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Multi-Wavelength Imaging

Multi-Wavelength Imaging
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Multi-Wavelength Imaging

Product catalog summary
Overview
The document provides an in-depth analysis of Andor's solutions for simultaneous multi-wavelength imaging in fluorescence microscopy. It focuses on products such as Optosplit II, TuCam, and Optosplit III, which facilitate capturing images at multiple wavelengths using single or dual camera setups.
Optosplit II
The Optosplit II is an image splitter designed for dual emission imaging, allowing a single camera to capture images at two different wavelengths simultaneously. It is ideal for applications like FRET and ratiometric ion imaging, featuring variable path separation, interchangeable filters, and a compact design.
TuCam
The TuCam is a dual-camera adapter that supports simultaneous imaging from two cameras. It is compatible with various camera types and offers a large aperture, high transmission, and low distortion, making it suitable for dual wavelength super resolution and TIRF microscopy.
Optosplit III
The Optosplit III is a three-way image splitter that enables a single camera to capture images at up to three different wavelengths. It is designed for applications requiring simultaneous acquisition of multiple emission bands, supporting unity magnification and precise image positioning.
Key Applications
  • Real-time multi-color imaging
  • Co-localization studies
  • Fluorescence Resonance Energy Transfer (FRET)
  • Ratiometric imaging
  • Super resolution imaging
  • Calcium flux/ion signaling
Recommended Software
Several software packages are recommended for simultaneous dual camera acquisition, offering functionalities to merge and analyze data from each channel.
Technical Specifications
  • Wavelength Range: 400 - 750 nm
  • Throughput: 96% (425 - 675 nm)
  • Chromatic Aberration: < +/- 0.2 mm (486 - 656 nm)
  • Distortion: < 0.5%
  • Maximum Sensor Format: 22 mm diagonal
Conclusion
Andor's imaging solutions provide flexible and versatile options for simultaneous multi-wavelength imaging, catering to a wide range of scientific applications. The devices are designed for ease of integration and use, offering high-quality imaging capabilities.
Benefits of Optosplit III
  • Cost-effective triple wavelength imaging with one camera
  • Optimized internal optics to minimize aberrations
  • Easy and quick filter set exchange
  • Customizable ROI and image size for camera sensors
  • Space-efficient design with integral C-mounts
  • Accurate image centering and pixel alignment
  • Flexibility in using multiple wavelengths
  • Acts as an adjustable neutral density filter
  • Maximizes contrast through emission polarization orientation
Principle of Operation
The optical path schematic illustrates the different optical paths of reflected and transmitted signals, which are combined at the prism and projected onto the camera.
Application and Technical Notes
Andor provides flexible solutions for multi-wavelength imaging, detailing the use of TuCam with different detectors and software. A study using Optosplit II and iXon3 EMCCD for multi-color direct STORM is highlighted.
Research Paper: Multi-color direct STORM
Lampe et al. developed SD-dSTORM, combining red emitting carbocyanine dyes with spectral demixing, requiring reduced laser power and fewer imaging frames for super-resolved reconstruction of biological nanostructures.
Technical Note: Dual Wavelength Imaging using TuCam
Guidance is provided on setting up software for use with TuCam, covering Andor Solis, Andor iQ, and MetaMorph, including software support for detectors and camera synchronization methods.
Acquisition Testing
Tests with iXon Ultra 897 EMCCD and Neo 5.5 sCMOS cameras showed frame rates consistent with single camera operation.
USB Enhanced Host Controller (EHC) Configuration
To achieve maximum data transfer rates with the iXon Ultra 897 camera, each camera should be connected to a separate USB Enhanced Host Controller (EHC). A USB hub finder tool is available to locate these ports.
Test Configuration
An in-house test successfully operated seven iXon Ultra 897 cameras simultaneously at maximum frame rate using a Dell Precision T5600.
Part Numbers and Accessories
The document lists part numbers for mounting types, magnification options, and standard filter sets, along with accessories for Optosplit II and III configurations.
Software Recommendations
For dual wavelength image acquisition using two cameras on TuCam, compatible software includes Andor Solis, Andor iQ, MetaMorph, and NIS-Elements.
Customer Support
Andor provides extensive customer support services, including on-site assistance, training, and testing services, with extended warranty packages available.
Contact Information
Contact details for Andor's offices in Northern Ireland, North America, Japan, and China are provided for customer support and inquiries.
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Catalog excerpts

Multi-Wavelength Imaging-1

ANDOR an Oxford Instruments company Multi-Wavelength Imaging Solutions For Simultaneous Capture

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Multi-Wavelength Imaging-2

Multi-Wavelength Imaging Solutions For Simultaneous Capture The vast majority of fluorescence microscopy applications involve the use of more than one fluorescent probe e.g. FRET, dual color imaging, co-localization studies etc. For simultaneous multi-wavelength imaging Andor offers the most flexible and versatile solutions available, either splitting the wavelengths across two separate cameras or across adjacent sensor halves of a single camera. Image courtesy of Dr. Ulrike Engel, Nikon Imaging Centre, University of Heidelberg, Germany The TuCam, a new generation two-camera adaptor, for both...

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Multi-Wavelength Imaging-3

an Oxford Instruments company TuCam Dual Image Adapter Andor's TuCam is a new generation two-camera adapter for macro or microscopic imaging applications. Available in C-mount, TuCam's features include large aperture, exceptional transmission, very low distortion and high precision alignment using kinematic cassettes. The TuCam can be configured for simultaneous imaging from two similar cameras or as a switch between camera models with different imaging capabilities, including CCD, EMCCD, ICCD and sCMOS from Andor, and other third party detectors. Since separate wavelengths can be split across...

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Multi-Wavelength Imaging-4

an Oxford Instruments company TuCamDual Image Adapter Principle of Operation This schematic of the optical path shows the emission beam (yellow) coming from the microscope and entering the TuCam via the C-mount attachment point. When this emission beam enters the filter cassette, which contains a dichroic mirror, the emission light is split. The longer wavelengths are transmitted straight through the dichroic mirror (red beam) and collected by camera one and the shorter wavelengths of light are reflected by the dichroic mirror (green beam) and collected by the second camera. Dichroic mirror and...

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Multi-Wavelength Imaging-5

an Oxford Instruments company Optosplit II One Camera Solution - Dual Emission Image Splitter The Optosplit II image splitter is an elegant device that divides the image into two separate, spatially equivalent components that can be displayed side by side on a camera sensor, enabling a single camera to record images simultaneously at two different optical wavelengths. The Optosplit has been designed as a convenient, inexpensive solution to simultaneous imaging. Splitting is usually performed on a basis of wavelength, allowing applications such as ratiometric ion imaging or FRET, however, polarizing...

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Multi-Wavelength Imaging-6

an Oxford Instruments company Optosplit IIIOne Camera Solution - Triple Emission Image Splitter The Optosplit III, a three-way image splitter, is a simple device for dividing an image into two or three separate, spatially equivalent components. These can be displayed side by side on a camera sensor, enabling a single camera to record images simultaneously at three different optical wavelengths. The Optosplit III has been designed as a convenient, inexpensive solution for simultaneous imaging. Splitting is usually performed on the basis of wavelength or polarization, allowing applications where...

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Multi-Wavelength Imaging-7

Application and Technical Notes For multi-wavelength imaging, Andor has the most flexible and versatile solutions available to the researcher. The following section is dedicated to providing a greater depth of understanding to the functionality of the multi-wavelength imaging products available from Andor. The content in the subsequent pages will illustrate how the TuCam can be used with different detectors in a range of software, making it easy to integrate into any laboratory. In addition, you will read a summarized publication where the Optosplit II, in combination with the iXon3 EMCCD from...

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Multi-Wavelength Imaging-8

Application Note Multi-color direct STORM with red emitting carbocyanines Lampe et al have developed a novel variant of direct STORM (dSTORM) termed spectral demixing dSTORM (SD-dSTORM) that combines the photochemical advantages of the red emitting carbocyanine dyes with the principal of spectral demixing. Specifically, they use a novel combination of carbocyanine dyes for super resolution, Alexa Fluor 647 and Alexa Fluor 700, which both show the excellent buffer compatible blinking characteristics needed for single molecule localization. SD-dSTORM requires reduced laser power and fewer imaging...

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Multi-Wavelength Imaging-9

an xford Instrument company Technical Note Dual Wavelength Imaging using TuCam: Software Set-up Focus This technical note is provided to aid the user in the set-up of their software for use with TuCam. The example softwares covered are Andor Solis, Andor iQ and MetaMorph from Molecular Devices Corporation. Table 1 - Frame rates measured from two iXon Ultra 897 EMCCD cameras operating in Solis (Version 4.21.300006) software. Table 2 - Frame rates measured from two iXon Ultra 897 EMCCD cameras operating in iQ (version 2.6) software. Table 3 - Frame rates measured from two iXon Ultra 897 EMCCD cameras...

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Multi-Wavelength Imaging-10

an xford Instrument company Technical Note Software recommendations for acquisition and analysis of dual wavelength microscopy images Recommended software packages for dual wavelength image acquisition when using two cameras on TuCam When acquiring two color images with two cameras on TuCam it is essential that the acquisition software is compatible with the detectors used and is capable of acquiring simultaneous image capture from both cameras. This section provides information concerning compatibility of Andor’s range of imaging cameras for microscopy with a number of acquisition softwares,...

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Multi-Wavelength Imaging-11

Customer Support Andor products are regularly used in critical applications and we can provide a variety of customer support services to maximize the return on your investment and ensure that your product continues to operate at its optimum performance. Andor has customer support teams located across North America, Asia and Europe, allowing us to provide local technical assistance and advice. Requests for support can be made at any time by contacting our technical support team at andor.com/support. Andor offers a variety of support under the following format: • On-site product specialists can...

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