Michael Hooker Microscopy Facility (MHMF.ORG)

How To's

Acquisition:

• Background Correction of unevenness of illumination for transmitted light images

Viewing:

• Zeiss 510 confocal image databases (LSM Image Browser, Windows)
• Leica SP2 confocal image databases (LAS AF lite v2.6.3 + software - Windows XP/7/8  or  LCS Lite software - WindowsXP only  )
• Olympus FV1000 image databases FV10-ASW_Viewer3_1.zip (Windows)  (FV10-ASW 2.0c Viewer)
• ImageJ - Short Cut Keys (prn)
• Volocity on the  \\CCD,  \\Schott  or  \\Nomarski workstations  or  your computer (Views Leica, Zeiss & Olympus confocal image databases)

Processing:

• General Image overlaying
• Convert B/W image to color using Photoshop
• Overlaying red/green images using Photoshop
• Image registration / realignment

• Processing images: Calculate area of bright cells vs the background

Analysis:

• Cell / particle counting with C-Imaging
• Measuring black background area using ImageJ
• Measuring percent of area of bright cells versus the background area using C-Imaging

Movies:

• Making movies from Leica SP2 tiff image series (z-sections or time lapse)
• Digitizing VHS tapes on the computer called \\Young

Technical Tasks:

• Adding Scale bars: Keeping records  |  Leica confocal images  |  Zeiss confocal images  |  Spinning disk confocal    widefield scopes  |  C-Imaging  |  Metamorph  |  Image-J  |  Photoshop  |  tricks  |  traps
• Excel - Making a frequency Distribution Histogram
• Measuring fraction of area taken up by a labeled structure (e.g. DAB)
• http://www.luminous-landscape.com/tutorials/digital-blending.shtml 

		Copyright 2001-2014  Dr. M. Chua, Program in Molecular Biology & Biotechnology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599
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