The new Leica Application Suite Advanced Fluorescence LAS AF 4.0 combines the most powerful features available today in microscope software in one package, while focusing on usability in all aspects of interface, function and workflow.
Obtain reproducible analysis results rapidly and easily with the 2D image analysis wizard. Go step by step through the guided workflow from applying filters, thresholding, binary image processing through measurements and classification. Use a binary reference mask in multi-channel image analysis, for example to count the number of spots in each nucleus.
Save the analysis results with the experiment, export the results to Excel or create a report including histogram and binary masks.
For repetitive analysis tasks, user-defined protocols can be saved and applied to new data sets. Several data sets can be added to a batch list and processed at once. Results from different images can also be accumulated, providing statistical information.
Create reproducible results step by step using the 2D Analysis workflow. Each step offers a set of analysis tools. The viewer provides immediate feedback on the applied settings. Use automatic tools such as auto threshold, split objects, or edge removal to run through an analysis automatically. Set pauses to allow user interaction during automated analysis, for example for binary image editing.
Interactively select objects in the viewer and highlight measurement results in the results table or vice versa. Select from a large number of measurement parameters. Rearrange the order of measurement columns and use columns to sort results. Export measurements and statistics to Excel or save them with the experiment. Reproduce measurements easily with user defined protocols.
Use different analysis channels for different fluorescence stains and analyze all aspects of your sample at the same time. An individual workflow can be assigned to each analysis channel.
Apply a binary reference mask to obtain object specific data, for example count the number of spots in each nucleus.
Alternatively, apply multiple analysis channels to the same image. On a stained color image you can for example count the healthy cells in analysis channel 1 and use a second analysis channel to count the abnormal cells.
Get unbiased and reproducible classification results even in situations where thresholding or filtering is not suitable. Group your results with the intelligent classifier and show them in a histogram. Train the classifier on a training data set. Save classifiers and apply to new data sets. A support vector machine divides the new objects into the predefined classes.
Have full control of your experimental conditions with the LAS AF Environmental Control module. Log the environmental data and monitor it during the experiment. Set all environmental conditions within one interface and run temperature profiles, for example for heat shock experiments.
All environmental data is saved with the experiment. Review the environmental conditions subsequent to an experiment. Exclude temperature, CO2 or O2 conditions from being the cause of a failed experiment.
In combination with LAS AF Live Data Mode macros can be generated to run temperature profiles, for example for heat shock experiments. Acquisition can be started once a predefined temperature is reached. During waiting time a message is displayed showing the difference between nominal and actual environmental parameters. Alternatively the acquisition can commence during ramp up of temperature, for example.
All environmental parameters are set within one interface. Activate range control to get a warning when upper or lower limits are exceeded. Alternatively the experiment can be stopped automatically once a limit is passed.
Don´t miss anything! With the LAS AF Mobile Connection module you can stay connected to your experiment anywhere at any time. Review your experiment or download a specific image in full resolution and send it to a colleague. Start, pause, or stop experiments, or capture an image to identify the best time for starting your experiment remotely. The imaging conditions are displayed on your remote device.
Connect to your acquisition station via web client or mobile device and see the course of your experiment, review the whole experiment or open any image series available in the experiment tree.
Review the channel settings of your experiment such as exposure time, gain, and filter cube or take a look at the environmental conditions such as temperature, CO2 or O2 concentration (requires LAS AF Environmental Control module). The experiment settings are defined in advance on the acquisition station.
Multiple “Remote Users” can be connected with the system at the same time. Chat with connected colleagues and send emails to other colleagues. Send messages to the user at the acquisition station.
Courtesy of Lionel Newton, EMBL Heidelberg
Green: Lateral line primordium (GFP)
Red: Neuronal marker (DsRed)
Blue: Nucleus marker (BFP)
Gray: Muscles (SHG)
Courtesy of Darren Gilmour, EMBL Heidelberg
Green: Mechanosensory lateral line placode (GFP)
Red: Innervating axons (DsRedExpress)
Courtesy of Yuya Ohhara, Satoru Kobayashi, National Institute for Basic Biology, Japan
Green: Neuroendocrine cells (GFP)
Violet: Actin filaments (Phalloidin)
To extend the range of applications for both widefield and confocal systems, optional packages are offered for Colocalization Analysis, Calcium Imaging, Electrophysiology, Triggering and complex timelapse experiments, FRET, photobleaching, 3D Visualization with real time rendering, Deconvolution.
All application, processing and quantification modules are identical for all platforms and are fully integrated into LAS AF for fast handling and visualization of large data sets.
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