A NOVEL AND EFFICIENT METHOD FOR DIGITAL MICROSCOPY OF CELLS IN CULTURE

Abstract

Common implementations of standard microscopy suffer from several disadvantages. The apparatus required is often inconveniently large and expensive, requires trained operators, provides only a limited field of view (FOV) at high magnification, and requires continuous manual refocusing or expensive automated corrective focusing attachments. Furthermore, long-term imaging of living cells or tissues generally requires expensive microscope incubators, as well as automated, motorized repositioning and tiling systems to achieve a large field of view. In our laboratory, we have developed a form of lensless microscopy using enhanced CMOS image sensors. This implementation of lensless microscopy, namely contact optical microscopy (COM), provides unprecedented convenience and low cost. This approach provides sub-micron resolution across more than 100-fold larger fields of view, continuously in focus without the requirement for repositioning or tiling. Additionally, the small form factor (handheld and portable) of these new instruments permits within-incubator imaging. In this research, we have developed the lensless imaging platform and proven that even primary dissociated cells, normally requiring the most fastidious culturing conditions, can be grown in culture on such devices for extended periods permitting long-term time-lapse video microscopy, and that cells in suspension can be immunolabelled and enumerated with immunocytometry accuracy essentially equivalent to gold-standard fluorescence flow cytometers.