Name: an adjustable high-definition imaging system for behavioral studies of drosophila adults
Uploaded: 2021-06-08T13:30:00
Description: Watch this Scientific Journal Video about An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults at JoVE.com

We thank Professor Li Xiangdong and photographer Mr. Cheng Jing for helpful discussions and suggestions. This work was supported by the Exploratory Project (20200101) of the Life Science Experimental Teaching Demonstration Center of China Agricultural University.

1. Construction of the observing/documenting system
NOTE: The materials needed to construct the fly behavior observing/documenting system are shown in Figure 1, and the completed system is shown in Figure 2. The protocol to construct the system and how to use it are described below.
<ol>
	<li>Make a fly behavior observation chambe...

<ol>
	<li>Chyb, S., Gompel, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Atlas of Drosophila morphology: Wild-type and classical mutants. , (2013).</li><li>Yang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carnivory+in+the+larvae+of+Drosophila+melanogaster+and+other+Drosophila+species.">Carnivory in the larvae of Drosophila melanogaster and other Drosophila species.</a> Scientific Reports. 8, 15484 (2018).</li><li>Piper, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+holidic+medium+for+Drosophila+melanogaster.">A holidic medium for Drosophila melanogaster.</a> Nature Methods. 11, 100-105 (2014).</li><li>Shiraiwa, T., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proboscis+extension+response+(PER)+assay+in+Drosophila.">Proboscis extension response (PER) assay in Drosophila.</a> Journal of visualized experiments : JoVE. (3), e193 (2007).</li><li>Yang, C. -. H., Belawat, P., Hafen, E., Jan, L. Y., Jan, Y. -. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+egg-laying+site+selection+as+a+system+to+study+simple+decision-making+processes.">Drosophila egg-laying site selection as a system to study simple decision-making processes.</a> Science. 319 (5870), 1679-1683 (2008).</li><li>Yang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+homemade+tools+to+handle+fruit+flies-Drosophila+melanogaster.">Simple homemade tools to handle fruit flies-Drosophila melanogaster.</a> Journal Of Visualized Experiments: JoVE. (149), e59613 (2019).</li><li>Nichols, C. D., Becnel, J., Pandey, U. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+assay+Drosophila+behavior.">Methods to assay Drosophila behavior.</a> Journal of Visualized Experiments: JoVE. (61), e3795 (2012).</li><li>Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthetizing+Drosophila+for+behavioural+studies.">Anesthetizing Drosophila for behavioural studies.</a> Journal of Insect Physiology. 46 (4), 439-442 (2000).</li><li>. My not quite complete protective filter article Available from: <a target="_blank" href="https://www.lensrentals.com/blog/2017/06/the-comprehensive-ranking-of-the-major-uv-filters-on-the-market">https://www.lensrentals.com/blog/2017/06/the-comprehensive-ranking-of-the-major-uv-filters-on-the-market</a> (2017)</li><li>. UV and clear lens protection filters review Available from: <a target="_blank" href="https://www.the-digital-picture.com/Reviews/UV-and-Clear-Lens-Protection-Filters.aspx">https://www.the-digital-picture.com/Reviews/UV-and-Clear-Lens-Protection-Filters.aspx</a> (2013)</li><li>Shaw, P. J., Cirelli, C., Greenspan, R. J., Tononi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlates+of+sleep+and+waking+in+Drosophila+melanogaster.">Correlates of sleep and waking in Drosophila melanogaster.</a> Science. 287 (5459), 1834-1837 (2000).</li><li>Barradale, F., Sinha, K., Lebestky, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+Drosophila+grooming+behavior.">Quantification of Drosophila grooming behavior.</a> Journal of Visualized Experiments: JoVE. (125), e55231 (2017).</li><li>Szebenyi, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cleaning+behaviour+in+Drosophila+melanogaster.">Cleaning behaviour in Drosophila melanogaster.</a> Animal Behaviour. 17 (4), 641-651 (1969).</li><li>. Plexiglas : optical and transmission characteristics Available from: <a target="_blank" href="https://www.plexiglas.com/export/sites/plexiglas/.content/medias/downloads/sheet-docs/plexiglas-optical-and-transmission-characteristics.pdf">https://www.plexiglas.com/export/sites/plexiglas/.content/medias/downloads/sheet-docs/plexiglas-optical-and-transmission-characteristics.pdf</a> (2000)</li><li>. Calculation of the light reflection and transmission Available from: <a target="_blank" href="https://www.glassproperties.com/reflection/">https://www.glassproperties.com/reflection/</a> (2007)</li><li>Hunter, F., Biver, S., Fuqua, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Light science &amp; magic: an introduction to photographic Lighting. , (2015).</li><li>Hendricks, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rest+in+Drosophila+is+a+sleep-like+state.">Rest in Drosophila is a sleep-like state.</a> Neuron. 25 (1), 129-138 (2000).</li><li>Rieger, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fruit+fly+Drosophila+melangaster+favors+dim+light+and+times+its+activity+peaks+to+early+dawn+and+late+dusk.">The fruit fly Drosophila melangaster favors dim light and times its activity peaks to early dawn and late dusk.</a> Journal of Biological Rhythms. 22 (5), 387-399 (2007).</li></ol>

Light is at the very heart of photography and videography and is the decisive factor for obtaining high-quality images16. Here, two LED video lights with adjustable brightness and color temperature were used as illuminators, and a translucent material was selected to make the FBOC. The LED light panels on both sides provided enough brightness, and the translucent material softened and scattered light, eventually producing uniform, soft, and bright light to illuminate the flies in the FBOC, without...

Drosophila melanogaster is an outstanding model in biological research; however, it is a bad model for photography or videography, as it is too small for a camera or a camcorder and too large for a compound microscope1. Despite excellent research described in the literature, most studies have only provided blurred, unclear images, rather than clear and sharp photographs with clear detail that illustrate the fly behavior being described. Moreover, although fly behaviors have been extensively studied (e.g., courtship and fighting), most of these papers have used illustrations to explain their research to readers.
This paper describes a simple and economical approach. Using this method, not only the various behaviors of Drosophila can be observed, but also all the details that can be observed under a stereo zoom microscope can be recorded clearly and sharply. Of course, this method can also be used to record the morphology of flies, as when they enter a sleep or semi-sleep state, the stationary model allows the user to take a photograph or a stack of photographs with different focal planes to get an extended depth of field photo. These methods can be realized without complicated technology and expensive equipment or even superb manual skills.
The video component of this article shows videos of several typical behaviors of flies. The purpose of showing these videos is twofold: one is to let audiences know what can be captured and present the image quality obtained by using this method; the other is to let new students who are interested in Drosophila, but thus far have not had the opportunity to actually observe the behavior of flies understand the behavior of flies (such as courtship, fighting) through these clear videos rather than illustrations or blurred images.
Supplemental Video: Fruit Fly Behavior: <a href="https://www.jove.com/files/ftp_upload/62533/fly behavior.mp4" target="_blank">Please click here to download this file.</a>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>compact camera, Nikon P310</td>
			<td>Nikon</td>
			<td></td>
			<td>3-5x optical zoom, cam record 1080 P HD video</td>
		</tr>
		<tr>
			<td>ethylamine foam</td>
			<td></td>
			<td></td>
			<td>60 mm x 60 mm x 15 mm</td>
		</tr>
		<tr>
			<td>Food Blue No 1</td>
			<td></td>
			<td>CAS 3844-45-9</td>
			<td></td>
		</tr>
		<tr>
			<td>mini LED lights and transformer</td>
			<td>GODOX</td>
			<td>LED-P120</td>
			<td>have 5000-5600 K color temperature</td>
		</tr>
		<tr>
			<td>small container (e.g. bottle cap)</td>
			<td></td>
			<td></td>
			<td>about &#216; 15 mm x 20 mm</td>
		</tr>
		<tr>
			<td>UV / Clear filter</td>
			<td></td>
			<td></td>
			<td>high-quality UV/Clear filter with high transmittance, 30-40 mm</td>
		</tr>
		<tr>
			<td>zoom stereo microscope</td>
			<td></td>
			<td></td>
			<td>5-50x zoom</td>
		</tr>
	</tbody>
</table>

an adjustable high-definition imaging system for behavioral studies of drosophila adults

Drosophila melanogaster is a very powerful model in biological research, but a bad model for photography or videography. This paper describes a simple but effective method to observe and document the behavior or morphology of flies. Flies were placed in a translucent observation chamber c.a. &#216;15 x 5mm (no food inside) or &#216;15 x 12 mm (with an 8 mm-high piece of food inside). After covering with an ultraviolet (UV)/clear filter with high light transmittance, the chamber was placed under a 5-50x zoom stereo microscope, and mini light-emitting diode (LED) video lights were placed on both sides of the microscope to illuminate the chamber to obtain uniform, soft, bright, and nearly shadow-free light. Then, a compact digital camera with 3-5x optical zoom, which can record 1080 P high-definition or higher resolution video (at a frame rate of &#8805;30 fps), was connected to the eyepiece of microscope through a bracket, and photographs or videos were taken through the eyepiece. By adjusting the zoom knob of the zoom stereo microscope, it was very easy to track the flies and take panoramic or detailed close-up images as needed, while the camera recorded everything under the microscope. Because the flies can stay at any position in the chamber, they can be observed and recorded from all directions. The photographs or videos taken are of good image quality. This method can be used both for scientific research and teaching.

Shoot through a UV filter for clear and sharp images 
Perform a simple experiment to observe the difference between a UV filter and ordinary glass in the laboratory. Take a fly culture vial, remove the stopper, place it under a stereo dissecting microscope, and cover it (alternately) with a UV filter and a Petri dish lid. The photographs taken in these two cases are shown in Figure 5. The photograph taken through the UV filter is clear and sharp, very similar to the pho...

Watch this Scientific Journal Video about An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults at JoVE.com

An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults

This protocol describes how to make a simple adult Drosophila behavior observation chamber, and how to take high-definition photographs/videos of the morphology or behavior of different types of fruit flies in the observation chamber through relatively simple and affordable methods.

An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults

Drosophila melanogaster is a very powerful model in biological research, but a bad model for photography or videography. This paper describes a simple but ...

Due to its awkward size, Drosophila is difficult to image. This method demonstrates imaging of the behavior and morphology of flies through use of a stereo microscope. This protocol can be used both for scientific research and teaching.Demonstrating the procedure will be Ms.Li Tong and Ms.Weng Yujia, undergraduates doing research training under my guidance. To begin, make a fly behavior observation chamber, or FBOC, using a translucent plastic bottle cap of about 17 by 22 millimeters or by cutting a section of 17 millimeters from the thick end of a five milliliter pipette tip. Pour 1%agar into the FBOC to adjust its depth.If food needs to be placed in the FBOC, pour the agar to obtain a depth of 12 millimeters. If food does not need to be placed in the FBOC, pour the agar to a depth of five millimeters to track the whereabouts of fruit flies more easily. If using a pipette tip to make an FBOC, place the cutoff pipette tip section in a 35 or a 60 millimeter Petri dish.Pour the 1%agar gel into the Petri dish to a thickness of about five millimeter and wait for the agar to solidify and seal the bottom of the FBOC. Then pour the agar gel into the FBOC to the desired thickness. Make an FBOC base by boring a 10 millimeter deep hole in the center of a piece of ethylamine foam sheet with the same diameter as the FBOC.Insert the FBOC into the hole. Make fly food if necessary with yeast medium, artificial diet, or pure sucrose or glucose, depending on the purpose of the observation. To visually determine whether flies are feeding, add food dyes to the food to a final concentration of 12.5 milligrams per 100 milliliters.Pour the prepared food into a Petri dish to a height of eight millimeters. After solidification, place food on a rectangular drawn on paper and cut the food accordingly and place it on a piece of plastic and cut into a quadrangular pyramid or quadrangular frustum pyramid to allow recording of fly behavior from different angles as the flies land on the food. Cut a piece of plastic and use tweezers to place the plastic and food in the center of the FBOC.Pour 1%agar gel into a clean empty bottle to a thickness of 1-2 centimeters and place it at room temperature for 1-2 hours. Transfer flies to the bottle and place it at 25 degrees Celsius for 36 hours or more. Chill and inactivate the flies in crushed ice, sort them on an ice pack and transfer them to the FBOC as previously described.After transferring the flies to the FBOC, cover it with 30-40 millimeters of UV or clear filter for the camera to form an FBOC complex. Place the FBOC complex under the stereo microscope for observation. Illuminate the FBOC.Mount mini LED video lights to flash hot shoe mount stands and place the lights on the left and right sides of the FBOC. Turn on the lights and set the color temperature to 5, 000 to 5, 600 Kelvin and the brightness to 100%Turn on the LED video lights and adjust the stereo zoom microscope until the edge of the FBOC can be clearly seen with the naked eye. Move the FBOC to the center of the field of view.Attach the clamp of the universal telescope digital camera adapter to an eyepiece of the stereo microscope. Then attach a compact digital camera to the adapter securely by alternately turning the camera mounting screw and camera fixing screw. Turn on the digital camera and turn the horizontal/vertical fine-tuning knobs until the FBOC edge clearly appears in the center of the bright circular field of view on the camera's LCD screen.Rotate the mode dial to aperture priority auto-mode. Press focus mode on the multi-selector, choose macro close-up and then press OK button. Move the zoom switch from the wide angle end to the telephoto end and zoom into the circular image until its central portion fills the full LCD screen.Press the movie-record button and start recording the fly behavior. Turn the focus knob of the microscope until the flies in the FBOC are clearly visible. Choose the fly behavior of interest for observation or video recording.Turn the zoom knob to zoom in and out to achieve the desired magnification for observation or video recording. The representative photograph taken through the UV filter is clear and sharp, very similar to the photograph taken when the culture vial is not covered. The quality of the photo taken through the glass of the Petri dish is very poor and partly blurred.A photograph taken from the video recording showing the details of each part of the fly's body is shown here. Because the camera is connected to a zoom stereo microscope, it is very easy to shoot from panoramic to close-up shots using the zoom system. Using this protocol, fly behavior can be observed and documented for multiple viewing angles.For example, the female fly is constantly rubbing the ovipositor with her hind legs during the process of laying eggs. This detail of egg-laying behavior cannot be seen clearly from the side. Using this simple and economical method, we can take a panoramic or close-up of the behavior and morphology of flies and obtain a good quality video or photo.

Research

JoVE Journal

Behavior

Behavioral Assessment of the Aging Mouse Vestibular System

Age related decline in balance performance is associated with deteriorating muscle strength, motor coordination and vestibular function. While a number of ...

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

Using a Real-Time Locating System to Measure Walking Activity Associated with Wandering Behaviors Among Institutionalized Older Adults

A real-time locating system (RTLS) can be used to track the walking activity of institutionalized older adults in long-term care who are at risk for ...

Behavioral Tracking and Neuromast Imaging of Mexican Cavefish

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among ...

Lexical Decision Task for Studying Written Word Recognition in Adults with and without Dementia or Mild Cognitive Impairment

Older adults are slower at recognizing visual objects than younger adults. The same is true for recognizing that a letter string is a real word. People ...

An Experimental Paradigm for Measuring the Effects of Ageing on Sentence Processing

Previous studies have found that older adults have greater difficulties in processing syntactically complex sentences than younger adults. However, the ...

Flypub To Study Ethanol Induced Behavioral Disinhibition and Sensitization

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the ...