Subsamples of between 1 and 3 cm3 were prepared for pollen analysis using standard HCl, sieving, HF, acetolysis and density separation techniques (solution density 1.9) outlined in Moore et al. [30 ]. Lycopodium tablets [31 ] were added to allow the calculation of pollen and microscopic charcoal concentration and influx data. After processing, the samples were suspended in silicone oil prior to being mounted on slides. Counting was undertaken using a Leica DM 2500M light microscope at x500 magnification (with x1000 used for critical observations) until a sum of 500 total land pollen (TLP) had been achieved. Pollen and spores were identified with the aid of the key in Moore et al. [30 ], photographs in Reille [32 ] and modern reference material, with nomenclature following Beug [33 ]. Cereal-type pollen grains were categorised following measurements in Andersen [34 ] and non-pollen palynomorphs (NPPs) were identified using van Geel et al. [35 ] and van Geel and Aproot [36 ]. Pollen diagrams were constructed using TILIA and TGView software [37 ] and percentages were based upon the TLP sum minus Lactuceae (see below for justification).
Dm 2500m light microscope
Manufactured by Leica
The Leica DM 2500 M is a light microscope designed for routine microscopy tasks. It features a stable, ergonomic stand and provides high-quality optics for clear, detailed observations. The microscope is equipped with various magnification options and can be used for a range of applications in research and industrial settings.
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3 protocols using dm 2500m light microscope
1
Pollen Analysis of Sediment Samples
2
Multimodal Imaging Workflow for Xenograft Analysis
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Morphological Analysis for Quantification Morphometry and measurements were performed with Fiji/ImageJ software. For quantifications, at least 4-5 different brain sections comprising the human xenografts and the adjacent mouse host tissue were included per animal.
Immunofluorescence (IF) sections were imaged by confocal microscopy (Nikon Ti-E inverted microscope) with a 20x (0.75 NA) objective to image Z series stacks of areas inside human xenografts or mouse host tissue (8-10 optical sections 1 mm apart for each image). All images were acquired using identical acquisition parameters as 16-bit, 1024x1024 arrays. Z series stacks were then converted in Fiji/ImageJ to maximum intensity projections and threshold adjusted to isolate specific fluorescence.
Immunohistochemistry (IHC) sections were imaged using a Leica DM 2500M light microscope with a 63x objective. All images were acquired using identical acquisition parameters as 16-bit, 1024x1024 arrays.
Ultrathin EM sections comprising either the human xenografts or the adjacent mouse host tissue were imaged using a JEM-1400 transmission electron microscope (JEOL) equipped with a 11M-pixel Olympus SIS Quemesa camera. Images were taken in randomly selected areas at a 600x magnification.
Immunofluorescence (IF) sections were imaged by confocal microscopy (Nikon Ti-E inverted microscope) with a 20x (0.75 NA) objective to image Z series stacks of areas inside human xenografts or mouse host tissue (8-10 optical sections 1 mm apart for each image). All images were acquired using identical acquisition parameters as 16-bit, 1024x1024 arrays. Z series stacks were then converted in Fiji/ImageJ to maximum intensity projections and threshold adjusted to isolate specific fluorescence.
Immunohistochemistry (IHC) sections were imaged using a Leica DM 2500M light microscope with a 63x objective. All images were acquired using identical acquisition parameters as 16-bit, 1024x1024 arrays.
Ultrathin EM sections comprising either the human xenografts or the adjacent mouse host tissue were imaged using a JEM-1400 transmission electron microscope (JEOL) equipped with a 11M-pixel Olympus SIS Quemesa camera. Images were taken in randomly selected areas at a 600x magnification.
3
Emulsions with Silica Particles
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Emulsions are first prepared by mixing 3% w/w monodisperse 1.5 μm diameter precipitated silica particles (Nippon-Shokubai KE-P150) into hexadecane (Sigma-Aldrich, 99%) [9] .
A volume of the silicahexadecane dispersion is then emulsified into an equal volume of deionized water by manual shaking for three minutes. The emulsion was then aged for 24 hours and inspection revealed a small fraction of elongated droplets. Imaging of the droplets is carried out on a Leica DM2500M light microscope using phase contrast optics.
A volume of the silicahexadecane dispersion is then emulsified into an equal volume of deionized water by manual shaking for three minutes. The emulsion was then aged for 24 hours and inspection revealed a small fraction of elongated droplets. Imaging of the droplets is carried out on a Leica DM2500M light microscope using phase contrast optics.
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