Embedding complementary imaging data in laser scanning microscopy micrographs by reversible watermarking.

Ioan-Catalin Dragoi,Radu Hristu,Marius Popescu,Henri-George Coanda,Denis E Tranca,Stefan G Stanciu,Dinu Coltuc

doi:10.1364/boe.7.001127

Ioan-Catalin Dragoi, Radu Hristu + Show 5 more

Open Access

https://doi.org/10.1364/boe.7.001127

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Abstract

Complementary laser scanning microscopy micrographs are considered as pairs consisting in a master image (MI) and a slave image (SI), the latter with potential for facilitating the interpretation of the MI. We propose a strategy based on reversible watermarking for embedding a lossy compressed version of the SI into the MI. The use of reversible watermarking ensures the exact recovery of the host image. By storing and/or transmitting the watermarked MI in a single file, the information contained in both images that constitute the pair is made available to a potential end-user, which simplifies data association and transfer. Examples are presented using support images collected by two complementary techniques, confocal scanning laser microscopy and transmission laser scanning microscopy, on Hematoxylin and Eosin stained tissue fragments. A strategy for minimizing the watermarking distortions of the MI, while preserving the content of the SI, is discussed in detail.

Highlights

Laser Scanning Microscopy (LSM) techniques represent essential tools for multiple fields of science
While Confocal Laser Scanning Microscopy (CLSM) can be used to acquire optical sections at specific depths based on the reflected light or on the fluorescence emission, Transmission Laser Scanning Microscopy (TLSM) provides information on how the excitation light is transmitted through the whole specimen volume, and on how the excitation light is absorbed in this volume
We present the results of embedding by local prediction reversible watermarking the lossy compressed TLSM images of Fig. 2 into the corresponding CLSM ones and vice versa

Summary

Introduction

Laser Scanning Microscopy (LSM) techniques represent essential tools for multiple fields of science. For example Second Harmonic Generation Microscopy (SHG) is a perfect choice for imaging noncentrosymetric structures, no matter if these are fluorescent or not, but cannot be used to image isotropic structures, whereas Two-Photon Excitation Fluorescence Microscopy (TPEF) can image fluorescent structures, irrespectively of their isotropy, but cannot be used to image non-fluorescent ones [2, 4, 5] For this reason, in order to acquire an in-depth understanding of an investigated sample, LSM users frequently need to image it using different LSM techniques and workmodes, under different acquisition configurations or at different moments in time. The resulting images typically need to be stored, retrieved and transmitted, together, so that the data sets can be associated, analyzed and interpreted at a later date or by different users

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