Adult fly guts were dissected in cold 1X PBS and fixed for 30 minutes in 1X PBS with 4% formaldehyde. Samples were washed three times in 1X PBS with 0.3% Triton X-100 and then mounted in Vectashield with DAPI (Vector Laboratories, H-1200). Confocal images were taken with Nikon Ti and Ti2 Spinning Disk at the Microscopy Resources. Adult fly phenotypes were imaged using a ZEISS Axiozoom V16 fluorescence microscope.
Ti2 spinning disk
Manufactured by Nikon
The Ti2 Spinning Disk is a laboratory equipment product from Nikon. It is a spinning disk confocal microscope system designed for live-cell imaging. The system utilizes dual Nipkow disks to provide high-speed optical sectioning and high-resolution imaging of fluorescent samples.
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Lab products found in correlation
2 protocols using ti2 spinning disk
1
Dissection and Imaging of Adult Fly Guts
2
Dual-species biofilm architecture visualization
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To comparatively visualize the architecture of single and dual-species biofilms, a cell suspension of 1 × 107 cells/mL of S. mutans was grown in RPMI alone or added to pre-adhered C. albicans in glass-bottom dishes (35 mm petri dish, MatTek Corporation). First, C. albicans (1 × 107 cells/mL) was incubated for 90 min at 37°C; and following incubation, dishes were washed with 10 mM PBS, and GFP-S. mutans was added, and the dishes were incubated statically at 37°C overnight. The following day, dishes were washed with 10 mM PBS and biofilms were stained with concanavalin-A-Alexa Fluor 647 (0.05 mg/mL) for 45 min at 37°C to stain for extracellular matrix. C. albicans hyphae were stained with 0.1% Calcofluor White, which stains cell wall chitin, for 10 min at room temperature. Samples were gently washed with 10 mM PBS and examined by confocal laser scanning microscopy at 40× and 60× magnifications (Nikon Ti2 Spinning Disk). Images were processed using Imaris and Photoshop CS6 software.
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