Glass bottom viewplate 96f

A blue fluorescence protein (BFP) cassette was amplified from the pFA6a-link-yomTagBFP2-Kan plasmid and integrated into the genome to generate an Hsp104-BFP fusion protein in all tested strains [77 (link)]. Log-phase cells growing in YPD at 25°C were transferred to a 37°C water bath and cells were collected at 0, 1, 2, and 3 hours after the temperature shift. The samples were resuspended in 200 μL PBS and transferred to a Glass Bottom ViewPlate-96F (PerkinElmer, Waltham, MA) that was precoated with concanavalin A (cat. no. C2010, Sigma-Aldrich). Images of Hsp104-BFP foci were acquired using an ImageXpress Micro XL system (Molecular Devices, San Jose, CA) and analyzed using a custom-built module under MetaXpress High-Content Image Acquisition and Analysis Software (Molecular Devices).

Yeast cells were diluted with PBS and loaded into a Glass Bottom ViewPlate®-96 F
(PerkinElmer, Waltham, MA) coated with concanavalin A (C2010, Sigma-Aldrich, St.
Louis, MO). Then the plates were centrifuged to attach cells to the glass bottom
and the images were obtained using the ImageXpress Micro XL system (Molecular
Device, Sunnyvale, CA). All the images were analyzed manually using ImageJ
(http://rsbweb.nih.gov/ij ).