Handheld Digital Microscope Research Articles

Assessment of Nailfold Capillaroscopy Findings in Pregnant Women having Preeclampsia: A Research Protocol

Introduction: Pregnancy causes significant vascular and cutaneous changes in a woman’s body. Preeclampsia is one of the life-threatening conditions associated with high blood pressure and proteinuria after the 20th week of pregnancy. It can cause placental hypoperfusion and placental insufficiency, leading to foetal defects or, in severe cases, foetal loss, which can be prevented by early diagnosis and treatment. There are many ways to observe and study the microcirculation non invasively the safest and most accessible method is nailfold capillaroscopy. Nail-fold Video Capillaroscopy (NVC) is a handheld digital microscope used to observe the microcirculation at the nailfold of the hands. Thus, comparing the capillary status with healthy subjects will help establish an early diagnosis and manage cases to prevent further complications in the mother and newborn. Need of the Study: The pathophysiology states that preeclampsia is related to systemic endothelial dysfunction, causing the release of Tumour Necrosis Factor (TNF) alpha, a proinflammatory mediator, leading to hypoxia of the placenta and the loss of endothelial cell-lined capillaries. Thus, there is a need to find if there is an established correlation in association of decreased Capillary Density (CD) in individuals with preeclampsia. Although evidence supporting a link between decreased CD and preeclampsia is limited, timely assessment and prevention are crucial. Nailfold capillaroscopy, an accessible and non invasive imaging method, holds promising calibre for understanding microvascular changes in preeclampsia. This could facilitate post-conception monitoring and early interventions to identify and address circulatory issues in affected women with preeclampsia. Aim: To study and compare the nailfold capillary changes in women diagnosed with preeclampsia and women having normal and uneventful pregnancies. Materials and Methods: A comparative cross-sectional study will be done in the Outpatient Department (OPD) and Inpatient Department (IPD) of Obstetrics and Gynaecology and the Outpatient Department of Dermatology at AVBRH Sawangi Meghe, Wardha, Maharashtra, India, from March 2023 to March 2025. A total of 150 pregnant women will be considered in the study, with 75 women being normal pregnant women (controls) and 75 pregnant women diagnosed with preeclampsia (cases). In order to confirm the gestational age of the patients, fundal height and abdominal circumference measurements will be taken. Demographic data, Basal Metabolic Index (BMI), weight gain during pregnancy, laboratory reports including Complete Blood Count (CBC), coagulation profile (Prothrombin Time, International Normalised Ratio, Activated Partial Thromboplastin Time), liver function tests, kidney function tests, urine examinations, and serial ultrasonography reports available for the patients will be noted. Pre-examination blood pressure will be recorded and compared with the baseline to confirm the status of preeclampsia after obtaining written consent from the patients. Nailfold capillaroscopy will be performed using a (Dinolite -AF4115ZT) dermoscope equipped with a 200X magnification, high-resolution lens in a non polarised light setting with oil immersion as a fluid coupling. The observations will be used to interpret the results of the study. Results will be calculated for finding differences between the two comparative groups of cases and controls at a p-value of ≤0.05.

A Marble Bust Newly Discovered by the Trapani Lombardo Family of Reggio Calabria (Southern Italy)

This research concerns a stylistic and archaeometric study of an ancient marble female bust recently discovered by the Trapani Lombardo family of Reggio Calabria (Southern Italy) and delivered to the Superintendency of Archaeology, Fine Arts, and Landscape for the metropolitan city of Reggio Calabria and Vibo Valentia (SABAP). Based on the first technical, stylistic, and iconographic observations made by the competent bodies, it is a half-length portrait bust from the Roman era, which precisely had the function of faithfully reproducing the physiognomy of the depicted subject. The research aimed to establish the authenticity of the artwork and the origin of the raw material, providing indications about the textural and compositional features and of the alteration products as well as identifying traces of any previous restoration interventions. For these purposes, after a preliminary assessment of the state of conservation of the bust using visual inspections supported by a handheld digital microscope, different analytical techniques, including polarized optical microscopy (OM), scanning electron microscopy coupled with energy dispersive spectrometry (SEM-EDX), carbon and oxygen stable isotope ratio determinations (δ13C and δ18O), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FT-IR), were used. The results highlighted the originality of the artifact, thus remarking on the importance of the precious archaeological find to be included in the cataloging of tangible assets in the panorama of Italian cultural heritage.

A Novel Approach to Tally Aculops cannabicola (Acari: Eriophyidae) for Field and Laboratory Studies.

The hemp russet mite, Aculops cannabicola (Farkas) is a key pest of hemp (Cannabis sativa L.). Given its microscopic size, estimating the size of its populations can be a major limitation to control this pest. Here, we describe a method to count A. cannabicola using photographs taken with a handheld digital microscope. We compared the consistency and strength of the relationship between photographic- and microscope-based counts of mites from infested plants. Among the three sections of the leaflet, the maximum consistency levels were observed in the proximal (50%) and middle photographs (50%). However, it reached from 64% to 100% when the three sections were considered. Photographic and a microscope-based counts were positively correlated (>0.7). A single photograph can be a good predictor of the total mites per leaflet, however, using two pictures (proximal and middle sections) will increase the consistency of the abundance of A. cannabicola per leaflet. A minimum of 22 leaflets per sampling event can support a strong correlation between the microscope and photographic counts. Our method requires low budget and training and takes short time (0.4 to 1.3 min per leaflet) to count mites per sample. Additionally, photographs can be stored on a smartphone, computer, or tablet, allowing users to share, store and process the photos. This method simplifies counts of A. cannabicola on hemp for research purposes and provides a practical tool for growers to assess mite populations for management decisions. In addition, it may be useful for monitoring eriophyid mites on cultivated plants.

Paper-based sol-gel thin films immobilized cytochrome P450 for enzyme activity measurement

Cytochrome P450 (CYP450), and in particular CYP3A4, is the most abundantly expressed CYP450 isozyme implicated in many drug-drug and medicinal plant-drug interactions. Therefore, incorporation of CYP3A4 enzyme screening at an early stage of drug discovery is preferable in order to avoid enzymatic interactions. Here we present for the first time a paper-based CYP3A4 immobilized sol-gel-derived a platform using resorufin benzyl ether as a fluorogenic enzyme substrate used to investigate enzyme activity. The fluorescence intensity of the product can be simply quantified by using a handheld digital microscope and an image analysis software. The limit of quantitation was 0.35 μM with good precision (RSDs < 4.1%). Furthermore, the assay of CYP3A4 activity on the developed paper-based device provided comparable results with those obtained from conventional well-plates (p > 0.05), while offering simplicity and lower cost. Kinetic parameters of the immobilized CYP3A4 in sol-gel coated paper were calculated from the Lineweaver-Burk plot, including Michaelis constant (Km) and maximum velocity (Vmax), which were 2.71 ± 0.35 μM and 0.43 ± 0.05 μM/min, respectively. Moreover, a functional test of these devices was conducted by assessments of known CYP3A4 inhibitors (i.e. ketoconazole, itraconazole) and inducers (i.e. phenytoin, carbamazepine). To further demonstrate the broad range of uses, the devices were utilized to assay plant extracts i.e. Areca catechu seeds, Camellia sinensis leaves, Eclipta prostrata aerial part, providing results in good agreement with previous studies. Furthermore, the sol-gel immobilized enzyme stored at 4 °C can increase storage stability, offering the activity of 86.3 ± 0.4% after 3-weeks storage, equivalent to the activity of the free enzyme solution after 1-week storage. The developed paper-based devices offer versatility, portability and low-cost.

In Situ Diagnosis of Scabies Using a Handheld Digital Microscope in Resource-Poor Settings-A Proof-of-Principle Study in the Amazon Lowland of Colombia.

Scabies is a neglected tropical disease associated with important morbidity. The disease occurs worldwide and is particularly common in resource-poor communities in the Global South. A validated technique for the diagnosis of scabies in resource-poor settings does not exist. The objective of the study was to determine the practicability and accuracy of handheld digital microscopy in three indigenous communities in the Amazon lowland of Colombia, where scabies is the most common parasitic skin disease. One-hundred-and-eleven children and adults from three indigenous communities with a presumptive diagnosis of scabies were examined clinically by using a handheld digital microscope placed directly on the skin. The microscopical identification of a mite was verified by an “experienced mother”, a woman who had acquired the skills to diagnose scabies as part of traditional Amerindian medicine. The “experienced mother” removed the parasite with a fine needle and placed it on a flat surface in order to enable its direct examination with the digital microscope. Using digital microscopy, scabies was diagnosed in 24 out of 111 participants and confirmed by the extraction of a Sarcoptes mites from the acarine eminence. A characteristic tunnel (burrow) with or without mite could be clearly identified irrespective of the degree of pigmentation of the skin. Besides, digital microscopy revealed pathological characteristics of scabies hitherto unknown and impossible to be seen in dermoscopy, such as partially or totally obliterated tunnels, tunnels with multiple entry or exit points, circumscribed hyperpigmentation around obliterated tunnels and mites secluded in a nodule. This proof-of-principle study demonstrated the accurate diagnosis of scabies by handheld digital microscopy in patients with pigmented skin and the feasibility of this technique in resource-poor settings.

An Improved In Vivo Methodology to Visualise Tumour Induced Changes in Vasculature Using the Chick Chorionic Allantoic Membrane Assay.

Decreasing the vascularity of a tumour has proven to be an effective strategy to suppress tumour growth and metastasis. Anti-angiogenic therapies have revolutionized the treatment of advanced-stage cancers, however there is still demand for further improvement. This necessitates new experimental models that will allow researchers to reliably study aspects of angiogenesis. The aim of this study was to demonstrate an in vivo technique in which the highly vascular and accessible chorioallantoic membrane (CAM) of the chick embryo is used to study tumour-induced changes in the macro and microvessels. Two cancer cell lines (human melanoma (C8161) and human prostate cancer (PC3)) were selected as model cells. Human dermal fibroblasts were used as a control. One million cells were labelled with green fluorescent protein and implanted on the CAM of the chick embryo at embryonic development day (EDD) 7 and angiogenesis was evaluated at EDDs 10, 12 and 14. A fluorescently-tagged lectin (lens culinaris agglutinin (LCA)) was injected intravenously into the chick embryo to label endothelial cells. The LCA is known to label the luminal surface of endothelial cells, or dextrans, in the CAM vasculature. Macrovessels were imaged by a hand-held digital microscope and images were processed for quantification. Microvessels were evaluated by confocal microscopy. Tumour invasion was assessed by histological and optical sectioning. Tumour cells (C8161 and PC3) produced quantifiable increases in the total area covered by blood vessels, compared to fibroblasts when assessed by digital microscopy. Tumour invasion could be demonstrated by both histological and optical sectioning. The most significant changes in tumour vasculature observed were in the microvascular structures adjacent to the tumour cells, which showed an increase in the endothelial cell coverage. Additionally, tumour intravasation and tumour thrombus formation could be detected in the areas adjacent to tumour cells. The fragility of tumour blood vessels could be demonstrated when tumour cells seeded on a synthetic scaffold were grown on CAM. We report on a modification to a well-studied CAM in vivo assay, which can be effectively used to study tumour induced changes in macro and microvasculature.

Microscopic Surface Measurement Using Phase Shifting Method

Accurate information of microscopic topography is very important for efficacy assessment of a surface texture of skin health. Due to the limitations of the direct visual assessment of skin microscopic topography, an optical dermastocopy is very common to be used as skin imaging device to magnify skin topography based on a white light reflection. The limitation of this method is its poor spatial resolution to quantify skin topography. In this work, microscopic skin imaging based on phase shifting method is configured using a DLP pico-projector with LED illumination and a handheld digital microscope. As illuminator for the digital microscope, the DLP projector is programmed to generate patterned light on skin surface. Image processing is required in providing accurate information of surface topography. The first step, a wrapped phase shifting must be extracted from acquired intensity images. The second step is obtaining unwrapped phase image, which is a critical process because it must be recovered from wrapped phase shifting that containednoise. Finally, phase offset due to multiples of 2π during phase unwrapping must be removed. Early experiments on simple object are carried out to test the level of distortion of fringe in several variations of contrast and also to test the performance of the system on several frequency variations. The test results indicate the depth proportion obtained from absolute phase image has the same trend as the proportion of direct measurement. Implementation on the skin surface profile performed on three test areas: the back of the hand and knuckle creases. Based on quantitative and qualitative analysis,our proposed scheme of skin imaging based on phase shifting is promising for surface profile measurement and imaging of the skin.

Paper-based 3D microfluidic device for multiple bioassays

This study investigates a new paper-based 3D microfluidic analytical device for analyzing multiple biological fluids. A wax-printed and -impregnated device was operated using tip-pinch manipulation of the thumb and index fingers and applied the chemical reaction of a preloaded colorimetric indicator and biological solutions. Chemical sensing of protein and glucose concentrations was quantitatively analyzed by changes in the color intensity of the image taken from three image readout devices including scanner (Epson Perfection V700), microscope (USB-embedded handheld digital microscope), and smartphone (LG Optimus Vu). Paper-based 3D microfluidic analytic device with three image analyzers successfully quantified 1.5–75μM protein concentrations and 0–900mg/dL glucose concentrations. Paper-based 3D microfluidic device combined with the smartphone showed the performance in protein bioassay (1.5–75μM) and glucose bioassay (0–50mM) including clinically relevant ranges comparable to other devices. An origami-driven paper-based 3D microfluidic analytic is a useful platform with great potential for application in point-of-care diagnostics.

A preliminary study of the contribution of telemicroscopy to the diagnosis and management of skin tumours in teledermatology

Dermatology images can be acquired using a miniature handheld digital microscope at magnifications of 10- to 30-fold. We investigated the contribution of telemicroscopy to the reliability and accuracy of teledermatology for skin tumours. In a retrospective study of the reliability of diagnosis and management plans, two dermatologists compared teledermatology with face-to-face assessment. We also measured the accuracy of diagnosis by comparing teledermatology with histopathological examination. Two dermatologists evaluated the clinical images and information for 120 randomly sorted cases and recorded a diagnosis and a management plan for each case. Two months later, the same dermatologists re-evaluated the cases and suggested a new diagnosis and management plan for each case. The reliability of teledermatology diagnosis was not significantly different from face-to-face examination; it was significantly increased with the addition of microscopic images (P < 0.001). The reliability of teledermatology management plans was not significantly different from face-to-face examination; it was significantly increased with the addition of the microscopic images (P < 0.001). The accuracy of teledermatology was significantly increased with microscopic images (P = 0.05). Telemicroscopy appears to be a useful adjunct to teledermatology with a digital camera. Future studies with different magnifications should be performed to detect the optimum magnification for telemicroscopy.

Handheld digital microscope with the latest LCD TV monitor

1. Dae-young Kim, MD, PhD 2. Hyung Suk Kim, MD (td8144{at}yahoo.co.kr) 1. Seoul, Korea In the graft dissection process of hair transplantation, optical microscopes are widely used as a method to lower follicle transection rates. However, long working hours at the microscope stand and limited

The Pixel Paradox and Transition-Metal Spectroscopy: One of Many Uses of the Handheld Digital Microscope in Chemical Demonstrations

A handheld digital microscope (HDM) interfaced to a computer with a presentation projector is used to project an out-of-focus yellow patch on the screen, then the patch is brought into focus to show that, paradoxically, there are red and green but no yellow pixels. Chromaticity diagrams are used to discuss this observation and spectroscopic examples of additive mixing. Types of HDMs and other uses of these devices are described.