Olympus Fluoview-1000 - Page 3

.........................................................................................................................................Bleedthrough. 33
......................................................................................................................Reducing bleedthrough 33
.................................................................................................Procedure to minimize bleedthrough 33
.....................................................................................................................................................Noise. 34
................................................................................................................Signal Noise (Shot Noise). 34
..................................................................................................................................Detector Noise. 35
............................................................................................................................Kalman Averaging. 35
............................................................................................................................................Background 35
.....................................................................................Bright Images versus Scorching the Specimen 35
.....................................................................................................................Images with Weak Signals 35
...........................................................................................................................................Using Zoom 36
..........................................................................................Zoom reduces objective lens aberrations. 36
..........................................................Use Zoom to fulfill sampling requirements for the specimen. 36
.......................................................................Use Zoom to fulfill adequately sample the objective. 36
.............................................................................................................................Obtaining a Z-Series. 37
.............................................................................................................................................Resolution. 37
................................................................................Sampling frequency and the Nyquist Theorem. 38
............................................................................................................................................Box size. 38
Zoom. 38
..........................................................................................Considerations for increasing resolution. 38
.........................................................................................................................................Size of Pixels 39
..........................................Table. 5 Pixel Size (µm) versus Box Size and Magnification at Zoom = 1 39
................................................................................................................................Microscope controls. 41
.....................................................................................................................................................Focus. 41
............................................................................................................................Fine/Coarse Focus. 41
...................................................................................................................Lowering the Objectives. 41
..................................................................................................................................Manual shutter. 41
..............................................................................................Differential Interference Contrast (DIC). 41
....................................................................................................................................DIC Analyzer. 41
....................................................................................................................................DIC polarizer. 42
........................................................................................................................................DIC mirror. 42
................................................................................................................Setting Koehler Illumination. 42
...........................................................................................................................Condenser position. 43
........................................................................................................................Photoactivation Methods. 44
.....................................................................................................................Zoom versus Laser Power. 44
...........................................................................................................Recording initial recovery event. 44
..........................................................................................................Methods to photoactivate/bleach. 44
.............................................................................................................Using the Stimulus Window. 44
..................................................................................................Avoid spontaneous photoactivation. 46
...........................................................................................Controlling Computer Display Brightness. 46
..................................................................................................................Set ‘Darkroom’ color mode. 47
.............................................................................................................................Technical Information 48
.............................................................................................................................Caring for Objectives 48
Olympus Fluoview-1000 User’s Guide
V.M. Bloedel Hearing Research Center, Core for Communication Research
Center on Human Development and Disability, Digital Microscopy Center
May 11, 2011 3