This PDF file contains the front matter associated with SPIE Proceedings Volume 8951, including the Title Page, Copyright Information, Table of Contents, and the Conference Committee listing.

The developed measuring system is based on a low-coherence source and a self-mixing (or internal) detection. The proposed optical layout exploits the reflection from the internal wall of the duct as reference arm, thus reducing system complexity, cost, size and increasing its robustness to movements of the measurand. Moreover, the usage of a low-coherence source allows reducing the problems related to the poor definition of the volume under test (sensing region or measurement volume) typical of “coherent” self-mixing systems. Although preliminary analysis have been performed by simply investigating the frequencies relative to the maximum in the Doppler spectrum, the obtained results demonstrates that by increasing scatterers concentration of +300%, the system sensitivity increases of about only +20%.

Photoplethysmography is a technique widely used in monitoring perfusion and blood oxygen saturation based on the amplitude of the pulsatile signal at one or multiple wavelengths. However, the pulsatile signal carries in its waveform a substantial amount of information about the mechanical properties of the tissue and vasculature under investigation that is still yet to be utilized to its full potential. In this work, we present the feasibility of pulse wave analysis for the application of monitoring hepatic implants and diagnosing graft complications. In particular, we demonstrate the utility of computing the slope of the pulse during the diastole phase to assess compliance changes in tissue. This hypothesis was tested in a series of in vitro experiments using a polydimethylsiloxane based phantom mimicking the optical and mechanical properties of the portal vein. The emptying time decreased from 148.1 ms for phantoms with compliance of 12 KPa to 97.5 ms for phantoms with compliance of 61 KPa. These compliance levels mimic those seen for normal and fibrotic hepatic tissue respectively.

In this paper we demonstrate a novel application of correlation mapping optical coherence tomography (cm-OCT) for volumetric nailfold capillaroscopy (NFC). NFC is a widely used non-invasive diagnostic method to analyze capillary morphology and microvascular abnormalities of nailfold area for a range of disease conditions. However, the conventional NFC is incapable of providing volumetric imaging, when volumetric quantitative microangiopathic parameters such as plexus morphology, capillary density, and morphologic anomalies of the end row loops most critical. cm-OCT is a recently developed well established coherence domain magnitude based angiographic modality, which takes advantage of the time-varying speckle effect, which is normally dominant in the vicinity of vascular regions compared to static tissue region. It utilizes the correlation coefficient as a direct measurement of decorrelation between two adjacent B-frames to enhance the visibility of depth-resolved microcirculation.

Millions of people worldwide are affected by diabetes. While glucose sensing technology has come a long way over the past several decades, the current commercially available techniques are still invasive, often leading to poor patient compliance. To minimize invasiveness, focus has been placed on optical techniques to ascertain blood glucose concentrations. Optical polarimetry has shown promise and progress as a viable technique for glucose sensing. Recent developments in polarimetric glucose sensing have been focused on overcoming time varying corneal birefringence due to motion artifacts. Beyond corneal birefringence, the next hurdle toward making this approach viable is the ability to couple polarized light across the eye’s anterior chamber. The eye is ideally suited to couple light to the retina. The index mismatch between the air and cornea is partially responsible for the beam bending toward the retina and, while good for vision, it complicates our ability to couple light across the anterior chamber without using an index matching device when performing polarimetric glucose monitoring. In this report, we have designed and modeled a non-index matched coupling scheme constructed with commercially available optics. The optical ray tracing model was performed using CODE V to verify the feasibility of a reflective based non-index matched coupling scheme with respect to index of refraction and anatomical restraints. The ray tracing model was developed for a dual-wavelength system and the effect of refraction and reflection at each optical interface within the setup was evaluated. The modeling results indicate a reflective based optical coupling design could be added to existing polarimetric glucose systems thus removing the need for placing an index matched eye-coupling mechanism over the eye prior to data collection.

In many publications, infrared spectroscopy excelled in multi-analyte assays of biofluids. Based on such technology, laboratory and point-of-care applications can be envisaged and most needed devices are for blood glucose measurements. Implementing strict glycemic control can reduce the risk of serious complications in both diabetic and critically ill patients. For this purpose, many different blood glucose monitoring techniques and insulin infusion strategies have been tested towards the realization of an artificial pancreas under closed loop control. However, for patient portable instrumentation current FTIR-spectrometers are still too bulky, which need replacement by devices that allow further miniaturization. Recently developed external cavity quantum cascade lasers (EC-QCL) are tuneable over about 200 cm^-1, which however is still narrow, compared to the range accessible with FTIR-devices. In this work, we applied bandwidth constraints to previous FTIR-studies on blood plasma and dialysates of biofluids. For the clinically most important blood glucose no impairment was found using one laser only, provided that specific interferents were missing. Other analytes of interest, such as lactate and urea, indicated the need of broader tuneability over about 500 cm^-1 with a second or third laser for a simultaneous glucose assay.

Measurement of blood analytes provides crucial information about a patient’s health. Some such analytes, such as glucose in the case of diabetes, require long-term or near-continuous monitoring for proper disease management. However, current monitoring techniques are far from ideal: multiple-per-day finger stick tests are inconvenient and painful for the patient; implantable sensors have short functional life spans (i.e., 3-7 days). Due to analyte transporters on red blood cell (RBC) membranes that equilibrate intracellular and extracellular analyte levels, RBCs serve as an attractive alternative for encapsulating analyte sensors. Once reintroduced to the blood stream, the functionalized RBCs may continue to live for the remainder of their life span (120 days for humans). They are biodegradable and biocompatible, thereby eliminating the immune system response common for many implanted devices. The proposed sensing system utilizes the ability of the RBCs to swell in response to a decrease in the osmolarity of the extracellular solution. Just before lysis, they develop small pores on the scale of tens of nanometers. While at low temperature, analyte-sensitive dyes in the extracellular solution diffuse into the perforated RBCs and become entrapped upon restoration of temperature and osmolarity. Since the fluorescent signal from the entrapped dye reports on changes in the analyte level of the extracellular solution via the RBC transporters, interactions between the RBCs and the dye are critical to the efficacy of this technique. In this work, we study the use of a near infrared pH sensitive dye encapsulated within RBCs and assess the ability to measure dye fluorescence in vivo.

Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA⁺) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a Förster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.

We present the digital inline holographic microscopy for blood cell counting. The microscope unit contains a pinhole chip for illumination, a micro-fluidic chip, and a CMOS-chip for detection. The design is adapted and tested for blood cell diagnostics. A single hologram delivers a stack of images for all z-positions in the micro-fluidic channel and is the basic for counting of flowing cells. Images of flowing red blood cells are presented providing clinical diagnostics.

We propose an optical cavity based biosensor with chained differential detection. A three laser diode sensing mechanism provides multiplexing capability and is used to enhance the responsivity and improve fabrication tolerance using a chained differential detection approach. The differential calculation enhances the sensitivity through (1) increased responsivity compared to the actual optical power changes of the individual laser diodes and (2) noise reduction by canceling out some uncontrollable variations along the path of light since all wavelengths of light used for the differential calculation propagate through the same path. However, the responsivity dies off quickly due to even small variations from the designed cavity width. To correct for this and to improve fabrication tolerance, we introduce another wavelength and employ a chaining approach. In this presentation, we will present simulation results of an optical cavity based biosensor with chained differential detection and progresses toward experimental demonstrations.

We introduce a fluorescent imaging method that is capable of detecting fluorescent micro-particles over an ultra-wide field of view of 19 cm × 28 cm using a modified flatbed scanner. We added a custom-designed absorbing emission filter, a computer controlled two dimensional LED array, and modified the driver of the scanner to maximize the sensitivity, exposure time, and gain for fluorescent detection of micro-objects. This high-throughput fluorescent imaging device used in conjunction with a microfluidic sample holder enables rapid screening of fluorescent micro-objects inside more than 2.2mL of optically dense media (i.e., whole blood) within 5 minutes. The device is sensitive enough to detect fluorescently labeled cells, and generates images that have an effective pixel count of 2.2 Giga-pixels.

Due to its effectiveness and broad coverage, Ciprofloxacin is the fifth most prescribed antibiotic in the US. As current methods of infection diagnosis and antibiotic sensitivity testing (i.e. an antibiogram) are very time consuming, physicians prescribe ciprofloxacin before obtaining antibiogram results. In order to avoid increasing resistance to the antibiotic, a method was developed to provide both a rapid diagnosis and the sensitivity to the antibiotic. Using Surface Enhanced Raman Spectroscopy, an antibiogram was obtained after exposing the bacteria to Ciprofloxacin for just two hours. Spectral analysis revealed clear separation between sensitive and resistant bacteria and could also offer some inside into the mechanisms of resistance.

In this paper, I demonstrate a low cost light weight microscope device that is compatible with any smartphone camera. The device amplifies the imaging resolution of a smartphone camera by three orders of magnitude from millimeters to sub-micrometers, while costing approximately USD$ 2.

In this paper, we study the photoconductivity of a polymer-based TiOPc (Titanium Oxide Phthalocyanine) thin-film for the development of a multi-opto-piezoelectric-valve-array. Using a polymer-based TiOPc thin film to serve as the electrode and a structural layer of a piezoelectric polymer, P(VDF-TrFE) poly[(vinylidenefluoride-co-trifluoroethylene], an optical control valve-array could be developed for manipulating multiple microdroplets for the application of digital microfluidic. In this ongoing project, the dependency of the light intensity, thickness, and composition of spin-coated polymer-based TiOPc thin-film was studied. The experimental finding suggested that a 14 to 55 times resistivity change could be achieved by controlling the film thickness to be between 0.9 μm and 1.5 μm with TiOPc concentration of 20% and 30% w/w compositions.

Erythrocyte zinc protoporphyrin-IX (ZnPP) and protoporphyrin-IX (PPIX) accumulate in a variety of disorders that restrict or disrupt the biosynthesis of heme, including iron deficiency and various porphyrias. We describe a reagent-free spectroscopic method based on dual-wavelength excitation that can measure simultaneously both ZnPP and PPIX fluorescence from unwashed whole blood while virtually eliminating background fluorescence. We further aim to quantify ZnPP and PPIX non-invasively from the intact oral mucosa using dual-wavelength excitation to reduce the strong tissue background fluorescence while retaining the faint porphyrin fluorescence signal originating from erythrocytes. Fluorescence spectroscopic measurements were made on 35 diluted EDTA blood samples using a custom front-face fluorometer. The difference spectrum between fluorescence at 425 nm and 407 nm excitation effectively eliminated background autofluorescence while retaining the characteristic porphyrin peaks. These peaks were evaluated quantitatively and the results compared to a reference HPLC-kit method. A modified instrument using a single 1000 μm fiber for light delivery and detection was used to record fluorescence spectra from oral mucosa. For blood measurements, the ZnPP and PPIX fluorescence intensities from the difference spectra correlated well with the reference method (ZnPP: Spearman’s rho r_s = 0.943, p < 0.0001; PPIX: r_s = 0.959, p < 0.0001). In difference spectra from oral mucosa, background fluorescence was reduced significantly, while porphyrin signals remained observable. The dual-wavelength excitation method evaluates quantitatively the ZnPP/heme and PPIX/heme ratios from unwashed whole blood, simplifying clinical laboratory measurements. The difference technique reduces the background fluorescence from measurements on oral mucosa, allowing for future non-invasive quantitation of erythrocyte ZnPP and PPIX.

There is a critical unmet clinical need for a device that can monitor and predict the onset of shock: hemorrhagic shock or bleeding to death, septic shock or systemic infection, and cardiogenic shock or blood flow and tissue oxygenation impairment due to heart attack. Together these represent 141 M patients per year. We have developed a monitor for shock based on measuring blood flow in peripheral (skin) capillary beds using diffuse correlation spectroscopy, a form of dynamic light scattering, and have demonstrated proof-of-principle both in pigs and humans. Our results show that skin blood flow measurement, either alone or in conjunction with other hemodynamic properties such as heart rate variability, pulse pressure variability, and tissue oxygenation, can meet this unmet need in a small self-contained patch-like device in conjunction with a hand-held processing unit. In this paper we describe and discuss the experimental work and the multivariate statistical analysis performed to demonstrate proof-of-principle of the concept.

Cervical cancer is a prevalent disease in many developing countries. Colposcopy is the most common approach for screening cervical intraepithelial neoplasia (CIN). However, its clinical efficacy heavily relies on the examiner’s experience. Spectroscopy is a potentially effective method for noninvasive diagnosis of cervical neoplasia. In this paper, we introduce a hyperspectral imaging technique for noninvasive detection and quantitative analysis of cervical neoplasia. A hyperspectral camera is used to collect the reflectance images of the entire cervix under xenon lamp illumination, followed by standard colposcopy examination and cervical tissue biopsy at both normal and abnormal sites in different quadrants. The collected reflectance data are calibrated and the hyperspectral signals are extracted. Further spectral analysis and image processing works are carried out to classify tissue into different types based on the spectral characteristics at different stages of cervical intraepithelial neoplasia. The hyperspectral camera is also coupled with a lab microscope to acquire the hyperspectral transmittance images of the pathological slides. The in vivo and the in vitro imaging results are compared with clinical findings to assess the accuracy and efficacy of the method.

The esophageal cancer has a tendency to transfer to another part of the body and the surgical operation itself sometimes gives high risk in vital function because many delicate organs exist near the esophagus. So the esophageal cancer is a disease with a high mortality. So, in order to lead a higher survival rate five years after the cancer’s treatment, the investigation of the diagnosis methods or techniques of the cancer in an early stage and support the therapy are required. In this study, we performed the ex vivo experiments to obtain the Raman spectra from normal and early-stage tumor (stage-0) human esophageal sample by using Raman spectroscopy. The Raman spectra are collected by the homemade Raman spectrometer with the wavelength of 785 nm and Raman probe with 600-um-diameter. The principal component analysis (PCA) is performed after collection of spectra to recognize which materials changed in normal part and cancerous pert. After that, the linear discriminant analysis (LDA) is performed to predict the tissue type. The result of PCA indicates that the tumor tissue is associated with a decrease in tryptophan concentration. Furthermore, we can predict the tissue type with 80% accuracy by LDA which model is made by tryptophan bands.

The aim of the present study is to evaluate the capability of a miniaturized Raman endoscope (mRE) system to monitor the advancement of colorectal tumors in live model mice. The endoscope is narrow enough to observe the inside of the mouse colon under anesthesia. The mRE system allows to observe the tissues and to apply a miniaturized Raman probe for the measurement at any targeted point within the colon. Raman spectroscopy allows obtaining information about molecular composition without damaging the tissue (i.e., noninvasively). Continuous monitoring of the same tumor is carried out to study molecular alterations along with its advancement. The Raman spectra measured before and after the anticancer drug (5-FU) treatment indicated spectral changes in the tumor tissue. It suggests that the tumor is not cured but supposedly transformed to another tumor type after the treatment.

A novel multi-wavelength photoplethysmograph (PPG), previously utilized to quantify optically absorptive circulating gold nanoparticles, has demonstrated the potential to enhance therapeutic treatment predictability as pharmacokinetic metrics are provided throughout the intravenous delivery phase of quinine in real-time. This report demonstrates how the PPG could be used to assess the real-time bioavailability of other types of intravenously delivered optically-absorbing nanoparticles and drugs. The drug currently under investigation is anti-malarial quinine (absorption peak ~350 nm). We describe how the algorithm has been adapted to quantify the concentration of quinine in the pulsatile, circulating blood based on its extinction at three wavelengths (340, 660 and 940 nm). We show an example of the system collecting data representing the baseline, injection, and the clearance phases. An examination of the raw signal suggests that the system is well suited to sense the concentration of quinine in the therapeutic range (10mg/kg).

Live subcutaneous tumor grown in nude mouse is studied in situ with hyperspectral autofluorescence imaging and Raman spectroscopy. The purpose of the study is to develop methods for characterization of biochemical changing and of histological type of tumor without labeling. The results show that there are site depending variation in the fluorescence and Raman spectra. At the spot in which calcification is in process, Raman spectra showed a strong and specific band at 957 cm^-1 due to PO₄ species. The autofluosescence image can prove the histological changes based on the NADH and FAD which are major fluorophores in biological tissues. The hyperspectral image is analyzed with principal component analysis and the reconstructed images successfully depicts a different between necrotic and viable part within living subcutaneous tumor.

During the perioperative period, which includes the period before surgery and after surgery (postoperative), it is essential to measure diagnostic parameters such as: blood oxygen saturation; hemoglobin (Hb) concentration; and pulse rate. The Hb concentration in human blood is an important parameter to evaluate the physiological condition of an individual, as Hb is the oxygen carrying component of red blood cells. By determining the Hb concentration, it is possible, for example, to observe intraoperative or postoperative bleeding, and use this information as a trigger for autologous/ allogenic blood transfusions. In blood donation center it is also an essential parameter for the decision regarding the acceptance of the donor.

Laser speckle contrast imaging technique has been playing an important role in monitoring cutaneous microcirculation, but the strong scattering of skin restricts the imaging depth and contrast, and also makes it impossible to assess the cutaneous microcirculation response dynamically with high sensitivity. The tissue optical clearing is helpful for opening a visible window on mouse dorsal skin. In this work, the cutaneous microcirculation response to vasoactive noradrenaline is monitored with the laser speckle contrast imaging system before and after skin optical clearing. The results show that the optical clearing method can significantly enhance the contrast of laser speckle contrast imaging, and small blood vessels whose diameter less than 20μm can be distinguished with high resolution. The dynamic changes in cutaneous microvascular diameter and blood flow velocity caused by drug can be monitored sensitively. In contrast, it is difficult to detect the cutaneous microcirculation response that occurred in the blood vessels more than 100μm in the intact skin, and the signal-to-noise ratio is too low to monitor the dynamic changes caused by the same drug. Thus, skin optical clearing method can enhance the ability of laser speckle contrast imaging in accessing cutaneous microcirculation response, including the imaging contrast, resolution and sensitivity.

We report on a method for analysing multispectral images of skin in vivo for the measurement and visualisation of skin characteristics. Four different indices were used to characterise skin tissue oxygenation, erythema, total haemoglobin and melanin content. Index values were calculated pixel-wise and combined to create index maps to visualise skin properties. Quantitative measurement of tissue oxygenation saturation was possible by calibrating the oxygenation index using a commercial, calibrated oximeter. Index maps were tested by arterial occlusion of the index finger with multispectral images taken before, during and after occlusion in a pilot study with 10 healthy controls.

In daily-life environment, the quantitative measurement of biological substances, such as the blood glucose level in the human skin, is strongly required to realize the non-invasive healthcare apparatus. Fourier-spectroscopic-tomography of the little-finger-size with high time-resolution and with the strong robustness for mechanical vibrations is proposed. The proposed method is a kind of near-common-path interferometer with spatial phase-shift method. We install the transmission-type relative-inclined phase-shifter on the optical Fourier transform plane of the infinity corrected optical system. The phase shifter is constructed with the cuboid and wedge prisms to give the relative phase-shift spatially between each half-flux of the objective beams. The interferograms from each single-bright-point on an objective surface in a line are formed as fringe patterns on 2-dimensional imaging array devices. And because the proposed method is based on the imaging optics, only emitted rays from a focal plane can contribute forming of interferograms. Thus, the measurement plane can be limited onto the focal plane only. From the spectroscopic tomography, only at a localized vessel area in human skins, we can get the pinpointed near-infrared spectroscopic data. And we can expect the improvement of the determination precision, because a Fourier spectroscopic-character is acquired from multiple intensity data in accordance with amount of phase-shift. From the statistical point of view, the gradation of detector is improved with the square root of sample number, based on t-distribution. We constructed the statistical model to assure the determination accuracy, and demonstrated the feasibility of the glucose sensor using liquid cells.

We are aiming at the realization of the measurement technology for the biological-substance distributions, such as sebum, on entire faces at the daily-life environment. We proposed the imaging-type 2-dimensional Fourier spectroscopy [1] that is the palmtop-size portable measurement apparatus and has the strong robustness for mechanical vibrations. And the proposed method can measure the wide-field 2-dimensional middle-infrared spectroscopic-imaging of radiation lights emitted from human bodies itself without light sources. In the proposed method, we install the phase-shifter, that can give an arbitrary phase difference for the half-flux of objective beams, at the optical Fourier transform plane of the infinity corrected optical system. The near-common-path interferometer that is a phase-shift interferometer between objective beams can be realized. In this proposed method, the emitted rays from each single-bright-point on measurement surfaces can interfere with each other. Thus, even if the middle infrared-lights from human bodies are the spatially incoherent light, we can acquire the interferograms at each pixel on an imaging array-device in accordance with the amount of phase shift as the 2-dimensional image-intensity changes. We demonstrated the feasibility of the middle infrared spectroscopic imaging of whole human faces without active illuminations.

Here, we show that multiplexed Whispering Gallery Modes sensing can be achieved using two dye-doped microspheres positioned onto the tip of a Microstructured Optical Fiber. By operating the dye-doped microspheres below their lasing threshold, the individual resonances of each sphere overlap and therefore cannot be distinguished. However, when excited above their lasing threshold results in a de-convoluted spectrum where the resonances belonging to the individual spheres can be determined, enabling the detection of a specific interaction in one resonator and using the second as a dynamic reference, or monitoring different specific interactions with both spheres.

Optical polarimetry in the anterior chamber of the eye has emerged as a potential technique to non-invasively measure glucose levels for diabetes. Time varying corneal birefringence due to eye motion artifact confounds the optical signal ultimately limiting the polarimetric technique from accurately predicting glucose concentrations. In this study, a high speed dual-wavelength optical polarimetric approach was developed and in vitro phantom studies were performed with and without motion. The glucose concentrations ranged from 0-600 mg/dL at 100 mg/dL increments. The polarimeter produced glucose measurements with less than a 10 msec stabilization time and yielding standard errors of less than 10 mg/dL without motion and standard errors less than 26 mg/dL with motion. The results indicate a high speed dual-wavelength polarimetric approach has the potential to be used for non-invasive glucose measurements.

The ability of people with diabetes to both monitor and regulate blood sugar levels is limited by the conventional “finger-prick” test that provides intermittent, single point measurements. Toward the development of a continuous glucose monitoring (CGM) system, the lectin, Concanavalin A (ConA), has been utilized as a component in a Förster resonance energy transfer (FRET), competitive glucose binding assay. Recently, to avoid reversibility problems associated with ConA aggregation, a suitable competing ligand labeled with 8-aminopyrene-1,3,6-trisulfonic acid trisodium salt (APTS) has been engineered. However, its ability to function as part of a glucose sensing assay is compromised due to the negative charge (at physiological pH) of native ConA that gives rise to non-specific binding with other ConA groups as well as with electrostatically charged assay-delivery carriers. To minimize these undesirable interactions, we have conjugated ConA with monomethoxy-poly(ethylene glycol) (mPEG) (i.e. “PEGylation”). In this preliminary research, fluorescently-labeled ConA was successfully PEGylated with mPEG-Nhydroxylsuccinimide( succinimidyl carbonate) (mPEG-NHS(SC)). The FRET response of APTS-labeled competing ligand (donor) conveyed an increase in the fluorescence intensity with increasing glucose concentrations.

It is challenging to track the rapid changes in drug concentrations after intra-arterial (IA) administration to elucidate the pharmacokinetics of this method of drug delivery. Traditional pharmacokinetic parameters (such as protein binding) that are highly relevant to intravenous (IV) administration do not seem to apply to IA injections. Regional drug delivery is affected by the biomechanics of drug injection, resting blood flow, and local tissue extraction. In-vivo and ex-vivo, optical methods for spatial mapping of drug deposition can assist in visualizing drug distributions and aid in the screening of potential drugs and carrier candidates. We present a multimodal approach for the assessment of drug distribution in postmortem tissue specimens using diffuse reflectance spectroscopy, multispectral imaging, and confocal microscopy and demonstrate feasibility of distinguishing route of administration advantages of liposome-dye conjugate delivery. The results of this study suggest that insight on drug dynamics gained by this aggregated approach can be used to help screen and/or optimize potential drug candidates and drug delivery protocols.

For diabetics, continuous glucose monitoring and the resulting tighter control of glucose levels ameliorate serious complications from hypoglycemia and hyperglycemia. Diabetics measure their blood glucose levels multiple times a day by finger pricks, or use implantable monitoring devices. Still, glucose and other analytes in the blood fluctuate throughout the day and the current monitoring methods are invasive, immunogenic, and/or present biodegradation problems. Using carrier erythrocytes loaded with a fluorescent sensor, we seek to develop a biodegradable, efficient, and potentially cost effective method to continuously sense blood analytes. We aim to reintroduce sensor-loaded erythrocytes to the bloodstream and conserve the erythrocytes lifetime of 120 days in the circulatory system. Here, we compare the efficiency of two loading techniques: hypotonic dilution and electroporation. Hypotonic dilution employs hypotonic buffer to create transient pores in the erythrocyte membrane, allowing dye entrance and a hypertonic buffer to restore tonicity. Electroporation relies on controlled electrical pulses that results in reversible pores formation to allow cargo entrance, follow by incubation at 37°C to reseal. As part of the cellular characterization of loaded erythrocytes, we focus on cell size, shape, and hemoglobin content. Cell recovery, loading efficiency and cargo release measurements render optimal loading conditions. The detected fluorescent signal from sensor-loaded erythrocytes can be translated into a direct measurement of analyte levels in the blood stream. The development of a suitable protocol to engineer carrier erythrocytes has profound and lasting implications in the erythrosensor’s lifespan and sensing capabilities.