KIF18B;EB1-GFP control and KO primary keratinocytes were isolated from mouse back skin as described above and plated on 3.5 mm glass-bottomed tissue culture (TC) plates (MatTek Corporation; P35G-1.5-14-C) coated with laminin. Calcium (0.5 mM) was added 4 h ahead of imaging to up-regulate cellular junctions and induce proliferation. Cells were imaged on an Andor XD Revolution Spinning Disk Confocal with temperature control chamber (37°C) and CO2 (5%) (Duke Light Microscopy Core Facility). Thirty-second movies (1 frame/second) were taken using a 60× Plan-Apo 1.2 NA water objective, Andor Ixon3 897 512 EMCCD camera, and MetaMorph Software. Postacquisition processing of movies was done with FIJI (ImageJ). EB1-GFP comets were tracked manually using the FIJI Manual Tracking Plugin. Ten EB1-GFP puncta were tracked per cell, five per spindle pole but across the entire astral region. The brightest puncta were selected for tracking.
Ixon3 897 512 emccd camera
Manufactured by Oxford Instruments
The Ixon3 897 512 EMCCD camera is a high-performance imaging device designed for scientific and industrial applications. It features a 512 x 512 pixel sensor and employs electron-multiplying charge-coupled device (EMCCD) technology to provide enhanced sensitivity and low-light imaging capabilities.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ixon3 897 512 emccd camera
1
Imaging Microtubule Dynamics in Keratinocytes
2
Tracking EB1-GFP comets in keratinocytes
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KIF18B;EB1-GFP control and KO primary keratinocytes were isolated from mouse back skin as described above and were plated on 3.5 mm glass-bottomed TC plates (MatTek Corporation, P35G-1.5-14-C) coated with laminin. 0.5 mM calcium was added 4 hours ahead of imaging to upregulate cellular junctions and induce proliferation. Cells were imaged on an Andor XD Revolution Spinning Disk Confocal with temperature control chamber (37 °C) and CO2 (5%) (Duke Light Microscopy Core Facility). 30 sec movies (1 frame/second) were taken using 60x Plan-Apo 1.2 NA water objective, Andor Ixon3 897 512 EMCCD camera and MetaMorph Software. Postacquisition processing of movies was done with FIJI (ImageJ). EB1-GFP comets were tracked manually using FIJI Manual Tracking Plugin. 10 EB1-GFP puncta were tracked per cell, 5 per spindle pole but across the entire astral region. The brightest puncta were selected for tracking.
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