An Alternative to Dye-Based Approaches to Remove Background Autofluorescence From Primate Brain Tissue

Brain tissue contains autofluorescing elements that potentially impede accurate identification of neurons when visualized with fluorescent microscopy. Age-related accumulation of molecules with autofluorescent properties, such as lipofuscin, can possess spectral profiles that invade the typical emission range of fluorophores commonly utilized in fluorescent microscopy. The traditional method for accounting for this native fluorescence is to apply lipophilic dyes that are able to sequester these unwanted signals. While effective, such dyes can present a range of problems including the obstruction of fluorescent probe emissions. The present study utilizes aged primate midbrain tissue stained for tyrosine hydroxylase and calbindin to investigate an image processing approach for removing autofluorescence utilizing spectral imaging and linear unmixing. This technique is then compared against the traditional, dye-based autofluorescence sequestration method using Sudan Black B (SBB). Spectral imaging and linear unmixing yielded significantly higher cell numbers than SBB treatment. This finding suggests that computational approaches for removing autofluorescence in neural tissue are both viable and preferential to dye-based approaches for estimation of cell body numbers.NIA [R01 AG050548, P51 RR000169, F31 AG055263]; McKnight Brain Research FoundationOpen access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Fabrication of metal nanoparticle arrays by controlled decomposition of polymer particles

We report a novel fabrication method for ordered arrays of metal nanoparticles that exploits the uniform arrangement of polymer beads deposited as close-packed monolayers. In contrast to colloidal lithography that applies particles as masks, we used thermal decomposition of the metal-covered particles to precisely define metal structures. Large arrays of noble metal (Au, Ag, Pt) nanoparticles were produced in a three-step process on silicon, fused silica and sapphire substrates, demonstrating the generality of this approach. Polystyrene spheres with diameters ranging between 110 nm and 1 μm were convectively assembled into crystalline monolayers, coated with metal and annealed in a resistive furnace or using an ethanol flame. The thermal decomposition of the polymer microspheres converted the metal layer into particles arranged in hexagonal arrays that preserved the order of the original monolayer. Both the particle size and the interparticle distance were adjusted via the thickness of the metal coating and the sphere diameter, respectively.

Do individual-tree growth models correctly represent height:diameter ratios of Norway spruce and Scots pine?

▶ We examined four individual tree models in Europe: BWIN, Moses, Silva and Prognaus. ▶ We simulated growth of open-grown trees and on research plots for 15 or 30 years. ▶ Height:diameter ratios were correctly predicted by all four models. ▶ Height:diameter ratios were within the bounds of open grown trees and dense stands. ▶ They decreased with age and density; dominant trees had lower ratios than mean trees