Images were collected using a Nikon A1R Eclipse Ti microscope and 40x or 60x objectives. We measured the size of fat body cells when the optical section bisected the nucleus, by outlining the cell boundary (as defined by phalloidin) with the freehand selection tool. We measured LD density within each cell by determining the percentage of pixels that had LD‐associated fluorescence. LD size was measured using an optimized macro that detected single LD particles (analyze particles size = 5‐infinity) after smoothening and applying a threshold that was manually verified to specifically detect LD. Nuclear size was measured from images of anti‐LaminDm0‐labeled tissue imaged on a Zeiss Airyscan microscope at 63× magnification, with the Z‐plane bisecting the widest part of a nucleus. The area of each nucleus in an image (> 30 nuclei in total for each age) was measured by manual outlining.
A1r eclipse ti microscope
Manufactured by Nikon
The Nikon A1R Eclipse Ti microscope is a high-performance research-grade instrument designed for advanced imaging applications. It features a modular design, allowing for customization to meet specific research needs. The microscope offers a wide range of advanced capabilities, including confocal imaging, high-speed acquisition, and multi-modal imaging capabilities.
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Lab products found in correlation
4 protocols using a1r eclipse ti microscope
1
Imaging and Quantifying Drosophila Fat Body
2
Quantifying Amyloid Plaque Burden
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The amyloid plaque load was measured in brain sagittal vibratome sections (60 μm) from mice transcardially perfused with PFA. The sections were stained for amyloid plaques using immunofluorescence with an Aβ primary antibody (6E10, against Aβ1-17, Sigma) after antigen retrieval in sodium citrate buffer. Antibody-antigen complexes were revealed using a DyLight 650conjugated goat anti mouse secondary antibody. DAPI (4′,6-diamidino-2-phenylindole) (Invitrogen) was used as counterstain. Digital images were taken on a Nikon A1R Eclipse Ti microscope.
3
Membrane Nanotube Force Measurements
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Tube pulling tension measurements were performed using a modified version of published setup15 (link), which allows simultaneous bright-field and FLIM imaging on an inverted Nikon Eclipse Ti A1R microscope. A membrane nanotube was formed by pulling away a micropipette-aspirated GUV, whose membrane was attached to a streptavidin-coated bead (3.05 mm diameter, Spherotec, Lake Forest, Illinois, USA) held in a fixed optical trap. The trap was obtained by focusing an ytterbium fiber laser (IPG laser, Burbach, Germany) through a 100X 1.3 NA oil immersion objective (Nikon, Tokyo, Japan). Force measurements were made measuring the displacement of the bead in the optical trap via a C-MOS Camera (Pixelink, Ottawa, Canada) with a home-made video recorder and bead tracking software under Matlab. The force F exerted on the bead was calculated from the Hooke’s law: F = k.Δx, where k is the stiffness of the trap (k = 8.58 pN.pix-1. W-1) and Δx the displacement of the bead from its initial, zero force position.
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Membrane Nanotube Force Measurements
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