High Precision Ping-pong Auto-zeroed Lock-in Fluorescence Photometry Sensor.

This paper presents a high-precision CMOS fluorescence photometry sensor using a novel lock-in amplification scheme based on switched-biasing and ping-pong auto-zeroing techniques. The CMOS sensor includes two photodiodes and a lock-in amplifier (LIA) operating at 1 kHz. The LIA comprises a differential low-noise amplifier using a novel switched-biasing ping-pong auto-zeroed scheme, an automatic phase aligner, a programmable gain amplifier, a band-pass filter, a mixer, and an output low-pass filter. The design is fabricated in 0.18-µm CMOS process, and the measurement shows that the LIA can retrieve noisy input signals with a dynamic reserve of 42 dB, while consuming only 0.7 mW from a 1.8 V supply voltage. The measured results show that the LIA can detect a wide range of incident light power from 8 nW to 24 µW. The proposed design is encapsulated in a 3D-printed housing allowing for real-time in vitro biomarker detection. This ambulatory platform uses an LED and a fiber optic to convey the excitation light to the sample and retrieve the fluorescence signal. Experiments with a beads solution diluted in PBS demonstrate that the sensor has a sensitivity of 1:100 k. Experimental results obtained in vitro with NIH3T3 mouse cells tagged with membrane dye show the ability of the prototype to detect different densities of cell culture. The portable prototype, which includes optical filters and a small 30 mm × 36 mm × 30 mm printed circuit board enclosed inside the 3D-printed housing, consumes 36.7 mW and weighs 120 g.

Understanding the nervous system: lessons from Frontiers in Neurophotonics.

The Frontiers in Neurophotonics Symposium is a biennial event that brings together neurobiologists and physicists/engineers who share interest in the development of leading-edge photonics-based approaches to understand and manipulate the nervous system, from its individual molecular components to complex networks in the intact brain. In this Community paper, we highlight several topics that have been featured at the symposium that took place in October 2022 in Québec City, Canada.

A Phosphoinositide-Binding Protein Acts in the Trafficking Pathway of Hemoglobin in the Malaria Parasite Plasmodium falciparum.

Phosphoinositide lipids play key roles in a variety of processes in eukaryotic cells, but our understanding of their functions in the malaria parasite Plasmodium falciparum is still very much limited. To gain a deeper comprehension of the roles of phosphoinositides in this important pathogen, we attempted gene inactivation for 24 putative effectors of phosphoinositide metabolism. Our results reveal that 79% of the candidates are refractory to genetic deletion and are therefore potentially essential for parasite growth. Inactivation of the gene coding for a Plasmodium-specific putative phosphoinositide-binding protein, which we named PfPX1, results in a severe growth defect. We show that PfPX1 likely binds phosphatidylinositol-3-phosphate and that it localizes to the membrane of the digestive vacuole of the parasite and to vesicles filled with host cell cytosol and labeled with endocytic markers. Critically, we provide evidence that it is important in the trafficking pathway of hemoglobin from the host erythrocyte to the digestive vacuole. Finally, inactivation of PfPX1 renders parasites resistant to artemisinin, the frontline antimalarial drug. Globally, the minimal redundancy in the putative phosphoinositide proteins uncovered in our work supports that targeting this pathway has potential for antimalarial drug development. Moreover, our identification of a phosphoinositide-binding protein critical for the trafficking of hemoglobin provides key insight into this essential process. IMPORTANCE Malaria represents an enormous burden for a significant proportion of humanity, and the lack of vaccines and problems with drug resistance to all antimalarials demonstrate the need to develop new therapeutics. Inhibitors of phosphoinositide metabolism are currently being developed as antimalarials but our understanding of this biological pathway is incomplete. The malaria parasite lives inside human red blood cells where it imports hemoglobin to cover some of its nutritional needs. In this work, we have identified a phosphoinositide-binding protein that is important for the transport of hemoglobin in the parasite. Inactivation of this protein decreases the ability of the parasite to proliferate. Our results have therefore identified a potential new target for antimalarial development.

A pan-apicomplexan phosphoinositide-binding protein acts in malarial microneme exocytosis.

Invasion of human red blood cells by the malaria parasite Plasmodium falciparum is an essential step in the development of the disease. Consequently, the molecular players involved in host cell invasion represent important targets for inhibitor design and vaccine development. The process of merozoite invasion is a succession of steps underlined by the sequential secretion of the organelles of the apical complex. However, little is known with regard to how their contents are exocytosed. Here, we identify a phosphoinositide-binding protein conserved in apicomplexan parasites and show that it is important for the attachment and subsequent invasion of the erythrocyte by the merozoite. Critically, removing the protein from its site of action by knock sideways preferentially prevents the secretion of certain types of micronemes. Our results therefore provide evidence for a role of phosphoinositide lipids in the malaria invasion process and provide further insight into the secretion of microneme organelle populations, which is potentially applicable to diverse apicomplexan parasites.