Journal: BMC Medical Imaging

Article Title: Extended Field Laser Confocal Microscopy (EFLCM): Combining automated Gigapixel image capture with in silico virtual microscopy

doi: 10.1186/1471-2342-8-13

Figure Lengend Snippet: Breaking the Gigapixel limit of a confluent confocal panorama image : A 2,8 Gigapixel size confocal mosaic, built up from 3000 (50 columns × 60 rows) individual confocal images (A), using the ImageLab/Virtual Microscope software. The internal structures of a mouse head are visualized with eosin fluorescence at 63X/NA1.25, individual images cover an area of 135 μm × 103 μm (B), captured at binning 1 (1344 × 1024 pixels) and 200 pixels overlap. After image processing in the ImageLab/Virtual Microscope software the final seamless confocal panorama show an area of 5,75 mm × 4,98 mm. The original resolution of each individual image is preserved in the final montage, allowing the user to zoom in on details at single cell level (C).

Article Snippet: The system includes: A motorized Axiovert fluorescence microscope 200 M (Zeiss GmBH, Göttingen, Germany), a CSU10 Yokogawa head – a multilens/multi pinhole array aperture (Yokogawa, Japan), a motorized XY-table (Märzhauser, Germany), an ORCA ER cold CCD camera; detector array 1344 × 1024 px (Hammamatsu, Japan), a 3-line Argon-Krypton Laser; 488 nm, 568 nm, 647 nm (Melles Griot, USA) and a mercury lamp (Osram) for 365 nm illumination.

Techniques: Microscopy, Software, Fluorescence

Journal: BMC Medical Imaging

Article Title: Extended Field Laser Confocal Microscopy (EFLCM): Combining automated Gigapixel image capture with in silico virtual microscopy

doi: 10.1186/1471-2342-8-13

Figure Lengend Snippet: Example of application of the EFLCM method : To visualize thousands of cells growing in monolayer – for automatic and objective quantitation of epigenetic effects of the transformation associated viral nuclear antigen HHV8 LANA fused to green fluorescence protein in a transient transfection assay. The blue color is Hoecsht 33342 fluorescence showing DNA. The mosaic is built up from 100 images (10 columns × 10 rows) captured at 16X/NA O.5. Individual images covers an area of 522 μm × 398 μm. Images are captured at binning 1 (1344 × 1024 pixels) with 100 pixels overlap resulting in a final area of the seamless panorama, after image processing in the ImageLab/Virtual Microscope, of 4,82 mm × 3,58 mm.

Article Snippet: The system includes: A motorized Axiovert fluorescence microscope 200 M (Zeiss GmBH, Göttingen, Germany), a CSU10 Yokogawa head – a multilens/multi pinhole array aperture (Yokogawa, Japan), a motorized XY-table (Märzhauser, Germany), an ORCA ER cold CCD camera; detector array 1344 × 1024 px (Hammamatsu, Japan), a 3-line Argon-Krypton Laser; 488 nm, 568 nm, 647 nm (Melles Griot, USA) and a mercury lamp (Osram) for 365 nm illumination.

Techniques: Quantitation Assay, Transformation Assay, Fluorescence, Transient Transfection Assay, Microscopy