Position-referenced microscopy for live cell culture monitoring

Position-referenced microscopy (PRM) is based on smart sample holders that integrate a position reference pattern (PRP) in their depth, allowing the determination of the lateral coordinates with respect to the sample-holder itself. Regions of interest can thus be retrieved easily after culture dish transfers from a cell incubator to the microscope stage. Images recorded at different instants in time are superimposed in a common coordinate system with subpixel accuracy. This paper presents such smart Petri culture dishes and their use for live cell culture monitoring. The impact of the PRP on the light budget is discussed and performances are demonstrated. First results on the application of PRM to the observation of apoptotic body internalization are reported.

[1]  J. Goodman Introduction to Fourier optics , 1969 .

[2]  P. Toner,et al.  An improved method for sequential light and scanning electron microscopy of the same cell using localising microcoverslips. , 1982, Journal of clinical pathology.

[3]  Patrick Sandoz,et al.  Pseudo-Periodic Encryption of Extended 2-D Surfaces for High Accurate Recovery of any Random Zone by Vision , 2010 .

[4]  Harald zur Hausen,et al.  Papillomaviruses in the causation of human cancers - a brief historical account. , 2009, Virology.

[5]  S. Orrenius,et al.  Apoptosis: a basic biological phenomenon with wide‐ranging implications in human disease , 2005, Journal of internal medicine.

[6]  F. Wyrowski,et al.  Fast calculation method for optical diffraction on tilted planes by use of the angular spectrum of plane waves. , 2003, Journal of the Optical Society of America. A, Optics, image science, and vision.

[7]  Laurent Robert,et al.  Position encryption of extended surfaces for subpixel localization of small-sized fields of observation , 2009, 2009 International Symposium on Optomechatronic Technologies.

[8]  F. Ruddle,et al.  Photoengraving of coverslips and slides to facilitate monitoring of micromanipulated cells or chromosome spreads. , 1981, Experimental cell research.

[9]  Fluorescence microscopy of living cells in culture. Part B. Quantitative fluorescence microscopy--imaging and spectroscopy. , 1989, Methods in cell biology.

[10]  R. Webb,et al.  In vivo confocal scanning laser microscopy of human skin: melanin provides strong contrast. , 1995, The Journal of investigative dermatology.

[11]  D. St-Jacques,et al.  Nanoscale grid based positioning system for miniature instrumented robots , 2003, CCECE 2003 - Canadian Conference on Electrical and Computer Engineering. Toward a Caring and Humane Technology (Cat. No.03CH37436).

[12]  Robert M. Clegg,et al.  Fluorescence lifetime imaging microscopy (FLIM): Spatial resolution of microstructures on the nanosecond time scale , 1993 .

[13]  J. Fujimoto Optical coherence tomography for ultrahigh resolution in vivo imaging , 2003, Nature Biotechnology.

[14]  John G. Webster,et al.  The Measurement, Instrumentation and Sensors Handbook , 1998 .

[15]  P Sandoz,et al.  Position referencing in optical microscopy thanks to sample holders with out‐of‐focus encoded patterns , 2007, Journal of microscopy.

[16]  P. Marquet,et al.  On the complex three‐dimensional amplitude point spread function of lenses and microscope objectives: theoretical aspects, simulations and measurements by digital holography , 2007, Journal of microscopy.