichia coli bacteria, Propionibacterium acnes and Salmonella.
6 . The method of claim 4 , wherein detecting the one or more of the presence, the location, and the quantity of the bacterial strain based on the fluorescence emissions of the one or more pre-selected biomarkers includes differentiating one or more of the presence, the location, and the quantity of two or more different bacterial strains based on the fluorescence emissions of the one or more pre-selected biomarkers.
7 . The method of claim 6 , wherein the different bacterial strains include Staphylococcus aureus and Pseudomonas aeruginosa , and the different bacterial strains are differentiated based on their respective fluorescence emission signatures.
8 . The method of claim 1 , wherein the one or more pre-selected biomarkers are chosen from the group consisting of: bacteria, fungi, yeast, spores, virus, microbes, parasites, connective tissues, tissue components, exudates, pH, blood vessels, reduced nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), microorganisms, vascular endothelial growth factor (VEGF), endothelial growth factor (EGF), epithelial growth factor, epithelial cell membrane antigen (ECMA), hypoxia inducible factor (HIF-1), carbonic anhydrase IX (CAIX), laminin, fibrin, fibronectin, fibroblast growth factor, transforming growth factors (TGF), fibroblast activation protein (FAP), tissue inhibitors of metalloproteinases (TIMPs), nitric oxide synthase (NOS), inducible and endothelial NOS, lysosomes in cells, macrophages, neutrophils, lymphocytes, hepatocyte growth factor (HGF), anti-neuropeptides, neutral endopeptidase (NEP), granulocyte-macrophage colony stimulating factor (GMCSF), neutrophil elastases, cathepsins, arginases, fibroblasts, endothelial cells and keratinocytes, keratinocyte growth factor (KGF), macrophage inflammatory protein-2 (MIP-2), macrophage inflammatory protein-2 (MIP-2), and macrophage chemoattractant protein-1 (MCP-1), polymorphonuclear neutrophils (PMN), macrophages, myofibroblasts, interleukin-1 (IL-1), tumour necrosis factor (TNF), nitric oxide (NO), c-myc, beta-catenin, endothelial progenitor cells (EPCs), matrix metalloproteinases (MMPs) and MMP inhibitors.
9 . The method of claim 1 , wherein illuminating the target with excitation light having at least one wavelength or wavelength band causing the one or more pre-selected biomarkers to fluoresce comprises illuminating the target with excitation light having a wavelength between 400 nm and 450 nm.
10 . The method of claim 1 , wherein detecting fluorescence emissions of the one or more pre-selected biomarkers with the image detector of the handheld imaging device includes filtering emissions responsive to illumination of the target with one or more filters configured to permit optical signals having wavelengths corresponding to bacterial fluorescence to pass through the one or more filters to the image detector of the handheld device.
11 . The method of claim 10 , wherein the wavelengths corresponding to bacterial fluorescence include one or more of wavelengths of 490 nm to 550 nm and wavelengths above 600 nm.
12 . The method of claim 1 , wherein identifying the one or more of the presence, the location, and the quantity of the biological contamination on or in the target comprises displaying the detected fluorescence emissions on a display of the handheld imaging device.
13 . The method of claim 12 , further comprising displaying an image of one or more of colonies of bacteria and biological fluids detected on or in the target.
14 . The method as claimed in claim 13 , wherein displaying the image of the one or more of colonies of bacteria and biological fluids detected on or in the target includes displaying the image on a display of the handheld device.
15 . The method of claim 13 , wherein displaying the image of the one or more of colonies of bacteria and biological fluids detected on the target comprises displaying an image of a location or a biodistribution of the one or more of the colonies of bacteria and biological fluids on the target.
16 . The method of claim 1 , wherein the target is contained in one or more of a public setting, a health care setting, a forensic setting, a clinical setting, a sterile setting, a laboratory setting, a surgical setting, an environmental setting, a hospital, a clinic, a nursing home, a home for aging adults, a surgical suite, and a bathroom.
17 . The method of claim 1 , wherein the target is selected from the group consisting of a surgical field, a skin target, an environmental target, a forensic target, a laboratory target, a surgical instrumentation target, a biological target, and a non-biological target.
18 . The method of claim 1 , further comprising, when one or more of the presence, the location, and the quantity of the biological contamination er on or in the target is identified, removing the identified biological contamination from the target.
19 . The method of claim 18 , further comprising, subsequent to removing the identified biological contamination from the target:
illuminating the target with excitation light emitted by the excitation light source of the handheld imaging device;
detecting fluorescence emissions of the one or more pre-selected biomarkers with the image detector of the handheld imaging device;
illuminating the target with white light emitted by the white light source of the handheld imaging device;
detecting reflection from the target in response to illumination of the target with the white light; and
displaying a new representation of the target based on the detected fluorescence emissions and detected reflection.
20 . The method of claim 1 , further comprising storing, archiving, cataloguing, or retrieving from storage the displayed representation of the target.
21 . The method of claim 1 , further comprising comparing previously calculated fluorescence intensities of the target stored on a memory of the handheld imaging device and an external database to newly calculated fluorescence intensities of the target.
22 . The method of claim 1 , further comprising:
cleaning the target, and
subsequent to the cleaning of the target, identifying and displaying one or more of a presence, a location, and a quantity of remaining biological contamination on the cleaned target based on the detected fluorescence emissions of the one or more pre-selected biomarkers.
23 . The method of claim 21 , further comprising identifying a change in one or more of the presence, the location, and the quantity of one or more of the colonies of bacteria and the biological fluid contaminating the target based at least in part on a comparison of the previously calculated fluorescence intensities of the target and the newly calculated fluorescence intensities of the target.
24 . The method of claim 1 , wherein the representation of the target comprises fluorescence intensity data displayed in combination with reflectance data.