Zeiss Axioplan 2 imaging and Axiophot 2 Guía de usuario

Zeiss Axioplan 2 imaging microscope and Axiovision software Microscopes 1 and 2 in room B501b User Guide Molecular Imaging Unit University of

7 Table 1. Microscope filter configurations. Microscope 1 Microscope 2 1 DAPI (Semrock 5060B) 1 DAPI (Semrock 5060B) 2 FITC (Chroma 41001) 2 FI

8 Figure 8. Buttons for changing filter position. Make sure that you are using a filter that matches the spectral properties of the fluorochromes

9 Microscope 2 Mag. Type Working distance (mm) Numerical Aperture (N.A.) Immersion medium Phase ring position DIC position 5x Plan-Neofl

10 5.2.2 Oil usage Only Zeiss F 158 immersion oil should be used with oil objectives. There are brown oil bottles on the table between the micros

11 Figure 11. Stage functions 7 Focusing There are focusing knobs on both sides of the microscope. The outer wheel is for coarse adjustment and

12 Figure 12. The button for overriding the focusing limit. 8 Contrasting methods for transmitted light imaging 8.1 Settings Each contrastin

13 Figure 13. Buttons for changing the condenser position (1) and an opening to view the current position (2). There are predefined settings

14 8.5 The auxiliary lens for transmitted light The button to control the auxiliary lens is right above the contrasting method indicator (number

15 9 Adjusting the oculars Make sure that the oculars are correctly adjusted for your eyes. If you have normal vision, the zero mark on the sides

16 Figure 16. Refresh rate settings for live image. 10.3 Acquiring a single image A single image can be acquired by clicking the snap-button (F

1 GENERAL ... 1 1.1...

17 Figure 18. Workarea menu Figure 19. Exposure settings 10.5 White balance White balance adjustment is not needed in fluorescence imagi

18 Figure 20. White balance settings 10.6 Image resolution Click Camera and then AxioCamHR in the Work area menu. Choose FRAME tab. There is a l

19 Figure 21. Display mapping adjustments 10.8 Acquiring multiple dyes Choose MULTIDIMENSIONAL ACQUISITION in the Work area menu and C

20 By clicking MEASURE (Figure 24) the software estimates the suitable time of exposure and the preview live window pops up (Figure 25). Adj

21 The AxioVision software is not able to use decimal numbers on the z slice thickness if the Windows user language is not set to English as it ca

22 Drive R: is your personal University network folder. You can access the same folder also from other Windows computers on the cam

23 in the image. If you have adjusted the display mapping curves (Chapter 10.7) you can save them in the exported image by selecting the APPLY DIS

24 On working days (Mo-Fr) leave the equipment on if your reservation ends before 16:00. If your reservation begins before 16:00 and ends after

25 There seems to be a way to avoid the problem: it occurs only when many image windows are open at the same time in cascade mode (windo

15.2 ... Computer 24 15.3 ...

1 1 General 1.1 Instrument This user guide covers the use of Zeiss Axioplan 2 imaging microscope. For more information on each instrument, visit

2 1.7 Sample preparation 1.7.1 Selection of fluorochromes If you want to achieve the best possible resolution, use a fluorochrome whose emission

3 Figure 1. Zeiss Axioplan microscope. 2.2 Light source for fluorescence imaging In case you want to use fluorescence light and it is not alr

4 2.3 Light source for transmitted light imaging The black round button labeled “HAL” for the halogen light source is on the right

5 Figure 4. Back side of the Zeiss Axiocam HRc camera 3 Light paths 3.1 Shutters for fluorescence light There are two shutters (Figure

6 3.2 Push rods There are two push rods near the oculars (Figure 6). The rod on the right (1) is for camera/ocular selection. The rod on the lef