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P8005
Label-free Analysis of Tissue Sections & Cells Using Laser Scanning Reflectance Imaging
Presenter Wayne Bowen, TTP LabTech Ltd, United Kingdom
Additional Authors: Tristan Cope, David Billington
Brightfield transmission microscopy is the most common method for viewing tissue sections and cells in scientific and clinical diagnostic laboratories. Simple staining procedures and ease of use make it a readily accessible technology, however, there are drawbacks for some applications. For example, when examining large tissue sections the limited field of view afforded by microscope objective lenses can make analysis of large areas a laborious and prolonged procedure. In addition, when automated microscopes are employed individual images need to be stitched together. The small field of view also limits the number of cells that can be examined when they are cultured in microplates for cell-based screening assays.Laser-scanning microplate cytometers, such as TTP LabTech’s Acumen® eX3, represent a possible alternative platform for certain applications. Such technology is routinely used for the high-content screening of fluorescent cells since it offers rapid analysis over a large field of view (20 mm x 20 mm – approximately 400 times greater than a X10 microscope objective). The Acumen eX3 is capable of generating TIFF images (8 or 16-bit) of the raw data received from the sample which can be subjected to the full range of algorithms contained in commercially available image analysis software. In this study, modifications were made to the collection optics of an Acumen eX3 so that light reflected by the sample could be detected as the laser spot rastered across the sample. This technique called Laser Scanning Reflectance Imaging (LSRI) was used successfully to scan a range of unstained tissue samples mounted on glass microscope slides using a 488nm laser. The resultant images correlated well with observations made using traditional phase contrast brightfield microscopy. In microplates, LRSI required the removal of culture medium to increase the contrast of cells.In summary, LSRI using microplate cytometers offers rapid label-free imaging of tissue sections and cells in an automated workflow for more efficient analysis, and the potential for multiplexing with fluorescent staining protocols.