[Histonet] query regarding silde preparation for LCM

[shazana hilda]

Dear Histonetters,

i've got few problem regarding the slide
preparation for laser microdissection. for your information, i'm using a PEN-membrane from PALM to be used on laser microdissection pressure catapult (LMPC), Ziess. According to the protocol i need to UV the slide prior
staining about 30min in order to make it hydrophilic. i've done it.
then continue fish the tissue after sectioned.(i'm using polyester wax
embedded similar like paraffin embedded jst this is low melting wax for
RNA work). then i've heat the slide on slide heater (to dewax) bout
45min at 37celcius. i've tried multiple period but seems like the
tissue hard to attach. after dip in 1st 100% ethanol (rehydration step) all tissue
detached. if using conventional immunohistochemistry protocol, the slide should be heat bout 1hr at 60celcius.i'm affraid if prolong heat the slide to 1hr at 37celcius will be detrimental to the RNA.
This slide will be stained suing Cresyl Violet.

Does anyone have any suggestion to solve this matter?

Many thanks.

Hilda

MSc.of Molecular Pathology,
Dept. of Pathology,
School of Medical Sciences,
Universiti Sains Malaysia,
16150 Kubang Kerian,
Kelantan, Malaysia.
Email: hilda_1075 <@t> yahoo.com