有关于免疫组化的问题
你可以按下面的方法试一试
PREPARATION AND STORAGE OF SPECIMENS:
SPECIMENS:
1.Frozen sections on adhesive-coated glass slides
2.Cell smears and cytocentrifuged cells (see procedure for the preparation of cytospins)
3.Fixative-fixed (Formalin, Bouin or B5), paraffin-embedded sections on adhesive-coated glass slides.
SAFETY NOTE: unfixed slides, cytospins, smears of human origin are to be considered potentially infectious. Therefore universal precautions in handling are mandatory.
FROZEN SECTIONS / SMEARS / CYTOCENTRIFUGED CELLS
Frozen sections / smears / cytospins are collected preferably, but not necessarily, on adhesive-coated slides (Fisher Superfrost Plus Cat. 12-550-15), and processed as follows within the next 48 hours:
1- air dry in a cool, dry place (AC) for 1hr up to 48 hr.
2- fix in absolute acetone (new aliquot) for 10 min at RT.
3- dry thoroughly for 15 min.
4- store at RT in a dry cool place (NOT the refrigerator, nor the freezer) for up to total of 48 hrs, from the time of preparation.
5- freeze.
To proceed with the immunostaining:
1- fix in 10% buffered formalin for 10 min at RT (NOTE rare antigens do not survive formalin fixation: proceed with the immunostains after acteone fixation).
2- wash thoroughly in PBS 3x
3- proceed with the immunostains without drying.
To store the frozen sections (see also here):
pair each slide with another glass slide (with or without section on) facing each other, with a short piece of thread in between. (NOTE you may put up to 5 pairs of slides in the same packet).
wrap in Saran Wrap tightly, so the slides are sealed.
label the slides legibly with a permanent marker
store at -20 o C (or better at -80 o C)
before open up the packet, allow the slides to reach room temperature.
