We offer an advanced montaging system, which we call the Tiled Field Mapping system. TFM is available as part of the MCID Elite or AIS image analyzers. It offers a unique feature set.
Integration with a powerful image analyzer: TFM is an integral part of our state-of-the-art; multi-functional MCID™ Elite image analysis system, and is also available as an application-specific AIS™ Montaging system. Both systems provide a natural and convenient interface for working with large images, including image enhancement, archiving, printing, manipulation and quantification.
Autoalignment: Spatial errors accumulate as a motor stage moves over larger distances. The edges of two adjacent fields might be within a pixel, but the error will be multiple pixels three fields away. TFM includes algorithms that perform automated alignment of the discrete fields, and feedback regulation of stage precision as the montaging proceeds.
Intensity/color correction: Cameras, microscope optics and illumination systems all contribute to intensity variations (e.g. 20%) across a field of view. There are also color variations that might not be obvious in a single field, but any intensity and/or color variation that repeats from tile to tile will form a pattern that is very obvious in a montage. Therefore, TFM corrects both intensity and color errors, automatically.
Autofocus: Residual tilt in the microscope stage is not noticed within a single field, but is very obvious across multiple fields of view. Point-to-point variations in specimen depth a present both within and across fields. TFM includes automatic corrections and accommodations which achieve sharply focused images.
- A tilt correction function adjusts for stage misalignment.
- Software autofocus makes a number of exposures and calculates the best plane of focus for each tile.
- A hardware autofocus system (laser detector, stage with fast Z positioning) is an available option. It operates in conjunction with the software autofocus and speeds the focus process.
- Image combination convolves any number of planes of focus into a single sharp image. The image combination feature is particularly useful for thick specimens.
Any size image: Unlike many other montaging systems, TFM does not limit the number of tiles that can be acquired. Start it up and leave it running. As a general rule, expect about 10 seconds/tile. We have routinely made montages of about 8000 x 8000 pixels, containing more than 200 microscope fields.
Fluorescence montages: TFM uses any of the monochrome or color cameras that the MCID/AIS image analyzers support. These include cameras suitable for low light imaging of fluorescent or luminescent samples (Fig. 6).
Any microscope: TFM can be used with any microscope, upright or inverted using one of many leading motorised stage kits from Prior, Ludl & Marzhauser. Visit our stage section for more detailed information on the options available.
Fluorescence montage consisting of 28microscope fields (10X objective, monochrome camera, three color excitation presented with a computer-controlled filter wheel) acquired with an MCID Elite system. The image is a xenograft of a human cervical cancer cell line grown in mouse. The red areas are regions of hypoxia, the green are blood vessels and the blue is Hoechst- labeled DNA.