Smartphone microscopic method for imaging and quantification of microplastics in drinking water.

Analysis of microplastics in drinking water is often challenging due to smaller particle size and low particle count. In this study, we used a low cost and an easy to assemble smartphone microscopic system for imaging and quantitating microplastic particles as small as 20 µm. The system consisted of a spherical sapphire ball lens of 4 mm diameter attached to a smartphone camera as a major imaging component. It also involved pre-concentration of the sample using ZnCl2 solution. The spike recovery and limit of detection of the method in filtered distilled and deionized water samples (n = 9) were 55.6% ± 9.7% and 34 particles/L, respectively. Imaging performance of the microscopic system was similar to a commercial bright field microscopic system. The method was further implemented to examine microplastic particles in commercial bottled and jar water samples (n = 20). The particles count in bottled and jar water samples ranged from 0-91 particles/L to 0-130 particles/L, respectively. In both sample types, particles of diverse shape and size were observed. The particles collected from water samples were further confirmed by FTIR spectra (n = 36), which found 97% of the particles tested were made of plastic material. These findings suggested that the smartphone microscopic system can be implemented as a low-cost alternative for preliminary screening of microplastic in drinking water samples. RESEARCH HIGHLIGHTS: Ball lens based smartphone microscopic method was used for microplastic analysis. Particles of diverse shape and size were found in bottle and jar water samples.

Characterization and removal of microplastics in the Guheshwori Wastewater Treatment Plant, Nepal.

Contamination of river water systems by microplastic particles (MPPs) is one of the emerging global environmental concerns with potentially widespread ecological, socioeconomic, and health implications. A wastewater treatment plant (WWTP) processes and treats wastewater to remove pollutants and release safe water into the environment. There has been limited research on the characterization of microplastics and their removal in WWTP in South Asia. In this work, we report on the characterization of microplastics in wastewater and sludge samples (n = 300) from Guheshwori WWTP located on the bank of the Bagmati River in Kathmandu city, Nepal representing inlet, secondary aeration tank (SAT), outlet, and sludge from November 2021 to November 2022. On average, we detected 31.2 ± 17.3 MPPs/L, 11.2 ± 9.4 MPPs/L, 8.5 ± 5.6 MPPs/L, and 6.6 ± 4.8 MPPs/g in the samples collected from inlet, SAT, outlet, and sludge, respectively. Commonly found MPPs were in the form of fiber, fragments, foam, and pellets. Largely, MPPs were red, yellow, white, blue, and black. Among the 44 µm - 150 µm, 150 µm - 500 µm and 500 µm - 5 mm categories of size fractions, the most dominant fractions were 500 µm - 150 µm in inlet, SAT, and sludge, and 150 µm - 44 µm in the outlet sampling unit. The Guheshwori WWTP was able to remove 72.5 % of MPPs on average, that mostly occurred in the inlet. The effluent released into the river and the sludge still contained a significant number of MPPs.

Himalayan lichen biomass for green synthesis of silver nanocolloids: growth kinetics, effect of pH and metal sensing.

Lichen is one of the most abundant non-vascular biomasses; however, a systematic study on the application of biomass in nanomaterial synthesis is very limited. In this study, an aqueous lichen extract was obtained from Hypotrachyna cirrhata, one of the most abundant Himalayan lichen biomasses, using a simple cold percolation method. The effects of extract-to-silver nitrate mixing ratio, pH and waiting time on the growth and stability of nanoparticles were systematically explored. The rate constant for bio-reduction was found to be 5.3 × 10-3 min-1. Transmission electron microscopy showed a narrow particle size distribution with a mean particle size of 11.1 ± 3.6 nm (n = 200). The X-ray diffraction and selected area electron diffraction techniques confirmed the formation of cubic crystals. The synthesized colloidal solution showed excellent response to Hg2+ and Cu2+ ions in spiked water samples. The limit of detection and calibration sensitivity for Hg2+ and Cu2+ ions were found to be 1 and 5 mg l-1 and 2.9 × 10-3 and 1.6 × 10-3 units ppm-1, respectively. These findings suggested that spherical silver nanoparticles with a narrow particle size distribution can be synthesized on a laboratory scale using an aqueous H. cirrhata lichen extract, and the colloidal solution can be used for the detection of selected heavy metals in water samples.

Water quality and hydrochemical assessments of thermal springs, Gandaki Province, Nepal.

Thermal springs are the sites where the water temperature lies above ambient temperature. They are widely used for power generation, hot water spas, balneotherapy, agriculture, laundering, and aquaculture. In Nepal, many thermal springs are reported but scientific understanding on water quality and hydrogeochemistry of the springs is very limited. In this study, a total of 28 physico-chemical parameters were measured in water samples collected from 12 thermal springs from Gandaki Province, Nepal. Correlation matrix and multivariate statistical analysis such as principal component analysis (PCA) and cluster analysis were used to understand the water quality and hydrogeochemistry of the hot water springs. The pH, temperature, electrical conductivity, total dissolved solids, and turbidity in the hot water springs ranged from 7.3 to 8.8, 31.6-64.3 °C, 206-16270 µS/cm, 115-6637 mg/L, and 0.21-63.7 NTU; respectively. The dominance order of major anions and cations were: Cl- > HCO3- > SO42- > NO3- > F- and Na+ > Ca2+ > K+ > Mg2+; respectively. Comparison of the water quality parameters with the WHO and National Water Quality Standards suggested that the majority of the parameters were within the safe limit. Out of 9 heavy metals and trace elements analyzed Zn2+, Ni2+, Cr3+, Cd+2, Hg (total), and Pb2+ were found below the safe limit but Fe (total), As (total) and Cu (total) were found higher than the WHO safe limit in total of 3, 5 and 1 sampling sites; respectively. The water quality index (WQI), sodium absorption ratio (SAR), and magnesium hazard (MH) ratio in the hot water springs ranged from 40.9 to 573, 2.2-49.3, 7.1-70.8; respectively. The result of PCA analysis showed that four principal components are required to explain hydrogeochemistry. Cluster analysis suggested that the sampling sites can be grouped into three distinct clusters based on total dissolved solids. Interestingly, the classification of hydrochemical facies using a Piper diagram suggested that 7 out of 12 thermal springs have Na-Cl type water. Finally, a perspective on the suitability of the hot springs for hot water spas and balneotherapy and policy recommendation is provided.

The Encapsulation of Bioactive Plant Extracts into the Cellulose Microfiber Isolated from G. optiva Species for Biomedical Applications.

Agricultural waste-based cellulose fibers have gained significant interest for a myriad of applications. Grewia optiva (G. optiva), a plant species, has been widely used for feeding animals, and the small branches' bark is used for making rope. Herein, we have extracted cellulose fibers from the bark of G. optiva species via chemical treatments (including an alkaline treatment and bleaching). The gravimetric analysis revealed that the bark of G. Optiva contains cellulose (63.13%), hemicellulose (13.52%), lignin (15.13%), and wax (2.8%). Cellulose microfibre (CMF) has been synthesized from raw fibre via chemical treatment methods. The obtained cellulose fibers were crosslinked and employed as the matrix to encapsulate the bioactive plant extracts derived from the root of Catharanthus roseus (C. roseus). The microscopic images, XRD, FTIR, and antibacterial/antioxidant activity confirmed the encapsulation of natural extracts in the cellulose microfiber. The microscopic images revealed that the encapsulation of the natural extracts slightly increased the fiber's diameter. The XRD pattern showed that the extracted cellulose microfiber had an average crystalline size of 2.53 nm with a crystalline index of 30.4% compared to the crystalline size of 2.49 nm with a crystalline index of 27.99% for the plant extract incorporated membrane. The water uptake efficiency of the synthesized membrane increased up to 250%. The antimicrobial activity of the composite (the CMF-E membrane) was studied via the zone inhibition against gram-positive and gram-negative bacteria, and the result indicated high antibacterial activity. This work highlighted G. optiva-derived cellulose microfiber as an optimum substrate for antimicrobial scaffolds. In addition, this paper first reports the antimicrobial/antioxidant behavior of the composite membrane of the C. roseus extract blended in the G. optiva microfiber. This work revealed the potential applications of CMF-E membranes for wound healing scaffolds.

Material Properties of Traditional Handmade Paper Samples Fabricated from Cellulosic Fiber of Lokta Bushes.

Handmade papers (HPs) are fabricated from fibrous biomass of Lokta bushes and other plant species following traditional eco-friendly method in Nepal. Although HP fabricated from Lokta bushes is believed to be durable and resistant to bugs and molds, material properties of this paper are not reported in literature. In this study, we measured several material properties of 10 handmade Lokta paper samples collected from local enterprises and paper industries. The mean caliper, grammage, apparent density, equilibrium moisture content, Cobb 60, brightness, opacity, tensile strength, and tensile index values in the paper samples ranged from ∼90 to 700 µm, 50 to 150 g/m2, 0.2 to 0.4 g/cm3, 4 to 7%, 50 to 400 g/m2, 56 to 67%, 83 to 98%, 30 to 2900 N/m, and 1 to 27 Nm/g, respectively. These properties suggested that the HPs are lightweight papers with intermediate to high strength. The tensile strength was found to be significantly higher along the length direction (p < 0.05). Characteristic features of cellulose, hemicellulose, and lignin were observed in FTIR spectra. The crystalline and amorphous phases were also identified in X-ray diffraction (XRD) data. Electron microscopy images revealed a nicely cross-linked network of intact fibers having almost parallel arrangement of microfibrils. These features could provide strength and durability to the paper samples. Understanding the material properties of HPs down to the sub-microscopic level may help improve the paper quality and find novel applications in the future.

Excessive iodine in iodized household salt in Nepal.

Iodine is an essential trace element required for the regulation of physiological processes involving the thyroid gland. However, inadequate and excessive intake of iodine are responsible for health problems, such as iodine deficiency disorders, hypothyroidism, hyperthyroidism, thyroiditis, thyroid papillary cancer, and thyrotoxicosis. The Universal Salt Iodization (USI) program has become successful in providing supplemental iodine at the population level globally. Packaging quality, fortification level, and transportation and storage conditions of iodized salt determine the availability of iodine. Previous studies have reported severe health issues caused by excessive iodine intake after the implementation of the USI program. To understand the levels of iodine, we collected 2117 household salt samples from seven districts of Nepal and tested them for iodine content; among them, 98.1% were iodized. Overall median concentration of iodine was 53.9 ppm (range: 43.5-61.4 ppm). The majority (67.2%) of samples had iodine in the range of 45-75 ppm. Approximately 0.9% of samples had inadequate, 13.3% contained adequate, and 83.9% had excessive iodine than the World Health Organization-recommended value. Iodine content varied among the sampling districts and seasons, to some extent. Our study confirmed that iodized salt is widely used in Nepal and is excessively iodized. Excessive intake of iodine through iodized salt requires further attention by policy makers. The iodine level may need adjustment to address the health impact.

Handmade Paper as a Paper Analytical Device for Determining the Quality of an Antidiabetic Drug.

Paper analytical devices (PADs) are a class of low-cost, portable, and easy-to-use platform for several analytical tests in clinical diagnostics, environmental pollution monitoring, and food and drug safety screening. These devices are primarily made from cellulosic paper. Considering the importance of eco-friendly and local or distributed manufacturing of devices realized during the COVID-19 pandemic, we systematically studied the potential of handmade Nepali paper to be used in fabricating PADs in this work. We characterized five different handmade papers made from locally available plant fibers using an eco-friendly method and used them to fabricate PADs for determining the drug quality. The thickness, grammage, and apparent density of the paper samples ranged from 198.6 to 314.8 µm, 49.1 to 117.8 g/m2, and 0.23 to 0.43 g/cm3, respectively. The moisture content, water filtration, and wicking speed ranged from 5.8 to 7.1%, 35.7 to 156.7, and 0.062 to 0.124 mms-1, respectively. Furthermore, the water contact angle and porosity ranged from 76.6 to 112.1° and 79 to 83%, respectively. The best paper sample (P5) was chosen to fabricate PADs for the determination of metformin, an antidiabetic drug. The metformin assay on PADs followed a linear range from 0.0625 to 0.5 mg/mL. The assay had a limit of detection and limit of quantitation of 0.05 and 0.18 mg/mL, respectively. The average amount of metformin concentration in samples collected from local pharmacies (n = 20) was 465.6 ± 15.1 mg/tablet. When compared with the spectrophotometric method, PAD assay correctly predicted the concentration of 90% samples. The PAD assay on handmade paper may provide a low-cost and easy-to-use system for screening the quality of drugs and other point-of-need applications.

Assessing volatile organic compound level in selected workplaces of Kathmandu Valley.

Volatile organic compounds (VOCs) are one of the major contributors to poor indoor air quality. Due to advancements in sensor technologies, continuous if not regular monitoring total VOC (TVOC) and or some specific VOC in potential high risk workplaces is possible even in resource limited settings. In this study, we implemented a portable VOC sensor to measure concentration of TVOC and formaldehyde (HCHO) in six types of potential high risk workplaces (n = 56 sites) of Katmandu Valley. For comparison, concentration was also measured in immediate surroundings (n = 56) of all the sites. To get preliminary information on safety practices, a survey study was also conducted. The mean TVOC and HCHO concentration in the sites ranged from 1.5‒8 mg/m3 and <0.01-5.5 mg/m3, respectively. The indoor: outdoor TVOC and HCHO ratio (I/O) was found to be significantly higher (I/O > 1.5 and p < 0.05) in 34 (~61%) and 47 sites (∼84%), respectively. A strong positive correlation between HCHO and TVOC concentration was observed in furniture industry (R = 0.91) and metal workshops (R = 0.98). Interestingly, we found TVOC and HCHO concentration higher than WHO safe limit in ∼64% and ∼32% sites, respectively. A rough estimate of chronic daily intake (CDI) of formaldehyde showed that CDI is higher than WHO limit in four sites. These findings suggested that indoor air quality in the significant number of the workplaces is poor and possible measures should be taken to minimize the exposure.

Cellulose-based micro-fibrous materials imaged with a home-built smartphone microscope.

Micro-fibrous materials are one of the highly explored materials and form a major component of composite materials. In resource-limited settings, an affordable and easy to implement method that can characterize such material would be important. In this study, we report on a smartphone microscopic system capable of imaging a sample in transmission mode. As a proof of concept, we implemented the method to image handmade paper samples-cellulosic micro-fibrous material of different thickness. With 1 mm diameter ball lens, individual cellulose fibers, fiber web, and micro-porous regions were resolved in the samples. Imaging performance of the microscopic system was also compared with a commercial bright field microscope. For thin samples, we found the image quality comparable to commercial system. Also, the diameter of cellulose fiber measured from both methods was found to be similar. We also used the system to image surfaces of a three ply surgical facemask. Finally, we explored the application of the system in the study of chemical induced fiber damage. This study suggested that the smartphone microscope system can be an affordable alternative in imaging thin micro-fibrous material in resource limited setting.

Review of analytical performance of COVID-19 detection methods.

In the recent SARS-CoV-2 pandemic, public health experts have emphasized testing, tracking infected people, and tracing their contacts as an effective strategy to reduce the spread of the virus. Several diagnostic methods are reported for detecting the coronavirus in clinical, research, and public health laboratories. Some tests detect the infection directly by detecting the viral RNA and other tests detect the infection indirectly by detecting the host antibodies. A diagnostic test during the pandemic should help make an appropriate clinical decision in a short period of time. Recently reported diagnostic methods for SARS-CoV-2 have varying throughput, batching capacity, requirement of infrastructure setting, analytical performance, and turnaround times ranging from a few minutes to several hours. These factors should be considered while selecting a reliable and rapid diagnostic method to help make an appropriate decision and prompt public health interventions. This paper reviews recent SARS-CoV-2 diagnostic methods published in journals and reports released by regulatory agencies. We compared the analytical efficiency including limit of detection, sensitivity, specificity, and throughput. In addition, we also looked into ease of use, affordability, and availability of accessories. Finally, we discuss the limitations of the methods and provide our perspectives on priorities for future test development.

A smartphone microscopic method for simultaneous detection of (oo)cysts of Cryptosporidium and Giardia.

BACKGROUND: Food and water-borne illness caused by ingestion of (oo)cysts of Cryptosporidium and Giardia is one of the major health problems globally. Several methods are available to detect Giardia cyst and Cryptosporidium oocyst in food and water. Most of the available methods require a good laboratory facility and well-trained manpower and are therefore costly. There is a need of affordable and reliable method that can be easily implemented in resource limited settings. METHODOLOGY/PRINCIPLE FINDINGS: We developed a smartphone based microscopic assay method to screen (oo)cysts of Cryptosporidium and Giardia contamination of vegetable and water samples. The method consisting of a ball lens of 1 mm diameter, white LED as illumination source and Lugols's iodine staining provided magnification and contrast capable of distinguishing (oo)cysts of Cryptosporidium and Giardia. The analytical performance of the method was tested by spike recovery experiments. The spike recovery experiments performed on cabbage, carrot, cucumber, radish, tomatoes, and water resulted in 26.8±10.3, 40.1±8.5, 44.4±7.3, 47.6±11.3, 49.2 ±10.9, and 30.2±7.9% recovery for Cryptosporidium, respectively and 10.2±4.0, 14.1±7.3, 24.2±12.1, 23.2±13.7, 17.1±13.9, and 37.6±2.4% recovery for Giardia, respectively. The spike recovery results are comparable with data obtained using commercial brightfield and fluorescence microscope methods. Finally, we tested the smartphone microscope system for detecting (oo)cysts on 7 types of vegetable (n = 196) and river water (n = 18) samples. Forty-two percent vegetable and thirty-nine percent water samples were found to be contaminated with Cryptosporidium oocyst. Similarly, thirty-one percent vegetable and thirty-three percent water samples were contaminated with Giardia cyst. CONCLUSIONS: The newly developed smartphone microscopic method showed comparable performance to commercial microscopic methods. The new method can be a low-cost and easy to implement alternative method for simultaneous detection of (oo)cysts in vegetable and water samples in resource limited settings.

A smartphone microscopic method for rapid screening of cloth facemask fabrics during pandemics.

BACKGROUND: In pandemics, because of increased demand and subsequent shortage of commercial facemasks, people need to use cloth facemasks, although such masks are reported to provide reduced protection. These masks can be prepared in local levels from different fabric materials. In developing countries, cloth masks are preferable because of low cost and added advantages of reusability. The filtering performance of a cloth facemask depends on the facial fit and on the material properties of fabrics such as porosity, yarn spacing or packing, and pore size. In resource limited settings, an affordable and easy to implement method that can assess the surface properties of cloth facemask fabrics would be important. METHODS: In this work, we developed a smartphone microscopic method for rapid screening of fabric quality. We measured the field of view of the microscope and as a proof of concept, we implemented the method to examine surfaces of sixteen locally available cloth mask fabrics. RESULTS: Out of the 16 masks examined, we found very diverse yarn packing and pore morphology (pore size and shape) in the fabrics. The pore size ranged from ~80 to 720 µm; much larger than respiratory droplet and bio-aerosol. This observation partly explains why such cloth facemasks provide reduced protection to the user during pandemics. The performance of a cloth facemask partly depends on the material properties of fabric such as yarn packing, pore size, porosity. Therefore, the surface properties of fabrics obtained from the smartphone method can be used to get preliminary idea on the facemask quality. We believe that the method can be an affordable and rapid method for selection of better fabrics for cloth facemask during pandemics.

Characterization of airborne dust samples collected from core areas of Kathmandu Valley.

Kathmandu Valley is reported to be one of the highly polluted and populated cities in the world. Particulate matter is one of the major contributors of unhealthy air in Kathmandu. Although there are several reports on spatial and temporal variation of air quality of Kathmandu Valley, the morphological and mineralogical characteristics of particulate matter are very limited or none. In this study, we report on the mineralogical and morphological analysis of airborne particulate matter collected from densely populated core areas of Kathmandu Valley using spectroscopic and microscopic techniques. The Fourier Transform Infrared (FTIR) and X-ray Diffraction (XRD) data showed the presence of clay minerals, crystalline silicate mineral, carbonate minerals, and asbestiform mineral in the dust samples. The field emission scanning electron microscopic analysis confirmed the existence of particles having diverse morphology with some of the particles having aspect ratio as high as twenty; indicating the existence of asbestiform type minerals. Based on SEM-EDX data, we found that the relative distribution of elements to be different in different samples and C, O, Mg, Ca, and Si were the major elements in the dust samples. Interestingly, the XRD data analysis showed that in all the samples quartz mineral having high degree of crystallinity was present. The XRD measurement was also carried out in three different brands of cement samples. Few minerals present in dust samples were also identified in the cement samples. This observation could indicate that cement is one of the sources of minerals in the airborne particulate matter in the Kathmandu Valley.

Optical microscopic study of surface morphology and filtering efficiency of face masks.

BACKGROUND: Low-cost face masks made from different cloth materials are very common in developing countries. The cloth masks (CM) are usually double layered with stretchable ear loops. It is common practice to use such masks for months after multiple washing and drying cycles. If a CM is used for long time, the ear loops become stretched. The loop needs to be knotted to make the mask loop fit better on the face. It is not clear how washing and drying and stretching practices change the quality of a CM. The particulate matter (PM) filtering efficiency of a mask depends on multiple parameters, such as pore size, shape, clearance, and pore number density. It is important to understand the effect of these parameters on the filtering efficiency. METHODS: We characterized the surface of twenty different types of CMs using optical image analysis method. The filtering efficiency of selected cloth face masks was measured using the particle counting method. We also studied the effects of washing and drying and stretching on the quality of a mask. RESULTS: The pore size of masks ranged from 80 to 500 µm, which was much bigger than particular matter having diameter of 2.5 µm or less (PM2.5) and 10 µm or less (PM10) size. The PM10 filtering efficiency of four of the selected masks ranged from 63% to 84%. The poor filtering efficiency may have arisen from larger and open pores present in the masks. Interestingly, we found that efficiency dropped by 20% after the 4th washing and drying cycle. We observed a change in pore size and shape and a decrease in microfibers within the pores after washing. Stretching of CM surface also altered the pore size and potentially decreased the filtering efficiency. As compared to CMs, the less frequently used surgical/paper masks had complicated networks of fibers and much smaller pores in multiple layers in comparison to CMs, and therefore had better filtering efficiency. This study showed that the filtering efficiency of cloth face masks were relatively lower, and washing and drying practices deteriorated the efficiency. We believe that the findings of this study will be very helpful for increasing public awareness and help governmental agencies to make proper guidelines and policies for use of face mask.

Status of chemistry lab safety in Nepal.

Chemistry labs can become a dangerous environment for students as the lab exercises involve hazardous chemicals, glassware, and equipment. Approximately one hundred thousand students take chemistry laboratory classes annually in Nepal. We conducted a survey on chemical lab safety issues across Nepal. In this paper, we assess the safety policy and equipment, protocols and procedures followed, and waste disposal in chemistry teaching labs. Significant population of the respondents believed that there is no monitoring of the lab safety in their lab (p<0.001). Even though many labs do not allow food and beverages inside lab and have first aid kits, they lack some basic safety equipment. There is no institutional mechanism to dispose lab waste and chemical waste is disposed haphazardly. Majority of the respondents believed that the safety training should be a part of educational training (p = 0.001) and they would benefit from short course and/or workshop on lab safety (p<0.001).

Investigating axial diffusion in cylindrical pores using confocal single-particle fluorescence correlation spectroscopy.

We explored the feasibility of using confocal fluorescence correlation spectroscopy to study small nanoparticle diffusion in hundred-nanometer-sized cylindrical pores. By modeling single particle diffusion in tube-like confined three-dimensional space aligned parallel to the confocal optical axis, we showed that two diffusion dynamics can be observed in both original intensity traces and the autocorrelation functions (ACFs): the confined two-dimensional lateral diffusion and the unconfined one-dimensional (1D) axial diffusion. The separation of the axial and confined lateral diffusion dynamics provides an opportunity to study diffusions in different dimensions separately. We further experimentally studied 45 nm carboxylated polystyrene particles diffusing in 300 nm alumina pores. The experimental data showed consistency with the simulation. To extract the accurate axial diffusion coefficient, we found that a 1D diffusion model with a Lorentzian axial collection profile needs to be used to analyze the experimental ACFs. The diffusion of the 45 nm nanoparticles in polyethyleneglycol-passivated 300 nm pores slowed down by a factor of ∼2, which can be satisfactorily explained by hydrodynamic frictions.

Nanosecond Time-Resolution Study of Gold Nanorod Rotation at the Liquid-Solid Interface.

Early studies showed that the adsorption of nanorods may start from a special "anchored" state, in which the nanorods lose translational motion but retain rotational freedom. Insight into how the anchored nanorods rotate should provide additional dimensions for understanding particle-surface interactions. Based on conventional time-resolution studies, gold nanorods are thought to continuously rotate following initial interactions with negatively charged glass surfaces. However, this nanosecond time-resolution study reveals that the apparent continuous rotation actually consists of numerous fast, intermittent rotations or transitions between a small number of weakly immobilized states, with the particle resting in the immobilized states most of the time. The actual rotation from one immobilized state to the other happens on a 1 ms timescale, that is, approximately 50 times slower than in the bulk solution.

Continuous-Wave Stimulated Emission Depletion Microscope for Imaging Actin Cytoskeleton in Fixed and Live Cells.

Stimulated emission depletion (STED) microscopy provides a new opportunity to study fine sub-cellular structures and highly dynamic cellular processes, which are challenging to observe using conventional optical microscopy. Using actin as an example, we explored the feasibility of using a continuous wave (CW)-STED microscope to study the fine structure and dynamics in fixed and live cells. Actin plays an important role in cellular processes, whose functioning involves dynamic formation and reorganization of fine structures of actin filaments. Frequently used confocal fluorescence and STED microscopy dyes were employed to image fixed PC-12 cells (dyed with phalloidin- fluorescein isothiocyante) and live rat chondrosarcoma cells (RCS) transfected with actin-green fluorescent protein (GFP). Compared to conventional confocal fluorescence microscopy, CW-STED microscopy shows improved spatial resolution in both fixed and live cells. We were able to monitor cell morphology changes continuously; however, the number of repetitive analyses were limited primarily by the dyes used in these experiments and could be improved with the use of dyes less susceptible to photobleaching. In conclusion, CW-STED may disclose new information for biological systems with a proper characteristic length scale. The challenges of using CW-STED microscopy to study cell structures are discussed.

A dual wavelength-activatable gold nanorod complex for synergistic cancer treatment.

A multifunctional gold nanorod (AuNR) complex is described with potential utility for theranostic anticancer treatment. The AuNR was functionalized with cyclodextrin for encapsulation of doxorubicin, with folic acid for targeting, and with a photo-responsive dextran-azo compound for intracellular controlled drug release. The interaction of a AuNR complex with HeLa cells was facilitated via a folic acid targeting ligand as displayed in the dark-field images of cells. Enhanced anticancer efficacy was demonstrated through the synergistic combination of promoted drug release upon ultraviolet (UV) light irradiation and photothermal therapy upon infrared (IR) irradiation. This multifunctional AuNR-based system represents a novel theranostic strategy for spatiotemporal delivery of anticancer therapeutics.