Shreffler: Hoerscht Counterstaining

Equipment

 * Frosted glass slides, pre-cleaned (Cat. 12-550-343; Fisherbrand)
 * Cytospin machine
 * Microscope
 * Coplin jar

Reagents

 * Methanol (Cat. 494437-1L; Sigma-Aldrich)
 * Hoescht dye aliquots acquired from Mempel lab (unsure if Hoechst 33258 or Hoechst 33342)

Procedure

 * 1) Cytospin cells onto a frosted slide, pre-cleaned (600 rpm, 5 min, approx. 500 µl cell suspension used).
 * 2) Dry for 15 minutes at RT.
 * 3) Place in 100% methanol for 1 min.
 * 4) Remove and dry for 5 min at RT (keep in dark).
 * 5) Meanwhile, mix Hoescht dye by adding 5 µl of aliquoted stock to 50 ml of diH20 in a Coplin jar.
 * 6) Place slide in the in the Hoescht staining solution in the Coplin jar for 1 min.
 * 7) Wash in diH20 for 5 min without agitation.
 * 8) Dry at RT in the dark for at least 15 min.
 * 9) Analyze by microscopy.