Wide-field Fourier ptychographic microscopy using laser illumination source

Fourier ptychographic (FP) microscopy is a coherent imaging method that can synthesize an image with a higher bandwidth using multiple low-bandwidth images captured at different spatial frequency regions. The method’s demand for multiple images drives the need for a brighter illumination scheme and a high-frame-rate camera for a faster acquisition. We report the use of a guided laser beam as an illumination source for an FP microscope. It uses a mirror array and a 2-dimensional scanning Galvo mirror system to provide a sample with plane-wave illuminations at diverse incidence angles. The use of a laser presents speckles in the image capturing process due to reflections between glass surfaces in the system. They appear as slowly varying background fluctuations in the final reconstructed image. We are able to mitigate these artifacts by including a phase image obtained by differential phase contrast (DPC) deconvolution in the FP algorithm. We use a 1-Watt laser configured to provide a collimated beam with 150 mW of power and beam diameter of 1 cm to allow for the total capturing time of 0.96 seconds for 96 raw FPM input images in our system, with the camera sensor’s frame rate being the bottleneck for speed. We demonstrate a factor of 4 resolution improvement using a 0.1 NA objective lens over the full camera field-of-view of 2.7 mm by 1.5 mm

Extracellular Localisation of the C-Terminus of DDX4 Confirmed by Immunocytochemistry and Fluorescence-Activated Cell Sorting

Putative oogonial stem cells (OSCs) have been isolated by fluorescence-activated cell sorting (FACS) from adult human ovarian tissue using an antibody against DEAD-box helicase 4 (DDX4). DDX4 has been reported to be germ cell specific within the gonads and localised intracellularly. White et al. (2012) hypothesised that the C-terminus of DDX4 is localised on the surface of putative OSCs but is internalised during the process of oogenesis. This hypothesis is controversial since it is assumed that RNA helicases function intracellularly with no extracellular expression. To determine whether the C-terminus of DDX4 could be expressed on the cell surface, we generated a novel expression construct to express full-length DDX4 as a DsRed2 fusion protein with unique C- and N-terminal epitope tags. DDX4 and the C-terminal myc tag were detected at the cell surface by immunocytochemistry and FACS of non-permeabilised human embryonic kidney HEK 293T cells transfected with the DDX4 construct. DDX4 mRNA expression was detected in the DDX4-positive sorted cells by RT-PCR. This study clearly demonstrates that the C-terminus of DDX4 can be expressed on the cell surface despite its lack of a conventional membrane-targeting or secretory sequence. These results validate the use of antibody-based FACS to isolate DDX4-positive putative OSCs

Unshielded Two-Wire Circuit Systems under Weak Unbalance for High-Intensity Radiated Field Radiated Susceptibility by Double Bulk Current Injection

In unshielded two-wire circuits of weapon systems, the equipment at both ends of the cable needs to be tested at the same time. The theoretical model of substituting double bulk current injection (DBCI) for high-intensity radiated field (HIRF) electromagnetic radiation is established, which aims to calculate the relationship between the injection excitation voltage source and the radiation field intensity. Additionally, the conditions required for linear extrapolation is clarified. Subsequently, the test method of using DBCI for the HIRF radiated susceptibility (RS) is proposed. Next, the verification of the CST studio and the pass-through load test are performed. All results shows that the test method can avoid the influence of the impedance parameters of the two equipment at both ends, which can be applied to nonlinear terminals for the HIRF RS test

Feasibility and Error Analysis of Using Fiber Optic Temperature Measurement Device to Evaluate the Electromagnetic Safety of Hot Bridge Wire EEDs

Most studies assessing the safety of hot bridge wire EEDs employ temperature sensors that directly use the measurements of the temperature measurement device without analyzing the accuracy of the temperature measurement. This study establishes the response function of the exposed bridge and exposed bridge temperature rise system of hot bridge wire EEDs through the Rosenthal’s temperature rise equation and Laplace transformation as well as experimental tests, and through the response function, the response law and numerical characteristics of the two are compared and analyzed under four typical excitations. Under steady current injection and continuous-wave radiation, both exposed bridge and exposed bridge temperature measurement systems can reach thermal equilibrium, and the equilibrium temperature of both are the same. However, under pulse excitation, the temperature rise measurement value is significantly different from the actual value due to the large difference in response time of the exposed bridge (1 ms) and the exposed bridge temperature measurement device (0.82 s). Studies have shown that under steady current injection and continuous-wave radiation, temperature rise measurements can be directly applied to the safety assessment of hot bridge wire EEDs, while under pulsed conditions, temperature rise measurements cannot be directly applied

Transcriptomic survey of key reproductive and metabolic tissues in mouse models of polycystic ovary syndrome

Transcriptomic mapping of key reproductive and metabolic tissues and oocytes in mouse models of polycystic ovary syndrome reveals that the hypothalamus is susceptible to fetal programming while adipose and ovary are more affected upon postnatal androgen exposure

Single-cell analysis of human ovarian cortex identifies distinct cell populations but no oogonial stem cells

The human ovary orchestrates sex hormone production and undergoes monthly structural changes to release mature oocytes. The outer lining of the ovary (cortex) has a key role in defining fertility in women as it harbors the ovarian reserve. It has been postulated that putative oogonial stem cells exist in the ovarian cortex and that these can be captured by DDX4 antibody isolation. Here, we report single-cell transcriptomes and cell surface antigen profiles of over 24,000 cells from high quality ovarian cortex samples from 21 patients. Our data identify transcriptional profiles of six main cell types; oocytes, granulosa cells, immune cells, endothelial cells, perivascular cells, and stromal cells. Cells captured by DDX4 antibody are perivascular cells, not oogonial stem cells. Our data do not support the existence of germline stem cells in adult human ovaries, thereby reinforcing the dogma of a limited ovarian reserve. The outer lining or cortex of the human ovary determines fertility and has been postulated to contain oogonial stem cells. Here, the authors generate a single-cell map of the adult human ovarian cortex and show that DDX4 labels perivascular cells but no oogonial stem cells.Peer reviewe

Prenatal androgen exposure causes a sexually dimorphic transgenerational increase in offspring susceptibility to anxiety disorders

If and how obesity and elevated androgens in women with polycystic ovary syndrome (PCOS) affect their offspring’s psychiatric health is unclear. Using data from Swedish population health registers, we showed that daughters of mothers with PCOS have a 78% increased risk of being diagnosed with anxiety disorders. We next generated a PCOS-like mouse (F0) model induced by androgen exposure during late gestation, with or without diet-induced maternal obesity, and showed that the first generation (F1) female offspring develop anxiety-like behavior, which is transgenerationally transmitted through the female germline into the third generation of female offspring (F3) in the androgenized lineage. In contrast, following the male germline, F3 male offspring (mF3) displayed anxiety-like behavior in the androgenized and the obese lineages. Using a targeted approach to search for molecular targets within the amygdala, we identified five differentially expressed genes involved in anxiety-like behavior in F3 females in the androgenized lineage and eight genes in the obese lineage. In mF3 male offspring, three genes were dysregulated in the obese lineage but none in the androgenized lineage. Finally, we performed in vitro fertilization (IVF) using a PCOS mouse model of continuous androgen exposure. We showed that the IVF generated F1 and F2 offspring in the female germline did not develop anxiety-like behavior, while the F2 male offspring (mF2) in the male germline did. Our findings provide evidence that elevated maternal androgens in PCOS and maternal obesity may underlie the risk of a transgenerational transmission of anxiety disorders in children of women with PCOS.CC BY 4.0Correspondence: Elisabet Stener-Victorin ([email protected])</p

Excess of ovarian nerve growth factor impairs embryonic development and causes reproductive and metabolic dysfunction in adult female mice

Nerve growth factor (NGF) is critical for the development and maintenance of the peripheral sympathetic neurons. NGF is also involved in the ovarian sympathetic innervation and in the development and maintenance of folliculogenesis. Women with the endocrine disorder, polycystic ovary syndrome (PCOS), have an increased sympathetic nerve activity and increased ovarian NGF levels. The role of ovarian NGF excess in the PCOS pathophysiology and in the PCOS-related features is unclear. Here, using transgenic mice overexpressesing NGF in the ovarian theca cells (17NF mice), we assessed the female embryonic development, and the reproductive and metabolic profile in adult females. Ovarian NGF excess caused growth restriction in the female fetuses, and a delayed gonocyte and primary oocyte maturation. In adulthood, the 17NF mice displayed irregular estrous cycles and altered ovarian expression of steroidogenic and epigenetic markers. They also exhibited an increased sympathetic output with increased circulating dopamine, and metabolic dysfunction reflected by aberrant adipose tissue morphology and function, impaired glucose metabolism, decreased energy expenditure, and hepatic steatosis. These findings indicate that ovarian NGF excess leads to adverse fetal development and to reproductive and metabolic complications in adulthood, mirroring common features of PCOS. This work provides evidence that NGF excess may be implicated in the PCOS pathophysiology. CC BY-NC-ND 4.0</p