Low-cost conversion of the Polaroid MD-4 land camera to a digital gel documentation system

A simple, inexpensive design is presented for the rapid conversion of the popular MD-4 Polaroid land camera to a high quality digital gel documentation system. Images of ethidium bromide stained DNA gels captured using the digital system were compared to images captured on Polaroid instant film. Resolution and sensitivity were enhanced using the digital system. In addition to the low cost and superior image quality of the digital system, there is also the added convenience of real-time image viewing through the swivel LCD of the digital camera, wide flexibility of gel sizes, accurate automatic focusing, variable image resolution, and consistent ease of use and quality. Images can be directly imported to a computer by using the USB port on the digital camera, further enhancing the potential of the digital system for documentation, analysis, and archiving. The system is appropriate for use as a start-up gel documentation system and for routine gel analysis.     

Imaging technologies for the detection of multiple stains in proteomics

Laser-based scanners and charge-coupled device (CCD) camera systems are evolving to have greater functional capabilities for capturing images from a range of staining technologies used in gel electrophoresis and electroblotting. Digitizing Coomassie Brilliant Blue (CBB) stained gels and silver stained gels has now become possible using a laser-based gel scanner, the FLA-5000 fluorescent image analyzer system. Also, a simultaneous dual fluorescent imaging function has been incorporated into the FLA-5000 system, utilizing dichroic mirrors with both the optical system and the emission filter. In the workflow of routine proteomics research, the relationship between SYPRO dye staining and fluorescent detection using the FLA-5000 system have become symbiotic. Additionally in many cases, subsequent staining of the gel with CBB is useful for future research, and thus imaging instruments should be able to handle both staining formats. Digitizing the CBB stained gel can now be easily performed by the FLA-5000 fluorescent image analyzer system using a fluorescent board as an epi-illumination background. A cooled CCD camera system has the potential of imaging not only chemiluminescent membranes but also digitizing molecular weight markers and fluorescent detection of SYPRO dye-stained gels. With Multi Gauge software version 2.0 it is now a simple task to combine two images into one, as commonly required in dual detection experiments. The LAS-3000 system was designed to capture chemiluminescent images and to digitize the images automatically. Thus, new capabilities added to gel imaging systems make them capable of detecting and displaying multiple signals more conveniently.     

Comparison of whole-cell protein electrophoretic profiles of Haemophilus influenzae: implementation of a microcomputer mainframe linked system and description of a new similarity coefficient

A microcomputer mainframe linked system is described which allows video camera data capture and storage of one-dimensional whole-cell protein electrophoresis gel images, processing of normalized traces to produce a similarity matrix, and analysis of the matrix using the commercial cluster analysis program CLUSTAN. A new similarity coefficient is introduced which takes into account both band position and intensity. Forty-five strains of Haemophilus influenzae, including the eight biotypes and six serotypes, were analyzed using this system. Results demonstrated groupings which are consistent with known genetic relationships.     

A reconfigurable real-time compressive-sampling camera for biological applications

Many applications in biology, such as long-term functional imaging of neural and cardiac systems, require continuous high-speed imaging. This is typically not possible, however, using commercially available systems. The frame rate and the recording time of high-speed cameras are limited by the digitization rate and the capacity of on-camera memory. Further restrictions are often imposed by the limited bandwidth of the data link to the host computer. Even if the system bandwidth is not a limiting factor, continuous high-speed acquisition results in very large volumes of data that are difficult to handle, particularly when real-time analysis is required. In response to this issue many cameras allow a predetermined, rectangular region of interest (ROI) to be sampled, however this approach lacks flexibility and is blind to the image region outside of the ROI. We have addressed this problem by building a camera system using a randomly-addressable CMOS sensor. The camera has a low bandwidth, but is able to capture continuous high-speed images of an arbitrarily defined ROI, using most of the available bandwidth, while simultaneously acquiring low-speed, full frame images using the remaining bandwidth. In addition, the camera is able to use the full-frame information to recalculate the positions of targets and update the high-speed ROIs without interrupting acquisition. In this way the camera is capable of imaging moving targets at high-speed while simultaneously imaging the whole frame at a lower speed. We have used this camera system to monitor the heartbeat and blood cell flow of a water flea (Daphnia) at frame rates in excess of 1500 fps.     

The use of continuous monitoring and computer-assisted image analysis for the histochemical quantification of hexokinase activity

The effect of endogenous 6-phosphogluconate dehydrogenase (6PGDH) on the histochemical quantification of low-Km hexokinase activity in rat submandibular salivary glands has been investigated using a Seescan Solitaire Plus image analysis system and a modified black and white Newvicon TV camera. Absorbance readings of neutral density filters were close to their known absorbance values. A significant correlation was found between the absorbance of Nitro BT reduction products within sections, with and without the use of a 588 nm interference filter. Furthermore, absorbance readings obtained from 8 microns-thick sections were 1.92 and 2.22 times greater than values obtained from 4 microns-thick sections using 'white' and 588 nm light respectively. The level of background illumination was not critical for absorbance measurements provided it was below the level that saturated the Newvicon camera and was not changed between background and image capture. The greatest variations in absorbance readings on tissue sections were associated with changes in zoom and objective combinations. Our studies indicate that this relatively low cost image analysis system can give reproducible absorbance readings from various structures defined in digitized, captured images of tissue sections. Results from continuous monitoring studies indicated that the high levels of 6PGDH activity in excretory ducts caused a 1.67-fold overestimation of hexokinase activity as assessed by absorbance readings performed throughout a 22-minute reaction period. By contrast, overestimation of hexokinase activity in salivary gland acini only became apparent after 8 min incubation with a 1.4-fold overestimation being seen at 22 min. This difference appears to reflect the relatively low hexokinase and 6PGDH activities present in acini compared with excretory ducts.     

Accuracy and repeatability of joint angles measured using a single camera markerless motion capture system

Markerless motion capture systems have developed in an effort to evaluate human movement in a natural setting. However, the accuracy and reliability of these systems remain understudied. Therefore, the goals of this study were to quantify the accuracy and repeatability of joint angles using a single camera markerless motion capture system and to compare the markerless system performance with that of a marker-based system. A jig was placed in multiple static postures with marker trajectories collected using a ten camera motion analysis system. Depth and color image data were simultaneously collected from a single Microsoft Kinect camera, which was subsequently used to calculate virtual marker trajectories. A digital inclinometer provided a measure of ground-truth for sagittal and frontal plane joint angles. Joint angles were calculated with marker data from both motion capture systems using successive body-fixed rotations. The sagittal and frontal plane joint angles calculated from the marker-based and markerless system agreed with inclinometer measurements by <0.5°. The systems agreed with each other by <0.5° for sagittal and frontal plane joint angles and <2° for transverse plane rotation. Both systems showed a coefficient of reliability <0.5° for all angles. These results illustrate the feasibility of a single camera markerless motion capture system to accurately measure lower extremity kinematics and provide a first step in using this technology to discern clinically relevant differences in the joint kinematics of patient populations.     

Screening algal mutant colonies with altered thylakoid electrochemical gradient through fluorescence and delayed luminescence digital imaging

A digital imaging instrument intended to monitor fluorescence and delayed luminescence of algal colonies grown on Petri plates is described. The system includes light-emitting diodes, a cooled line transfer charge-coupled device (CCD) camera, and a personal computer. Software developments were made to capture pictures and kinetics in real time during illuminations. This instrument makes the fluorescence induction kinetics of individual colonies readily accessible with a good time resolution. It offers a great refinement for screening colonies deficient in photosynthetic electron transfer thanks to appropriate computed fluorescence images. The high sensitivity of the instrument allows it to capture fluorescence images under non-actinic illuminations, and for the first time, delayed luminescence images, opening thus the way to screening mutants that have altered thylakoid electrochemical gradients.     

The Gel Electrophoresis Markup Language (GelML) from the Proteomics Standards Initiative

The Human Proteome Organisation's Proteomics Standards Initiative has developed the GelML (gel electrophoresis markup language) data exchange format for representing gel electrophoresis experiments performed in proteomics investigations. The format closely follows the reporting guidelines for gel electrophoresis, which are part of the Minimum Information About a Proteomics Experiment (MIAPE) set of modules. GelML supports the capture of metadata (such as experimental protocols) and data (such as gel images) resulting from gel electrophoresis so that laboratories can be compliant with the MIAPE Gel Electrophoresis guidelines, while allowing such data sets to be exchanged or downloaded from public repositories. The format is sufficiently flexible to capture data from a broad range of experimental processes, and complements other PSI formats for MS data and the results of protein and peptide identifications to capture entire gel‐based proteome workflows. GelML has resulted from the open standardisation process of PSI consisting of both public consultation and anonymous review of the specifications.     

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While these systems can provide a wealth of behavioral information, the cost and complexity of these systems can be prohibitive for many labs. We have developed a low-cost assay for measuring locomotive behavior and seizure movement in D. melanogaster. The system uses a web-cam to capture images that can be processed using a combination of inexpensive and free software to track the distance moved, the average velocity of movement and the duration of movement during a specified time-span. To demonstrate the utility of this system, we examined a group of D. melanogaster mutants, the Bang-sensitive (BS) paralytics, which are 3-10 times more susceptible to seizure-like activity (SLA) than wild type flies. Using this novel system, we were able to detect that the BS mutant bang senseless (bss) exhibits lower levels of exploratory locomotion in a novel environment than wild type flies. In addition, the system was used to identify that the drug metformin, which is commonly used to treat type II diabetes, reduces the intensity of SLA in the BS mutants.     

The selfie trap: A novel camera trap design for accurate small mammal identification

Camera traps are a popular tool for monitoring wildlife though they can fail to capture enough morphological detail for accurate small mammal species identification. Camera trapping small mammals is often limited by the inability of camera models to: (i) record at close distances; and (ii) provide standardised photos. This study aims to provide a camera trapping method that captures standardised images of the faces of small mammals for accurate species identification, with further potential for individual identification. A novel camera trap design coined the ‘selfie trap’ was developed. The selfie trap is a camera contained within an enclosed PVC pipe with a modified lens that produces standardised close images of small mammal species encountered in this study, including: Brown Antechinus (Antechinus stuartii), Bush Rat (Rattus fuscipes) and Sugar Glider (Petaurus breviceps). Individual identification was tested on the common arboreal Sugar Glider. Five individual Sugar Gliders were identified based on unique head stripe pelage. The selfie trap is an accurate camera trapping method for capturing detailed and standardised images of small mammal species. The design described may be useful for wildlife management as a reliable method for surveying small mammal species. However, intraspecies individual identification using the selfie trap requires further testing.     

The effect of attractant lures in camera trapping: a case study of population estimates for the Iberian lynx (Lynx pardinus)

Capture–recapture analysis of camera trap data is a conventional method to estimate the abundance of free-ranging wild felids. Due to notorious low detection rates of felids, it is important to increase the detection probability during sampling. In this study, we report the effectiveness of attractants as a tool for improving the efficiency of camera trap sampling in abundance estimation of Iberian lynx. We developed a grid system of camera stations in which stations with and without attractant lures were spatially alternated across known Iberian lynx habitat. Of the ten individuals identified, five were detected at stations with no attractant (blind sets), and nine, at the lured stations. Thirty-eight percent of blind set station’s independent captures and 10?% of lured station’s independent captures resulted in photographs unsuitable for correct individual identification. The total capture probability at lured stations was higher than that obtained at blind set stations. The estimates obtained with blind set cameras underestimated the number of lynxes compared to lured cameras. In our study, it appears that the use of lures increased the efficiency of trail camera captures and, therefore, the accuracy of capture–recapture analysis. The observed failure to detect known individuals at blind set camera stations may violate capture–recapture assumptions and bias abundance estimates.     

Trapping Elusive Cats: Using Intensive Camera Trapping to Estimate the Density of a Rare African Felid

Camera trapping studies have become increasingly popular to produce population estimates of individually recognisable mammals. Yet, monitoring techniques for rare species which occur at extremely low densities are lacking. Additionally, species which have unpredictable movements may make obtaining reliable population estimates challenging due to low detectability. Our study explores the effectiveness of intensive camera trapping for estimating cheetah (Acinonyx jubatus) numbers. Using both a more traditional, systematic grid approach and pre-determined, targeted sites for camera placement, the cheetah population of the Northern Tuli Game Reserve, Botswana was sampled between December 2012 and October 2013. Placement of cameras in a regular grid pattern yielded very few (n = 9) cheetah images and these were insufficient to estimate cheetah density. However, pre-selected cheetah scent-marking posts provided 53 images of seven adult cheetahs (0.61 ± 0.18 cheetahs/100km²). While increasing the length of the camera trapping survey from 90 to 130 days increased the total number of cheetah images obtained (from 53 to 200), no new individuals were recorded and the estimated population density remained stable. Thus, our study demonstrates that targeted camera placement (irrespective of survey duration) is necessary for reliably assessing cheetah densities where populations are naturally very low or dominated by transient individuals. Significantly our approach can easily be applied to other rare predator species.     

Photoduplication and fluorescence measurement in polyacrylamide gel electrophoresis.

A simple method of photoduplication of gel electrophoresis, visualized with fluorescent reagents, is described. The procedure is convenient and rapid and requires no camera or expensive equipment. Using electrophoresis duplicating paper (Kodak), positive prints suitable for documentation or publication may be obtained. With usual photographic paper, negative prints may be obtained, allowing reliable measurement by scanning. The technique may be applied to protein or nucleic acid electrophoresis.     

Unmanned Aerial Vehicles (UAVs) for Surveying Marine Fauna: A Dugong Case Study

Aerial surveys of marine mammals are routinely conducted to assess and monitor species’ habitat use and population status. In Australia, dugongs (Dugong dugon) are regularly surveyed and long-term datasets have formed the basis for defining habitat of high conservation value and risk assessments of human impacts. Unmanned aerial vehicles (UAVs) may facilitate more accurate, human-risk free, and cheaper aerial surveys. We undertook the first Australian UAV survey trial in Shark Bay, western Australia. We conducted seven flights of the ScanEagle UAV, mounted with a digital SLR camera payload. During each flight, ten transects covering a 1.3 km² area frequently used by dugongs, were flown at 500, 750 and 1000 ft. Image (photograph) capture was controlled via the Ground Control Station and the capture rate was scheduled to achieve a prescribed 10% overlap between images along transect lines. Images were manually reviewed post hoc for animals and scored according to sun glitter, Beaufort Sea state and turbidity. We captured 6243 images, 627 containing dugongs. We also identified whales, dolphins, turtles and a range of other fauna. Of all possible dugong sightings, 95% (CI = 90%, 98%) were subjectively classed as ‘certain’ (unmistakably dugongs). Neither our dugong sighting rate, nor our ability to identify dugongs with certainty, were affected by UAV altitude. Turbidity was the only environmental variable significantly affecting the dugong sighting rate. Our results suggest that UAV systems may not be limited by sea state conditions in the same manner as sightings from manned surveys. The overlap between images proved valuable for detecting animals that were masked by sun glitter in the corners of images, and identifying animals initially captured at awkward body angles. This initial trial of a basic camera system has successfully demonstrated that the ScanEagle UAV has great potential as a tool for marine mammal aerial surveys.     

Simultaneously Capturing Real-time Images in Two Emission Channels Using a Dual Camera Emission Splitting System: Applications to Cell Adhesion

Multi-color immunofluorescence microscopy to detect specific molecules in the cell membrane can be coupled with parallel plate flow chamber assays to investigate mechanisms governing cell adhesion under dynamic flow conditions. For instance, cancer cells labeled with multiple fluorophores can be perfused over a potentially reactive substrate to model mechanisms of cancer metastasis. However, multi-channel single camera systems and color cameras exhibit shortcomings in image acquisition for real-time live cell analysis. To overcome these limitations, we used a dual camera emission splitting system to simultaneously capture real-time image sequences of fluorescently labeled cells in the flow chamber. Dual camera emission splitting systems filter defined wavelength ranges into two monochrome CCD cameras, thereby simultaneously capturing two spatially identical but fluorophore-specific images. Subsequently, psuedocolored one-channel images are combined into a single real-time merged sequence that can reveal multiple target molecules on cells moving rapidly across a region of interest.     

Live Imaging of Dense-core Vesicles in Primary Cultured Hippocampal Neurons

Observing and characterizing dynamic cellular processes can yield important information about cellular activity that cannot be gained from static images. Vital fluorescent probes, particularly green fluorescent protein (GFP) have revolutionized cell biology stemming from the ability to label specific intracellular compartments and cellular structures. For example, the live imaging of GFP (and its spectral variants) chimeras have allowed for a dynamic analysis of the cytoskeleton, organelle transport, and membrane dynamics in a multitude of organisms and cell types [1-3]. Although live imaging has become prevalent, this approach still poses many technical challenges, particularly in primary cultured neurons. One challenge is the expression of GFP-tagged proteins in post-mitotic neurons; the other is the ability to capture fluorescent images while minimizing phototoxicity, photobleaching, and maintaining general cell health. Here we provide a protocol that describes a lipid-based transfection method that yields a relatively low transfection rate (~0.5%), however is ideal for the imaging of fully polarized neurons. A low transfection rate is essential so that single axons and dendrites can be characterized as to their orientation to the cell body to confirm directionality of transport, i.e., anterograde v. retrograde. Our approach to imaging GFP expressing neurons relies on a standard wide-field fluorescent microscope outfitted with a CCD camera, image capture software, and a heated imaging chamber. We have imaged a wide variety of organelles or structures, for example, dense-core vesicles, mitochondria, growth cones, and actin without any special optics or excitation requirements other than a fluorescent light source. Additionally, spectrally-distinct, fluorescently labeled proteins, e.g., GFP and dsRed-tagged proteins, can be visualized near simultaneously to characterize co-transport or other coordinated cellular events. The imaging approach described here is flexible for a variety of imaging applications and can be adopted by a laboratory for relatively little cost provided a microscope is available.     

An automatic method for removing empty camera trap images using ensemble learning

Camera traps often produce massive images, and empty images that do not contain animals are usually overwhelming. Deep learning is a machine‐learning algorithm and widely used to identify empty camera trap images automatically. Existing methods with high accuracy are based on millions of training samples (images) and require a lot of time and personnel costs to label the training samples manually. Reducing the number of training samples can save the cost of manually labeling images. However, the deep learning models based on a small dataset produce a large omission error of animal images that many animal images tend to be identified as empty images, which may lead to loss of the opportunities of discovering and observing species. Therefore, it is still a challenge to build the DCNN model with small errors on a small dataset. Using deep convolutional neural networks and a small‐size dataset, we proposed an ensemble learning approach based on conservative strategies to identify and remove empty images automatically. Furthermore, we proposed three automatic identifying schemes of empty images for users who accept different omission errors of animal images. Our experimental results showed that these three schemes automatically identified and removed 50.78%, 58.48%, and 77.51% of the empty images in the dataset when the omission errors were 0.70%, 1.13%, and 2.54%, respectively. The analysis showed that using our scheme to automatically identify empty images did not omit species information. It only slightly changed the frequency of species occurrence. When only a small dataset was available, our approach provided an alternative to users to automatically identify and remove empty images, which can significantly reduce the time and personnel costs required to manually remove empty images. The cost savings were comparable to the percentage of empty images removed by models.     

Automatic control of a robot camera for broadcasting based on cameramen's techniques and subjective evaluation and analysis of reproduced images

With the goal of achieving an intelligent robot camera system that can take dynamic images automatically through humanlike, natural camera work, we analyzed how images were shot, subjectively evaluated reproduced images, and examined effects of camerawork, using camera control technique as a parameter. It was found that (1) A high evaluation is obtained when human-based data are used for the position adjusting velocity curve of the target; (2) Evaluation scores are relatively high for images taken with feedback-feedforward camera control method for target movement in one direction; (3) Keeping the target within the image area using the control method that imitates human camera handling becomes increasingly difficult when the target changes both direction and velocity and becomes bigger and faster, and (4) The mechanical feedback method can cope with rapid changes in the target's direction and velocity, constantly keeping the target within the image area, though the viewer finds the image rather mechanical as opposed to humanlike.     

基于移动终端的稻田飞虱调查方法

【目的】建立一种基于移动终端的稻田飞虱调查方法,以减轻测报人员劳动强度,提高稻田飞虱调查的客观性,实现稻飞虱调查结果可追溯。【方法】利用Android相机、可伸缩手持杆和装载控制相机APP的Android手机研制了稻田飞虱图像采集仪。在Android开发环境下,利用socket通信和视频编码等技术,实现Android相机的视频采集与编码模块、视频传输模块和相机命令控制模块等。利用Android NDK开发和Java web等技术,实现手机端的视频预览模块、手机控制模块、图像上传模块等。相机实时拍摄的视频将压缩成H.264格式,通过RTSP/RTP协议控制其传输至手机端。手机端通过解压缩,实现实时预览相机所拍摄的视频,并控制相机拍摄水稻茎基部飞虱图像,同时将图像传输到手机端。稻飞虱识别算法部署在云服务器上。手机端可选择稻飞虱图像上传至云服务器,云服务器运行稻飞虱自动识别算法,结果返回至手机端。【结果】基于移动终端的稻田飞虱调查方法利用手机可以实时预览相机拍摄的水稻茎基部飞虱画面,控制相机拍照。云服务器上稻飞虱自动识别算法对图像中的飞虱平均检测率为86.9%,虚警率为11.2%;对稻飞虱各虫态平均检测率为81.7%,虚警率为16.6%。【结论】基于移动终端的稻田飞虱调查方法可以便捷地采集到水稻茎基部飞虱图像,实现稻田飞虱不同虫态的识别与计数。该方法可大大减轻测报人员的劳动量,避免稻飞虱田间调查的主观性,实现稻飞虱田间调查的可追溯。     

Method for testing motion analysis laboratory measurement systems

This paper proposes a method for comparing data from accelerometers, optical based 3D motion capture systems, and force platforms (FPs) in the context of spatial and temporal differences. Testing method is based on the motion laboratory accreditation test (MLAT), which can be used to test FP and camera based motion capture components of a motion analysis laboratory. This study extends MLAT to include accelerometer data. Accelerometers were attached to a device similar to the MLAT rod. The elevation of the rod from the plane of the floor is computed and compared with the force platform vector orientation and the rod orientation obtained by optical motion capture system. Orientation of the test device is achieved by forming nonlinear equation group, which describes the components of the measured accelerations. Solution for this equation group is estimated by using the Gauss-Newton method. This expanded MLAT procedure can be used in the laboratory setting were either FP, camera based motion capture, or any other motion capture system is used along with accelerometer measurements.