User guide

1 GENERAL ......................................................................................................................... 1
1.1................................................................................................................... Instrument 1
1.2................................................................................................................. Reservations 1
1.3.......................................................................................................... Unauthorized use 1
1.4......................................................................................................... Users’ mailing list 1
1.5................................................................................................................... User guide 1
1.6..................................................................................................... Acknowledging MIU 1
1.7........................................................................................................ Sample preparation 2
1.7.1 .......................................................................................Selection of fluorochromes 2
1.7.2 .................................................................................................. Mounting samples 2
1.8......................................................................................... Image processing and analysis 2
2 TURNING ON THE EQUIPMENT ................................................................................ 2
2.1............................................................................... Zeiss Axioplan 2 imaging microscope 2
2.2............................................................................... Light source for fluorescence imaging 3
2.3.......................................................................... Light source for transmitted light imaging 4
2.4.................................................................................................... Axiocam HRc camera 4
3 LIGHT PATHS ................................................................................................................. 5
3.1..........................................................................................Shutters for fluorescence light 5
3.2..................................................................................................................... Push rods 6
4 FILTERS ........................................................................................................................... 6
5 OBJECTIVES ................................................................................................................... 8
5.1.............................................................................................. List of objectives available 8
5.2............................................................................................................... Oil objectives 9
5.2.1 ..................................................................... Changing between oil and air objectives 9
5.2.2 .............................................................................................................. Oil usage 10
5.2.3 ............................................................................................ Cleaning oil objectives 10
6 PLACING THE SAMPLE ............................................................................................. 10
7 FOCUSING ..................................................................................................................... 11
8 CONTRASTING METHODS FOR TRANSMITTED LIGHT IMAGING ............. 12
8.1....................................................................................................................... Settings 12
8.2...................................................................................................... Bright-field imaging 13
8.3................................................................................................... Phase contrast imaging 13
8.4................................................................................................................ DIC imaging 13
8.5.............................................................................. The auxiliary lens for transmitted light 14
8.6....................................... Optimal settings for transmitted light imaging (Köhler illumination) 14
9 ADJUSTING THE OCULARS...................................................................................... 15
10 ACQUIRING IMAGES .................................................................................................. 15
10.1 ............................................................................................. Zeiss Axiovision software 15
10.2 .................................................................................................................. Live view 15
10.3 .............................................................................................. Acquiring a single image 16
10.4 ............................................................................................................ Exposure time 16
10.5 ............................................................................................................. White balance 17
10.6 ......................................................................................................... Image resolution 18
10.7 ....................................................................................................... Display properties 18
10.8 ............................................................................................... Acquiring multiple dyes 19
10.9 .......................................................................................................... Z-stack imaging 20
11 POST PROCESSING ..................................................................................................... 21
11.1 ........................................................................................... Scalings and measurements 21
11.2 .......................................................................................................Shading correction 21
12 SAVING AND TRANSFERRING IMAGES ............................................................... 21
12.1 ............................................................................................................ Saving images 21
12.2 .................................................................................. Transferring images over network 22
12.2.1 ............................................................................................ Using MCBSERVER 22
12.2.2 ................................................................... Transferring directly to a Mac computer 22
13 EXPORTING IMAGES ................................................................................................. 22
14 ENDING YOUR IMAGING SESSION ........................................................................ 23
15 TROUBLESHOOTING ................................................................................................. 24
15.1 ............................................................................................................. Software help 24