Defining primary antibodies – Dako Autostainer User Manual

Page 54

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2. Select the desired reagent from the list using the UP and DOWN arrow keys on the

keyboard, the mouse pointer, the reagent list scroll bar or by typing the name of the
reagent. A dialog box prompts “Assign to the following unprogrammed slides?”.

Select YES to assign the selected reagent to the remaining slides in the grid and the next
programmable step is highlighted.

Select NO and the next programmable step is highlighted without assigning the reagent
to the remaining slides.

3. Select

the

NEXT button to complete the slide programming after all reagents are

programmed. The SLIDE LAYOUT MAP:PROGRAM SLIDES screen is displayed.

4. After

selecting

NEXT, the PROGRAMMING GRID will be checked for missing rinses. If

there is a rinse missing between two reagents, a message will ask if you want to continue.

Select YES to continue on the SLIDE LAYOUT MAP:PROGRAM SLIDES screen.

Select NO to return to the PROGRAMMING GRID and the missing rinse will appear on
the screen.

Defining Primary Antibodies

Each primary antibody reagent is programmed individually and will appear in the PROGRAMMING
GRID with its programmed pretreatment. (see Reagent Management, section 11) Primary
antibodies and/or pretreatments can be manual steps and are noted on the PROGRAMMING
GRID and printouts. If a reagent is a manual step, then that step is preformed before the slides
are loaded on the Autostainer (i.e. overnight incubations, Target Retrieval, etc.)

NOTE: A primary antibody’s pretreatment will automatically appear with the primary antibody, only
if the protocol template contains a pretreatment protocol element (see Designing a Protocol,
section 9).

1. Enter slide information and select a protocol template as described previously. (See

sections 8 and 9). The PROGRAMMING GRID is displayed listing the slides identified
during Slide Count or Slide Information entry. Add any detection reagents as described
above. When the first programmable Primary Antibody is highlighted (blinking), a list of
available Primary Antibodies for the step is displayed.

2. Select the desired antibody from the list using the UP and DOWN arrow keys on the

keyboard, the mouse pointer, the reagent list scroll bar or typing the name of the antibody
and press ENTER. The selected antibody and its pretreatment will appear on the
PROGRAMMING GRID.

NOTE: Pretreatments can be made a manual step (for more information on manual
steps, see Reagent Management, section 11) by positioning the mouse pointer over a
pretreatment, and pressing the right mouse button. To change the pretreatment back to a
step preformed on the Autostainer, position the mouse pointer over the pretreatment to
be changed and press the left mouse button. The list of available pretreatments is
displayed. Select the desired pretreatment from the list.

3. Negative controls are listed at the end of the primary antibody list and can be accessed

on the keyboard by pressing the SHIFT key and typing “_NC”. For example, for selecting
the negative control reagent for Vimentin, press the SHIFT key and type “_NCVimentin”.
Once the correct negative control is selected, press ENTER and the selected negative
reagent and pretreatment will appear on the PROGRAMMING GRID.