Laser autofocus and image-based (or software-based) autofocus are two methods of autofocus widely used in microscopy. The Agilent BioTek patented laser autofocus method for Agilent BioTek Lionheart automated microscopes and Agilent BioTek Cytation cell imaging readers offers all the key advantages of laser autofocus:
- Speed: The laser is flashed once per well, faster than the multiple images taken during image-based autofocus. Laser autofocus works well with dim fluorophores and transmitted light imaging.
- Reproducibility: Laser autofocus is less likely to be impacted by debris in the well, since the focus is relative to the well bottom. Laser autofocus offers the highest accuracy during long-term kinetics
- Lower phototoxicity and photobleaching: Because the laser is flashed only once per well and is a very focused light source, phototoxicity and photobleaching are kept to a minimum. Laser autofocus is ideal for live cell assays and fluorophores that bleach easily.
With both image-based and laser autofocus available in Lionheart FX and Cytation, an added advantage is that users can select the most appropriate autofocus methods for the biology being imaged. The autofocus capabilities can be applied to a cell monolayer or cells moving in the well.
The laser autofocus design incorporates all components into a single cube (shown right), designed for easy user installation into the LED/filter cube slider, on new or existing instruments. Three fluorescence and one brightfield channel are available for imaging, even with the laser autofocus option installed.
Figure 1. Agilent BioTek method of laser autofocus uses a laser and the camera to locate the well bottom.
Laser Autofocus Benefits:
- Increased focus speed
- Increased reproducibility
- Lowered phototoxicity and photobleaching
- Used along with standard image-based autofocus, both methods provide best autofocus options for a variety of biologies.
This Technology is Available in:
- Agilent BioTek Lionheart FX automated microscope
- Agilent BioTek Lionheart LX automated microscope
- Agilent BioTek Cytation 5 cell imaging multimode reader
- Agilent BioTek Cytation 1 cell imaging multimode reader