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Leica DMIRB Inverted Fluorescence Microscope

Introduction

The Leica DMIRB is a computer-controlled conventional wide-angle (non-confocal) microscope. Its inverted nature means it is uniquely designed for use with cell culture experiments. It is equipped with a stage accepting a range of plates and microscope slides and a high-resolution CCD camera. There is maximum flexibility in using the microscope for effective contrasting methods such as Brightfield, Phase Contrast, Darkfield Contrast, Polarization Contrast, Leica Modulation Contrast, DIC and Fluorescence. The microscope is fixed with a Solent Scientific transparent environmental chamber. This incubation chamber is used for prolonged studies of living cells, including time lapse image capture experiments for cell motility and cell death. The Leica DMIRB is a viable alternative to confocal microscopy that is not always the best solution for imaging needs and, in some situations, may even be counter-productive. The Leica DMIRB can produce similar results and offers as much flexibility. The dedicated FW4000TZ software handles image captures and use and offers a total fluorescence imaging solution.

Leica DMIRB

Technical Specifications

'Live Cell' Option

Microscopy – Leica Modulation Contrast optics. 10, 20 and 40x objectives.

Environment – Solent Scientific transparent environmental chamber. Warm, filtered air circulates within the acrylic chamber from a heater unit that is mechanically isolated from the environmental chamber. Two doors allow specimens to be changed and the condenser settings to be adjusted. A further two doors, below the stage, allow access to the nosepiece and objectives. Focusing and stage controls remain outside the environmental chamber. The 35mm camera, video camera, multi-viewing and fluorescence capabilities of the microscope are unaffected. The chamber can, when required, be quickly removed without the use of hand tools.

Microscope Control – FW4000TZ software (see below) allows user to perform time-lapse experiments with automated shutter control to prevent excessive exposure of specimen to transmitted light. The motorised focus allows for Z-stacking (multiple focus during experiments).

Post experiment processing – Movies generated by the FW4000 software can be converted and analysed with DIAS Software. This system represents the most advanced computer system commercially available for analyzing how cells move and change shape overtime. It contains both manual and automatic digitization modes, advanced image-processing capabilities, the ability to quantitate more than 30  parameters of motion and dynamic morphology, the capacity to generate a variety of movies demonstrating motility and dynamic morphology, and sophisticated graphing and analytical capabilities.

'Fixed Cell' Option

Microscopy - Brightfield, Phase Contrast, Darkfield Contrast, Polarization Contrast and Fluorescence. 20, 40 and 63x objective. Optimized Fluorescence with a 4 position filter cube turret (manual) without pixel shift, and filters for UV (dapi), FITC, cy3 and cy5

Basic Acquisition Suite

Acquisition
Basic Enhancement Tools (Contrast, brightness, gamma)
Lab Book
Basic Printing
Annotation
Annotation & Calibration
Probe meter and pseudo-colouring
Zoom and pan
Composite creation
Channel mixing
Z-stack acquisition
Time sequence builder
lapse experiments Compile web documents
Auto Relocate

Enhancements Module

Region of interest
No neighbours deconvolution
Sharpening/smoothing tools
Background removal
Nearest neighbours deconvolution
Maximum intensity projections

Measurement Module

Manual measurements
Automatic measurements
Object counting
Grey level profiling

Deblur And 3D Visualize Module
Gallery Module

Interactively review experiments
Create composites on collections of image
Filter and display selected images
Select images for processing

Publish Module

Compiling. avi movies
Printing reports
Print 1,2 or 4 images
Printing image montages of Z-stacks and time

Examples Of Work Undertaken At The Institute

Examples

Examples in “fixed cell” mode

LEFT: Human jaw muscle cells fluorescently stained for cytoskeletal markers (red and green) and DNA (blue).
RIGHT: Human jaw muscle cells identified by phase contrast (greyscale) and DNA (blue).

Examples in “live cell” mode

Human jaw muscle cells were cultured on soluble glass fibres band monitored with live time lapse.


For more information, please contact
Professor Jonathan Knowles
Email. j.knowles@ucl.ac.uk
Tel. +44 (0)20 3456 1189
Eastman Dental Institute
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