These are: (1) the use of an array of UV-LEDs to increase the amount of UV energy that is applied to the polymer coated glass slides, (2) the fabrication of a curing platform with two such UV-LED arrays, which allows for the convenient collection of alternating sections for parallel processing in two slide series, (3) determining optimal parameters for tissue preparation and sectioning temperatures. We have made several hardware and procedural modifications to the CryoJane product that resulted in a significantly enhanced sectioning system. After polymerization, the section binds with the slide and the overlying tape can easily be peeled off. The tape and section together are placed onto a polymer-coated glass slide that polymerizes under ultraviolet (UV) light. In this procedure, adhesive tape is attached to the blockface, the block is sectioned without the standard anti-roll plate, and the detached, cut section remains adhered to the tape without curling or other deformation. Scale bars are 1mm.Ī commercially available tape system (CryoJane Instrumedics/Leica Microsystems) has been developed to facilitate the transfer of cut sections from the blockface to the slide. Allen Reference Atlas, coronal Level 73 is shown in (A) level 83 is shown in (B) level 84 is shown in (C) and level 96 is shown in (D). The three most common sources of damage are: folded areas (solid black arrows), as in A, B and C torn sections (arrowheads, as in C), and sections with missing areas (dotted arrows, in D). A scan through the Allen Reference Atlas, one of the most commonly used anatomical references of the mouse, turns up examples of section damage (arrows). Obtaining undamaged cryostat sectioned material is difficult, especially for relatively thin sections (20–25μm). Common examples of section damage (downloaded from the Allen Reference Atlas). The second method has the advantage of minimizing the degree of section handling, but will result in torn, wrinkled, or curled sections unless the temperature differential between the sections and pick-up slide is optimal.įig 1. Two basic such methods are: (1) using a fine brush to transfer sections to solution for further, free-floating processing or for mounting on slides, (2) directly pressing sections onto glass slides from the anti-roll plate of the cryostat. In conventional cryostat sectioning, there are many methods of collecting individual sections as these are cut (“shaved”) off the main block of tissue. It is, however, labor-intensive and even for expert users, there is some risk of lost or damaged sections ( Fig 1). In this report, we describe a protocol for tape-assisted cryosectioning which significantly improves section quality and the speed of collecting brain sections.Ĭryostat tissue sectioning is a popular means of sectioning brain tissue for further processing, imaging, and analysis. Ease of processing is another factor, especially in the very common case where processing is being carried out by students or non-expert technical staff. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist.Ī basic component of the neuroanatomical workflow is efficient and reliable histological processing and subsequent analysis and correct interpretation of experimental results heavily depend on high-quality histological sections. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedįunding: Support was provided by the Challenge Grant from the National Institutes of Health (RC1MH088659), PPM Transformative Award from the Office of the NIH Director (R01MH087988), PPM. Received: ApAccepted: Published: July 16, 2015Ĭopyright: © 2015 Pinskiy et al. Borchelt, University of Florida, UNITED STATES Citation: Pinskiy V, Jones J, Tolpygo AS, Franciotti N, Weber K, Mitra PP (2015) High-Throughput Method of Whole-Brain Sectioning, Using the Tape-Transfer Technique.
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