Calcium Imaging
Calcium imaging is a scientific technique usually carried out in research which is designed to show the calcium (Ca2+) status of a cell, tissue or medium. Calcium imaging techniques take advantage of so-called calcium indicators, fluorescent molecules that can respond to the binding of Ca2+ ions by changing their fluorescence properties.
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- Cameras
- Light Source – Monochromator
- Software
- Systems
Cameras
Evolve EMCCD Cameras
The Evolve® is a high-resolution, back-illuminated EMCCD providing high sensitivity for the lowest light applications.
With proprietary Teledyne technology throughout, the Evolve provides a new standard in EMCCD performance and reliability. For single molecule imaging, TIRF microscopy, and more, achieve the best imaging performance when every photon counts.
- >95% quantum efficiency
- 10, 13 and 16 µm pixel size options
- Ultra-high-speed readout
- Highly stabilized deep cooling
- Featuring Teledyne EMCCD technology
Light Source – Monochromator
Cairn Monochromator
- High throughput F/2 corrected optic
- Integral fast electronic shutter on all models
The Optoscan monochromator from Cairn Research is the only instrument of its type to offer control of both centre-wavelength and bandwidth with millisecond time resolution. Combined with Cairn signal processing modules and optical hardware it forms the heart of a powerful and flexible microfluorescence system. Alternatively it can be readily integrated into commercial imaging systems, or custom controlled for bespoke applications.
This instrument has been designed specifically with real-time biological fluorescence measurements in mind, but has evolved into a truly versatile laboratory tool. If your application requires fast, flexible and automated illumination control then the Cairn Optoscan may well be the instrument of choice.
Other commercial monochromators include the facility to change bandwidth between experiments using a manual control. This is sufficient for many purposes, but is not ideal for complex protocols especially those involving multiple fluorescence markers. Only by controlling bandwidth in real-time during experiments can each excitation wavelength be optimised independently. This means that where Stoke’s shifts are large or where fluorescence intensities are weak then a relatively large bandwidth can be selected to maximise signal-to-noise. Conversely if there is a small Stoke’s shift or if the fluorescence intensity is high then the slit width can be minimised to reduce bleed through and optimise dynamic range.
Software
MetaFluor Fluorescence Ratio Imaging Software
MetaFluor® Fluorescence Ratio Imaging Software is the industry standard for monitoring fluorescence ratio imaging applications,
bringing microscopists closer to rapid cellular responses inherent to live cells.
MetaFluor Fluorescence Ratio Imaging
Software is designed for dual-wavelength intracellular ion measurements. The system provides simultaneous display of the raw
data; ratio image; graphs of intensities, ratios, and ion concentrations (e.g., calcium, sodium, potassium, chloride); and a
non-ratiometric image such as a brightfield or phase-contrast image. Two different ratiometric indicators can be imaged and
measured simultaneously to provide greater insight to ion exchange and intracellular function regardless of dye loading
concentrations, conditions, or emission intensities.
Toolbars, menus, wizards, and dialog boxes help move you through
the image processing steps quickly. Features such as multiple image windows, flexible device control, synchronization and timing,
and journals allow for automated image acquisition and analysis unlike any other system.
With MetaFluor Fluorescence
Software, you customize the set-up once, then let the experiment run by itself. You are able to collect a large amount of data
online and process it with either MetaFluor Software or independently using an analysis-only software
version.
MetaFluor Fluorescence Ratio Imaging Software features:
- Real time processing significantly reduces random image noise by averaging up to 256 images per time point. Background subtraction is also used to improve accuracy by correcting for stray light, camera noise, and auto-fluorescence.
- Calibrations of intracellular ion concentrations are obtained by using the various calibration options offered: the Grynkiewicz equation (Grynkiewicz et al., 1985) and titration equation for both in situ and in vitro experiments. These calibrations can then be stored for future use.
- Region analysis can be generated automatically or manually to monitor intensity, ratio value, or ion concentration. Measurements are then made simultaneously on all the regions of interest and update continuously on a scrolling graph, allowing you to follow dynamic changes as they occur in your living samples.
- Interactive graphs give flexibility in the presentation of your experiment’s data. MetaFluor Software enables you to click on graph traces to display a readout of the time and data value for the region nearest to the click.
- Event mark function is useful to record when drugs or solutions are added, experimental conditions changed, triggers are received or sent, or other events occurred. You have the option to associate a timer and an alarm bell to each event. Additionally, for perfused samples, ambient conditions can be logged and tracked.
- Journals allow you to create functions to simplify system operations, automate acquisition and device control, set variables, and sequence events. The Auto-Execute Journal command allows you to automatically run journals at specific points in the acquisition cycle.
Systems
OASIS In Vivo Platform – Modular Macroscope and Microscope
Optical Activation, Stimulation & Imaging System
- Translates and Rotates for Live Animal Research
- Simultaneously Image & Deliver Wide-field and/or Pattern Illumination
- Macro or Microscopic Optical Assembly
- Modular Platform Tailors to any Experimental Application
- Excellent System for:
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- In Vivo Optogenetics
- Photostimulation
- Photoactivation
Mightex’s Optical Activation, Stimulation and Imaging System (OASIS) is designed specifically to meet the growing demand to
conduct experiments on live animals, when system flexibility is a primary consideration and a large Field of View (FoV) is
usually required. In Optogenetics, for example, in vivo patterned illumination on rodents, monkeys and other animals is the next
step towards the ultimate goal of being able to record and control the activity of each neuron in the brain of behaving animals
at a single cell and ms resolution. To achieve patterned illumination for in vivo applications, the OASIS features Mightex’s
market-leading Polygon400 patterned Illuminator to deliver light to a specified area of the specimen while leaving neighboring
areas untouched.
Whereas traditional microscopes are a fixed system with limited flexibility, OASIS is designed to
provide excellent reconfigurability/flexibility and can be configured to work around a fixed specimen such as a sedated live
animal. A heavy-duty precision XYZ stage is mounted on the support structure and the OASIS optical assembly is in turn mounted on
the XYZ stage. The XYZ stage controls the lateral motion as well as focus of the system. A pan/tilt mechanism add-on is also
available to orient the OASIS relative to the live animal. Motorized versions of both the XYZ stage and tilt mechanism are
available.
OASIS Implant – Deep-Brain Imaging & Stimulation System for Freely Behaving Animals
Key Features
- In vivo all-optical recording and manipulatiion of neural circuit activity;
- Sub-cellular resolution, high-quality imaging for quantitative analysis;
- Multi-wavelength illumination;
- Patterned stimulation with sub-cellular targeting capability;
- Super-light head mounted fixture, 0.3g;
- Modular system design, reconfigurable for different applications.
Mightex’s OASIS Implant is a ground-breaking microendoscopic system that is capable of doing deep-brain simultaneous all-optical recording and manipulation of neural circuit activity with cellular resolution, on freely behaving animals. Compatible with high-sensitivity research-grade cameras, OASIS Implant enables researchers to acquire far superior quality deep-brain images & videos, compared to the relatively low-quality head-mounted cameras that have recently been introduced into the market. In addition, with multiple illumination ports, OASIS Implant is equipped with both wide-field and patterned illumination capabilities (when combined with Mightex’s market-leading Polygon400) and, moreover, it is also capable of doing multi-wavelength imaging/stimulation which cannot be done with the existing head-mounted cameras. More importantly, since both the deep-brain imaging and the deep-brain stimulation are delivered through an Imaging Fiber bundle, the overall weight of the head-mounted fixture is as little as 0.3g (compared to approximately 2g with a head-mounted camera), making OASIS Implant an ideal research tool for in vivo live animal experiments.
In addition to many other applications, OASIS Implant enables bioscientists to do the following:
- Deep brain calcium imaging in freely behaving animals;
- Deep brain patterned optogenetic stimulation with single-cell or sub-cellular resolution;
- Simultaneous deep brain stimulation and imaging.