Imaging Resource

Standard Operating Procedure

CYTOGENETIC WORK STATION

PROCEDURE TITLE

Capturing images on the Zeiss Axioplan 2 microscope using Isis.

PROCEDURE

1Switch on the microscope using the green rocker switch at the back right hand side of the microscope.

2Switch on the UV power supply if required. Green rocker switch on the Ebq 100-supply, which is located on the left hand side of the microscope.

3Switch on the computer and log on as normal,make sure workstation only is not selected.

4Press the HAL button on the right side of the microscope to put on or off the halogen lamp, which is used for bright field images.

5The filter wheel, which contains a complete set of filters for M-FISH, is situated directly under the eyepieces. The number tells you which filter is in place.

  1. 1=DAPI.
  2. 2=AQUA.
  3. 3=FITC.
  4. 4=SPO.
  5. 5=TRITC
  6. 6=CY5
  7. 7+8=BLANK

Pressing either of the black buttons under the focus wheel on the left hand side of the microscope will move the filter position

6The stage can be moved manually by using the two silver knobs at the front and on the left side of

The stage.

7The slide can be put in place and then the desired objective can be selected by moving the turret by

hand or by pressing either of the black buttons under the focus knob on the right side of the microscope.

8Switch on the light required HAL button (white light) orFL button (fluorescence).

9Focus on the sample

10Double click on the Isis icon to start the programme.

11Under users login select.

  1. User=adm
  2. Root path=data
  3. Case name=Enter the name of the sample and select OK.

12Select the experiment type from the list in the lower right corner of the screen. Eg;BGR=blue green

Red. Select Data directory right corner above the experiment type. ( 4 letters max )

13To capture all the color channels sequentially click on the spectrum symbol in the upper right corner

Of the screen.

13Change the light path to the camera | push in black knob on the left side of the eyepieces.

14Move the mouse pointer to the capture control and select by left mouse clicking.

15Focus on the image.

16Left mouse click to start capture.

17Right mouse click to terminate capture sequence Esc key to cancel if capture image is no good.

18Select file save. The image will be saved to your network area underN: images/msd as a MCI image file. Always save a file in this format so you can reopen the image in ISIS.

19To save an overlay as a TIFF file press the X key on the keyboard and save as a TIFF to your network area. Do not apply color modifications.

20To save a single color from the overlay select | individual color so only one color is displayed on the

screen and then press the X key on the keyboard as before to save as a TIFF. This is a 8bit Tiff image.

21Repeat process for all images.

22Open the Tiff file in Photoshop and resave.

23The file will now open in Powerpoint.

Imaging FacilityPage 111/15/2018