Subcellular Microanatomy by 3D Deconvolution Brightfield Microscopy: Method and Analysis Using Human Chromatin in the Interphase Nucleus

Figure 1

Partial volume of tissue including a whole hepatocyte nucleus before (left) and after (right) deconvolution. The volume in both cases is shown from two different angles (top and bottom). In the predeconvolved volume the smearing of structural details in the 𝑍-direction prevents detailed structural analysis. The brightfield deconvolution method developed for this work greatly reduces this smearing. However, the deconvolved cytoplasmic details partly obscure the view of the nuclear chromatin within. As the aim of the present paper is to make studies of the nuclear chromatin, the extraneous information of the cytoplasmic structure is manually dissected away from the digital images as part of the image postprocessing steps described in the text.