scholarly journals Optogenetic pacing in Drosophila melanogaster

2015 ◽  
Vol 1 (9) ◽  
pp. e1500639 ◽  
Author(s):  
Aneesh Alex ◽  
Airong Li ◽  
Rudolph E. Tanzi ◽  
Chao Zhou
Keyword(s):  
Drosophila Melanogaster ◽  
Electrical Stimulation ◽  
High Speed ◽  
Developmental Stages ◽  
Imaging System ◽  
Model Organism ◽  
Heart Rhythm ◽  
Cardiac Pacing ◽  
Microscopy Imaging ◽  
Cardiac Tissues

Electrical stimulation is currently the gold standard for cardiac pacing. However, it is invasive and nonspecific for cardiac tissues. We recently developed a noninvasive cardiac pacing technique using optogenetic tools, which are widely used in neuroscience. Optogenetic pacing of the heart provides high spatial and temporal precisions, is specific for cardiac tissues, avoids artifacts associated with electrical stimulation, and therefore promises to be a powerful tool in basic cardiac research. We demonstrated optogenetic control of heart rhythm in a well-established model organism, Drosophila melanogaster. We developed transgenic flies expressing a light-gated cation channel, channelrhodopsin-2 (ChR2), specifically in their hearts and demonstrated successful optogenetic pacing of ChR2-expressing Drosophila at different developmental stages, including the larva, pupa, and adult stages. A high-speed and ultrahigh-resolution optical coherence microscopy imaging system that is capable of providing images at a rate of 130 frames/s with axial and transverse resolutions of 1.5 and 3.9 μm, respectively, was used to noninvasively monitor Drosophila cardiac function and its response to pacing stimulation. The development of a noninvasive integrated optical pacing and imaging system provides a novel platform for performing research studies in developmental cardiology.

Fly-on-a-Chip: Microfluidics for Drosophila melanogaster Studies

2019 ◽  
Vol 11 (12) ◽  
pp. 425-443 ◽  
Author(s):  
Alireza Zabihihesari ◽  
Arthur J Hilliker ◽  
Pouya Rezai
Keyword(s):  
Drosophila Melanogaster ◽  
Developmental Stages ◽  
Model Organism ◽  
Fruit Fly ◽  
In Vitro Assays ◽  
Behavioral Studies ◽  
And Behavior ◽  
Manual Manipulation

Abstract The fruit fly or Drosophila melanogaster has been used as a promising model organism in genetics, developmental and behavioral studies as well as in the fields of neuroscience, pharmacology, and toxicology. Not only all the developmental stages of Drosophila, including embryonic, larval, and adulthood stages, have been used in experimental in vivo biology, but also the organs, tissues, and cells extracted from this model have found applications in in vitro assays. However, the manual manipulation, cellular investigation and behavioral phenotyping techniques utilized in conventional Drosophila-based in vivo and in vitro assays are mostly time-consuming, labor-intensive, and low in throughput. Moreover, stimulation of the organism with external biological, chemical, or physical signals requires precision in signal delivery, while quantification of neural and behavioral phenotypes necessitates optical and physical accessibility to Drosophila. Recently, microfluidic and lab-on-a-chip devices have emerged as powerful tools to overcome these challenges. This review paper demonstrates the role of microfluidic technology in Drosophila studies with a focus on both in vivo and in vitro investigations. The reviewed microfluidic devices are categorized based on their applications to various stages of Drosophila development. We have emphasized technologies that were utilized for tissue- and behavior-based investigations. Furthermore, the challenges and future directions in Drosophila-on-a-chip research, and its integration with other advanced technologies, will be discussed.

scholarly journals Non-invasive red-light optogenetic control of Drosophila cardiac function

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Jing Men ◽  
Airong Li ◽  
Jason Jerwick ◽  
Zilong Li ◽  
Rudolph E. Tanzi ◽  
...  
Keyword(s):  
Blue Light ◽  
Developmental Stages ◽  
Genetic Model ◽  
Imaging System ◽  
Red Light ◽  
Heart Rhythm ◽  
Light Stimulation ◽  
Excitation Light ◽  
Non Invasive ◽  
Recovery Dynamics

AbstractDrosophila is a powerful genetic model system for cardiovascular studies. Recently, optogenetic pacing tools have been developed to control Drosophila heart rhythm noninvasively with blue light, which has a limited penetration depth. Here we developed both a red-light sensitive opsin expressing Drosophila system and an integrated red-light stimulation and optical coherence microscopy (OCM) imaging system. We demonstrated noninvasive control of Drosophila cardiac rhythms using a single light source, including simulated tachycardia in ReaChR-expressing flies and bradycardia and cardiac arrest in halorhodopsin (NpHR)-expressing flies at multiple developmental stages. By using red excitation light, we were able to pace flies at higher efficiency and with lower power than with equivalent blue light excitation systems. The recovery dynamics after red-light stimulation of NpHR flies were observed and quantified. The combination of red-light stimulation, OCM imaging, and transgenic Drosophila systems provides a promising and easily manipulated research platform for noninvasive cardiac optogenetic studies.

scholarly journals The Phenotypic Effect of Cardamom oil on the Developmental Stages of Drosophila Melanogaster

2021 ◽  
pp. 225-231
Author(s):  
Dr. Y. D. Akhare ◽  
H. A. Patharikar
Keyword(s):  
Drosophila Melanogaster ◽  
Developmental Stages ◽  
Culture Media ◽  
Model Organism ◽  
Fruit Fly ◽  
Life Stages ◽  
Human Genetic Disease ◽  
Phenotypic Effect ◽  
Animal Development ◽  
Food And Drink

The fruit fly Drosophila melanogaster has been extensively studied as a model organism for genetic investigation. It also has many characteristics which make it an ideal organism for the study of animal development and behaviour, neurobiology and human genetic disease and condition. Drosophila melanogaster share several basic biological and chemical neurological and physiological similarities with mammals. In the present study, we noted the phenotypic effect of cardamom oil on the different stages of Drosophila melanogaster. The fruit flies were grown on 10-gram culture media supplemented with different concentration of cardamom oil (0.5µl, 1 µl, 2.5 µl). Further, the size and growth of different life stages of Drosophila melanogaster were observed and total protein estimated from it.The increase in the size and protein concentration in different life stages of controlled Drosophila melanogaster were recorded. Cardamom is a highly valued herbal spice used in tropical and subtropical Asia. cardamom is used as a flavouring and cooking spices in both food and drink and as a medicine.

scholarly journals Programmable RNA Targeting using CasRx in Flies

2020 ◽  
Author(s):  
Anna Buchman ◽  
Dan J. Brogan ◽  
Ruichen Sun ◽  
Ting Yang ◽  
Patrick Hsu ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Engineering ◽  
Developmental Stages ◽  
Comprehensive Evaluation ◽  
Model Organism ◽  
Endogenous Expression ◽  
Current State ◽  
Rna Targeting ◽  
Transcript Cleavage ◽  
Early Developmental

AbstractCRISPR-Cas genome editing technologies have revolutionized the fields of functional genetics and genome engineering, but with the recent discovery and optimization of RNA-targeting Cas ribonucleases, we may soon see a similar revolution in the study of RNA function and transcriptome engineering. However, to date, successful proof-of-principle for Cas ribonuclease RNA targeting in eukaryotic systems has been limited. Only recently has successful modification of RNA expression by a Cas ribonuclease been demonstrated in animal embryos. This previous work, however, did not evaluate endogenous expression of Cas ribonucleases and only focused on Cas ribonuclease function in early developmental stages. A more comprehensive evaluation of this technology is needed to assess its potential impact in the field. Here we report on our efforts to develop a programmable platform for RNA-targeting using a Cas ribonuclease, CasRx, in the model organism Drosophila melanogaster. By genetically encoding CasRx in flies, we demonstrate moderate transcript targeting of known phenotypic genes in addition to unexpected toxicity and lethality. We also report on the off-target effects following on-target transcript cleavage by CasRx. Taken together, our results present the current state and limitations of a genetically encoded programmable RNA-targeting Cas system in Drosophila melanogaster, paving the way for future optimization of the system.

scholarly journals Non-invasive red-light optogenetic control of Drosophila cardiac function

2020 ◽  
Author(s):  
Jing Men ◽  
Airong Li ◽  
Jason Jerwick ◽  
Zilong Li ◽  
Rudolph E. Tanzi ◽  
...  
Keyword(s):  
Blue Light ◽  
Developmental Stages ◽  
Genetic Model ◽  
Imaging System ◽  
Red Light ◽  
Heart Rhythm ◽  
Light Stimulation ◽  
Excitation Light ◽  
Non Invasive ◽  
Recovery Dynamics

ABSTRACTDrosophila is a powerful genetic model system for cardiovascular studies. Recently, optogenetic pacing tools have been developed to control Drosophila heart rhythm noninvasively with blue light, which has a limited penetration depth. Here we developed both a red-light sensitive opsin expressing Drosophila system and an integrated red-light stimulation and optical coherence microscopy (OCM) imaging system. We demonstrated noninvasive control of Drosophila cardiac rhythms, including simulated tachycardia in ReaChR-expressing flies and bradycardia and cardiac arrest in halorhodopsin (NpHR)-expressing flies at multiple developmental stages. By using red excitation light, we were able to pace flies at higher efficiency and with lower power than with equivalent blue light excitation systems. The recovery dynamics after red-light stimulation of NpHR flies were observed and quantified. The combination of red-light stimulation, OCM imaging, and transgenic Drosophila systems provides a promising and easily manipulated research platform for noninvasive cardiac optogenetic studies.

Spectral Microscopy Imaging System for High-Resolution and High-Speed Imaging of Fuel Sprays

2020 ◽  
Vol 142 (9) ◽  
Author(s):  
Ken Maassen ◽  
Farzad Poursadegh ◽  
Caroline Genzale
Keyword(s):  
High Resolution ◽  
High Speed ◽  
High Pressures ◽  
High Efficiency ◽  
Imaging System ◽  
Direct Injection ◽  
Resolving Power ◽  
Microscopy Imaging ◽  
Fuel Sprays ◽  
Primary Breakup

Abstract Modern high-efficiency engines utilize direct injection for charge preparation at extremely high pressures. At these conditions, the scales of atomization become challenging to measure, as primary breakup occurs on the micrometer and nanosecond scales. As such, fuel sprays at these conditions have proven difficult to study via direct imaging. While high-speed cameras now exist that can shutter at tens to hundreds of nanoseconds, and long-range microscopes can be coupled to these cameras to provide high-resolution images, the resolving power of these systems is typically limited by pixel size and field of view (FOV). The large pixel sizes make the realization of the diffraction-limited optical resolution quite challenging. On the other hand, limited data throughput under high repetition rate operation limits the FOV due to reduced sensor area. Therefore, a novel measurement technique is critical to study fuel spray formation at engine-relevant conditions. In this work, we demonstrate a new high-resolution imaging technique, spectral microscopy, which aims to realize diffraction-limited imaging at effective framerates sufficient for capturing primary breakup in engine-relevant sprays. A spectral microscopy system utilizing a consumer-grade DSLR allows for significantly wider FOV with improved resolving power compared to high-speed cameras. Temporal shuttering is accomplished via separate and independently triggered back illumination sources, with wavelengths selected to overlap with the detection bands of the camera sensor's RGB filter array. The RGB detection channels act as filters to capture independently timed red, green, and blue light pulses, enabling the capture of a three consecutive images at effective framerates exceeding 20 × 106 fps. To optimize system performance, a backlit illumination system is designed to maximize light throughput, a multilens setup is created, and an image-processing algorithm is demonstrated that formulates a three-frame image from the camera sensor. The system capabilities are then demonstrated by imaging engine relevant diesel sprays. The spectral microscopy system detailed in this paper allows for micron-scale feature recognition at framerates exceeding 20 × 106 fps, thus expanding the capability for experimental research on primary breakup in fuel sprays for modern direct-injection engines.

Smart Scanning Ion-Conductance Microscopy Imaging Technique Using Horizontal Fast Scanning Method

2018 ◽  
Vol 24 (3) ◽  
pp. 264-276 ◽  
Author(s):  
Jian Zhuang ◽  
Zhiwu Wang ◽  
Zeqing Li ◽  
Pengbo Liang ◽  
Mugubo Vincent
Keyword(s):  
High Speed ◽  
Imaging System ◽  
Scanning Speed ◽  
Transverse Motion ◽  
Acquisition Time ◽  
Microscopy Imaging ◽  
Ion Conductance ◽  
Scanning Method ◽  
Scanning Ion Conductance Microscopy

AbstractTo solve extended acquisition time issues inherent in the conventional hopping-scanning mode of scanning ion-conductance microscopy (SICM), a new transverse-fast scanning mode (TFSM) is proposed. Because the transverse motion in SICM is not the detection direction and therefore presents no collision problem, it has the ability to move at high speed. In TSFM, the SICM probe gradually descends in the vertical/detection direction and rapidly scans in the transverse/nondetection direction. Further, the highest point that decides the hopping height of each scanning line can be quickly obtained. In conventional hopping mode, however, the hopping height is artificially set without a priori knowledge and is typically very large. Consequently, TFSM greatly improves the scanning speed of the SICM imaging system by effectively reducing the hopping height of each pixel. This study verifies the feasibility of this novel scanning method via theoretical analysis and experimental study, and compares the speed and quality of the scanning images obtained in the TFSM with that of the conventional hopping mode. The experimental results indicate that the TFSM method has a faster scanning speed than other SICM scanning methods while maintaining the quality of the images. Therefore, TFSM provides the possibility to quickly obtain high-resolution three-dimensional topographical images of extremely complex samples.

Dual-stage piezo nanopositioner for high-speed ion conductance microscopy imaging

2020 ◽  
Vol 28 (10) ◽  
pp. 2203-2214
Author(s):  
Jian ZHUANG ◽  
◽  
Zhi-wu WANG ◽  
Xiao-bo LIAO ◽  
Keyword(s):  
High Speed ◽  
Microscopy Imaging ◽  
Ion Conductance ◽  
Dual Stage

scholarly journals FlyBase: updates to the Drosophila melanogaster knowledge base

2020 ◽  
Vol 49 (D1) ◽  
pp. D899-D907 ◽  
Author(s):  
Aoife Larkin ◽  
Steven J Marygold ◽  
Giulia Antonazzo ◽  
Helen Attrill ◽  
Gilberto dos Santos ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Knowledge Base ◽  
Fast Track ◽  
Model Organism ◽  
Disease Models ◽  
Online Database ◽  
Experimental Tool

Abstract FlyBase (flybase.org) is an essential online database for researchers using Drosophila melanogaster as a model organism, facilitating access to a diverse array of information that includes genetic, molecular, genomic and reagent resources. Here, we describe the introduction of several new features at FlyBase, including Pathway Reports, paralog information, disease models based on orthology, customizable tables within reports and overview displays (‘ribbons’) of expression and disease data. We also describe a variety of recent important updates, including incorporation of a developmental proteome, upgrades to the GAL4 search tab, additional Experimental Tool Reports, migration to JBrowse for genome browsing and improvements to batch queries/downloads and the Fast-Track Your Paper tool.

Sign in / Sign up

Export Citation Format

Share Document